Browsing by Author "Vipin Rai"

Now showing 1 - 20 of 25

A viscochromic, mechanochromic, and unsymmetrical azine for selective detection of Al3+ and Cu2+ ions and its mitotracking studies
(Royal Society of Chemistry, 2019) Richa Yadav; Abhishek Rai; Avinash Kumar Sonkar; Vipin Rai; Subash Chandra Gupta; Lallan Mishra
A hydrazone obtained by the reaction of 2-hydroxynaphthaldehyde with hydrazine hydrate was allowed to react further with 4-diethylamino-2-hdroxybenzaldehyde to provide an unsymmetrical azine derivative (NDEA). The azine has been characterized using spectroscopic techniques and single crystal X-ray crystallography. It selectively detects Al3+ and Cu2+ ions in aqueous methanol (DMSO:H2O:MeOH = 0.1:1.9:8.0, v/v, HEPES buffer, pH 7.4) and exhibits naked eye visible color changes from greenish yellow to bright yellow and brown on the addition of Al3+ and Cu2+ ions, respectively, with concomitant changes in their absorption spectra. The corresponding solutions containing Al3+ and Cu2+ ions also display changes in their emission spectra via "TURN ON" and "TURN OFF" pathways with a detection limit of 1.65 × 10-7 M and 1.52 × 10-7 M, respectively. NDEA works as a reversible probe towards Al3+ ions and Cu2+ ions by the addition of F- ions and EDTA2- ions, respectively. The complexes of NDEA, separately with Al3+ and Cu2+ ions, have been optimized by DFT (density functional theory). It also exhibits viscochromic and mechanochromic properties. Most importantly, NDEA significantly detects these ions in rat C6 glioma cell lines and displays good cell permeability. It also co-localizes with the commercially available Mitotracker Red and exhibits a unique application as a live cell mitochondrial tracker. Practically, NDEA detects Al3+ and Cu2+ ions in real water samples and blood serum with a high accuracy. © The Royal Society of Chemistry and the Centre National de la Recherche Scientifique.
Anti-cancer activities of Bharangin against breast cancer: Evidence for the role of NF-κB and lncRNAs
(Elsevier B.V., 2018) Nikee Awasthee; Vipin Rai; Sumit S. Verma; K. Sajin Francis; Mangalam S. Nair; Subash C. Gupta
Breast cancer remains one of the leading causes of cancer related deaths in women worldwide. Bharangin is a diterpenoid quinonemethide that has demonstrated therapeutic potential against leukemia, lymphoma, and multiple myeloma cells. Whether this diterpenoid exhibit activities against breast cancer cells and the underlying mechanism is largely unknown. Herein, we provide evidence that bharangin suppresses the proliferation of MCF-7, MDA-MB-231, MDA-MB-453, MDA-MB-468 and T-47D breast cancer cells. As examined by AO/PI staining, DAPI staining, sub-G1 analysis, phosphatidylserine externalization, caspase activation, DNA laddering, and poly-ADP ribose polymerase cleavage, the diterpenoid induced apoptosis in breast cancer cells. The growth inhibitory effect of bharangin on breast cancer cells was further confirmed from colony-formation assay. Furthermore, the cancer cell migration was also suppressed by the diterpenoid. Mechanistically, bharangin was found to modulate multiple cancer related cell signalling pathways in breast cancer cells. Bharangin suppressed the expression of cell survival and invasive proteins, and induced Bax and mitochondrial depolarization in breast cancer cells. The diterpenoid also suppressed the activation of pro-inflammatory transcription factor, nuclear factor (NF)-κB induced by okadaic acid. Finally, the diterpenoid induced the expression of tumor suppressor lncRNAs (MEG-3, GAS-5), while down-regulating oncogenic H19 expression. Overall, these results suggest that bharangin exhibits anti-carcinogenic, anti-proliferative and anti-inflammatory activities against breast cancer cells. The modulation of lncRNA expression and inhibition of NF-κB activation by bharangin may contribute to its anti-carcinogenic activities. © 2018 Elsevier B.V.
Clinical significance of long noncoding RNAs in breast cancer patients
(Elsevier, 2022) Nikee Awasthee; Anusmita Shekher; Vipin Rai; Pranjal K. Baruah; Anurag Sharma; Kishore B. Challagundla; Subash C. Gupta
Breast cancer is one of the commonly diagnosed cancers in women globally. The long noncoding RNAs (lncRNAs) belong to the category of noncoding RNAs. The lncRNAs are equal to or more than 200 nucleotides in length. Accumulating evidence over the last 5 years has indicated that lncRNAs regulate crucial steps of tumorigenesis in multiple cancer types. Because of high dysregulation, specificity, and sensitivity in the detection, lncRNAs possess the potential of diagnostic and prognostic marker for breast cancer. The commonly identified lncRNAs in breast cancer include antisense ncRNA in the INK4 locus, breast cancer antiestrogen resistance 4, differentiation antagonizing nonprotein-coding RNA, H19-imprinted maternally expressed transcript, hypoxia-inducible factor 1A-antisense RNA 2, HOX transcript antisense RNA, metastasis-associated lung adenocarcinoma transcript 1, miR-503 host gene, terminal differentiation-induced ncRNA, TROJAN, and urothelial cancer-associated 1. The most common techniques used for elucidating the functions of lncRNAs in breast cancer patients include reverse transcription-quantitative polymerase chain reaction, next-generation sequencing, RNA-sequencing, microarray, TILING array, serial analysis of gene expression, cap analysis gene expression, and computational methods. The clinical specimens used to detect lncRNAs in patients include serum, plasma, tissues, urine, and saliva. The lncRNAs are reported to modulate multiple cell signaling pathways in breast cancer patients. The dysregulation in lncRNA expression is also one of the mechanisms by which breast cancer cells develop resistance to therapeutic agents. Thus breast cancer cells could also be sensitized to the therapeutic agents by targeting lncRNAs. The common strategies for targeting lncRNAs include the use of small molecules, antisense oligonucleotides, and RNA interference (RNAi) technology. The focus of this chapter is to review the potential of lncRNAs in breast cancer patients. © 2022 Elsevier Inc. All rights reserved.
Curcuma raktakanda induces apoptosis and suppresses migration in cancer cells: Role of reactive oxygen species
(MDPI AG, 2019) Shruti Mishra; Sumit Singh Verma; Vipin Rai; Nikee Awasthee; Jayadev S. Arya; Kaustabh K. Maiti; Subash C. Gupta
Although over 100 species of Curcuma are reported, only Curcuma longa is extensively studied. Curcuma raktakanda, a poorly studied species, is most commonly distributed in the Kerala state of India. For the first time, we examined the efficacy of different fractions (acetone, hexane, and ethyl acetate) of C. raktakanda against glioma, cervical, and breast cancer cell lines. As determined by mitochondrial reductase activity assay, the viability of cancer cells was decreased in a concentration-dependent manner by the three fractions. The half maximal inhibitory concentration (IC-50) values after the treatment of C-6 glioma cells for 48 h was found to be 32.97 μg/mL (acetone extract), 40.63 μg/mL (hexane extract), and 51.65 μg/mL (ethyl acetate extract). Of the three fractions, the acetone fraction was more effective. The long-term colony formation of cancer cells was significantly suppressed by the acetone fraction. Analyses using DAPI (4′,6- diamidino-2-phenylindole) staining, AO/PI (acridine orange/propidium iodide) staining, DNA laddering, and sub-G1 population revealed that the acetone extract induced apoptosis in glioma cells. The extract induced reactive oxygen species generation and suppressed the expression of cell survival proteins. The migration of cancer cells was also suppressed by the acetone extract. The gas chromatography-mass spectrometry (GC-MS) analysis indicated that tetracontane, dotriacontane, hexatriacontane, pentacosane, hexacosane, and eicosane are the major components in the acetone extract. Collectively, the extract from C. raktakanda exhibited anti-carcinogenic activities in cancer cells. We are exploring whether the phytoconstituents, individually, or collectively contribute to the anti-cancer activities of C. raktakanda. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
Efficacy of Cannabis and its Constituents in Disease Management: Insights from Clinical Studies
(Bentham Science Publishers, 2023) Divya Vimal; Leonard Clinton D’souza; Vipin Rai; Samridhi Lal; Anurag Sharma; Subash Chandra Gupta
There is a long history of informal use of Cannabis sativa (commonly called cannabis) for many purposes, including treating various ailments worldwide. However, the legalization of cannabis in multiple countries, specifically for medical purposes, has grabbed the researchers' attention to discover the scientific evidence regarding cannabis’s beneficial effects. Among over 500 identified compounds (cannabinoids), Δ9-Tetrahydro-cannabinol (THC) and cannabidiol (CBD) are two major active cannabinoids derived from cannabis. Cannabinoids exert their effects through cannabinoid receptors (CB1R and CB2R). In the recent past, clinical trials have shown the efficacy of cannabis and cannabinoids for various human ailments, such as cancer, neurological disorders, inflammatory bowel disease, chronic pain, and metabolic disorders. The commonly used constituents and derivatives of cannabis include CBD, THC, THCV, dronabinol, nabilone, and nabiximol. The cannabis constituents have also been used in combination with other agents, such as megestrol acetate, in some clinical trials. The common routes for the administration of cannabis are oral, sublingual, or topical. Cannabis has also been con-sumed through smoking, inhalation, or with food and tea. A maximum of 572 patients and a minimum of nine patients have participated in a single clinical trial. Cannabis is le-galized in some countries with restrictions, such as Belize, Canada, Colombia, Costa Ri-ca, The Czech Republic, Jamaica, Netherlands, South Africa, Spain, and Uruguay. This article provides a compilation of published studies focusing on clinal trials on the therapeutic effects of cannabis. The adverse effects of cannabis and its constituents are also discussed. © 2023 Bentham Science Publishers.
Epoxyazadiradione exhibit activities in head and neck squamous cell carcinoma by targeting multiple pathways
(Springer, 2020) Vipin Rai; Sushil Kumar Aggarwal; Sumit Singh Verma; Nikee Awasthee; Anupam Dhasmana; Sadhna Aggarwal; Satya N. Das; Mangalam S. Nair; Sanjay Yadav; Subash C. Gupta
The head and neck squamous cell carcinoma (HNSCC) constitute about 90% of all head and neck cancers. HNSCC falls in the top 10 cancers in men globally. Epoxyazadiradione (EPA) and Azadiradione (AZA) are the limonoids derived from the medicinal plant Azadirachta indica (popularly known as Neem). Whether or not the limonoids exhibit activities against HNSCC and the associated mechanism remains elusive. Herein, we demonstrate that EPA exhibits stronger activity in HNSCC in comparison to AZA. The limonoids obeyed the Lipinski’s rule of 5. EPA exhibited activities in a variety of HNSCC lines like suppression of the proliferation and the induction of apoptosis. The limonoid suppressed the level of proteins associated with anti-apoptosis (survivin, Bcl-2, Bcl-xL), proliferation (cyclin D1), and invasion (MMP-9). Further, the expression of proapoptotic Bax and caspase-9 cleavage was induced by the limonoid. Exposure of EPA induced reactive oxygen species (ROS) generation in the FaDu cells. N-acetyl-L-cysteine (ROS scavenger) abrogated the down-regulation of tumorigenic proteins caused by EPA exposure. EPA induced NOX-5 while suppressing the expression of programmed death-ligand 1 (PD-L1). Further, hydrogen peroxide induced NF-κB-p65 nuclear translocation and EPA inhibited the translocation. Finally, EPA modulated the expression of lncRNAs in HNSCC lines. Overall, these results have shown that EPA exhibit activities against HNSCC by targeting multiple cancer related signalling molecules. Currently, we are evaluating the efficacy of this molecule in mice models. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.
Evaluation of antioxidant, anti-inflammatory and anticancer activities of diosgenin enriched Paris polyphylla rhizome extract of Indian Himalayan landraces
(Elsevier Ireland Ltd, 2021) Debmalya Das Gupta; Shruti Mishra; Sumit Singh Verma; Anusmita Shekher; Vipin Rai; Nikee Awasthee; Tridip J. Das; Dipayan Paul; Sanjib K. Das; Hui Tag; Subash Chandra Gupta; Pallabi K. Hui
Ethnopharmacological relevance: Traditional medicinal plants have gained attention as a potential therapeutic agent to combat cancer and inflammation. Diosgenin rich fresh extracts of Paris polyphylla rhizome from Indian Himalaya is traditionally used as wound healing, anti-bleeding, anti-inflammatory and anti-cancer agent by the folk healers. Aim of the study: Present study was aimed to prepare two types of extracts from Paris polyphylla rhizome of Indian Himalayan landraces – 1. ethanolic extract of Paris polyphylla rhizome (EEPPR) and 2. Diosgenin enriched Paris polyphylla rhizome extract (DPPE), quantification of diosgenin content, and to evaluate their in vitro anti-oxidant, in vivo anti-inflammatory and in vitro cytotoxicity and anti-cancer activities of the DPPE. Materials and methods: Diosgenin content of EEPPR was quantified through GC-MS while diosgenin content of DPPE was quantified through HPTLC, and the diosgenin yield from EEPPR and DPPE were compared. In vitro antioxidant activities of DPPE were performed using DPPH, NOD, RP and SOD assay while in vivo anti-inflammatory activity of DPPE were evaluated in dextran induced hind paw edema in rats. In vitro cytotoxicity and anti-cancer activities of DPPE were evaluated in human breast cancer cell lines (MCF-7, MDA-MB-231), cervical cancer cell lines (HeLa) and Hep-2 cell lines. Results: EEPPR obtained through cold extraction method using 70% ethanol showed maximum diosgenin content of 17.90% quantified through GC-MS while similar compounds pennogenin (3.29%), 7β-Dehydrodiosgenin (1.90%), 7-Ketodiosgenin acetate (1.14%), and 7 β-hydroxydiosgenin (0.55%) were detected in low concentration, and thus confirmed diosgenin as major and lead phytochemical. However, DPPE obtained through both cold and repeated hot extraction with the same solvent (70% ethanol) showed diosgenin content of 60.29% which is significantly higher (p < 0.001) than the diosgenin content in EEPPR. DPPE demonstrated significant in vitro antioxidant activities by dose-dependently quenched (p < 0.001) SOD free radicals by 76.66%, followed by DPPH (71.43%), NOD (67.35%), and RP (63.74%) at a max concentration of 2 μg/μl of ascorbic acid and test drugs with remarkable IC50 values (p < 0.01). Further, DPPE also showed potent anti-inflammatory activities by dose-dependently suppressed dextran induced paw edema in rats (p < 0.01) from 2 h to 4 h. DPPE suppressed the proliferation of MCF-7, MDA-MB-231, Hep-2 and HeLa cell lines. Maximum activity was observed in MCF-7 cells. The DPPE also induced apoptosis in MCF-7 cell lines as measured by AO/PI and DAPI staining, as well as DNA laddering, cell cycle analysis and phosphatidylserine externalization assay. The growth-inhibitory effect of DPPE on MCF-7 breast cancer cells was further confirmed from the colony-formation assay. DPPE upregulated expression of Bax and downregulated Bcl-2 and survivin mRNA transcripts. Conclusion: DPPE obtained through both cold and repeated hot extraction using ethanol showed significantly higher content of diosgenin than the diosgenin content detected in EEPPR. However, diosgenin yield of both the extracts (EEPPR & DPPE) clearly confirmed diosgenin as major and lead phytochemical of Paris polyphylla rhizome of Indian Himalayan landraces. Further, DPPE also demonstrated potent in vitro anti-oxidative and in vivo anti-inflammatory activities and showed in vitro cytotoxicity and significant anti-cancer (apoptosis) effects in MCF-7 breast cancer cells. © 2021
Flavonoids as Promising Neuroprotectants and Their Therapeutic Potential against Alzheimer's Disease
(Hindawi Limited, 2022) Tarun Minocha; Hareram Birla; Ahmad A. Obaid; Vipin Rai; P. Sushma; Chandan Shivamallu; Mahmoud Moustafa; Mohammed Al-Shehri; Ahmed Al-Emam; Maria A. Tikhonova; Sanjeev Kumar Yadav; Burkhard Poeggeler; Divakar Singh; Sandeep Kumar Singh
Alzheimer's disease (AD) is one of the serious and progressive neurodegenerative disorders in the elderly worldwide. Various genetic, environmental, and lifestyle factors are associated with its pathogenesis that affect neuronal cells to degenerate over the period of time. AD is characterized by cognitive dysfunctions, behavioural disability, and psychological impairments due to the accumulation of amyloid beta (Aβ) peptides and neurofibrillary tangles (NFT). Several research reports have shown that flavonoids are the polyphenolic compounds that significantly improve cognitive functions and inhibit or delay the amyloid beta aggregation or NFT formation in AD. Current research has uncovered that dietary use of flavonoid-rich food sources essentially increases intellectual abilities and postpones or hinders the senescence cycle and related neurodegenerative problems including AD. During AD pathogenesis, multiple signalling pathways are involved and to target a single pathway may relieve the symptoms but not provides the permanent cure. Flavonoids communicate with different signalling pathways and adjust their activities, accordingly prompting valuable neuroprotective impacts. Flavonoids likewise hamper the movement of obsessive indications of neurodegenerative disorders by hindering neuronal apoptosis incited by neurotoxic substances. In this short review, we briefly discussed about the classification of flavonoids and their neuroprotective properties that could be used as a potential source for the treatment of AD. In this review, we also highlight the structural features of flavonoids, their beneficial roles in human health, and significance in plants as well as their microbial production. © 2022 Tarun Minocha et al.
Health benefits of resveratrol: Evidence from clinical studies
(John Wiley and Sons Inc, 2019) Akhand Pratap Singh; Rachna Singh; Sumit Singh Verma; Vipin Rai; Catherine H. Kaschula; Pralay Maiti; Subash Chandra Gupta
Resveratrol is a polyphenolic nutraceutical that exhibits pleiotropic activities in human subjects. The efficacy, safety, and pharmacokinetics of resveratrol have been documented in over 244 clinical trials, with an additional 27 clinical trials currently ongoing. Resveretrol is reported to potentially improve the therapeutic outcome in patients suffering from diabetes mellitus, obesity, colorectal cancer, breast cancer, multiple myeloma, metabolic syndrome, hypertension, Alzheimer's disease, stroke, cardiovascular diseases, kidney diseases, inflammatory diseases, and rhinopharyngitis. The polyphenol is reported to be safe at doses up to 5 g/d, when used either alone or as a combination therapy. The molecular basis for the pleiotropic activities of resveratrol are based on its ability to modulate multiple cell signaling molecules such as cytokines, caspases, matrix metalloproteinases, Wnt, nuclear factor-κB, Notch, 5′-AMP-activated protein kinase, intercellular adhesion molecule, vascular cell adhesion molecule, sirtuin type 1, peroxisome proliferator-activated receptor-γ coactivator 1α, insulin-like growth factor 1, insulin-like growth factor-binding protein 3, Ras association domain family 1α, pAkt, vascular endothelial growth factor, cyclooxygenase 2, nuclear factor erythroid 2 like 2, and Kelch-like ECH–associated protein 1. Although the clinical utility of resveratrol is well documented, the rapid metabolism and poor bioavailability have limited its therapeutic use. In this regard, the recently produced micronized resveratrol formulation called SRT501, shows promise. This review discusses the currently available clinical data on resveratrol in the prevention, management, and treatment of various diseases and disorders. Based on the current evidence, the potential utility of this molecule in the clinic is discussed. © 2019 Wiley Periodicals, Inc.
Isodeoxyelephantopin, a Sesquiterpene Lactone Induces ROS Generation, Suppresses NF-κB Activation, Modulates LncRNA Expression and Exhibit Activities Against Breast Cancer
(Nature Research, 2019) Sumit S. Verma; Vipin Rai; Nikee Awasthee; Anupam Dhasmana; Dhanya S Rajalaksmi; Mangalam S. Nair; Subash C. Gupta
The sesquiterpene lactones, Isodeoxyelephantopin (IDET) and Deoxyelephantopin (DET) are known to exhibit activities against some cancer types. The activities of these lactones against breast cancer and the molecular bases is not known. We examined the efficacy of lactones in breast cancer preclinical model. Although both lactones exhibited drug like properties, IDET was relatively effective in comparison to DET. IDET suppressed the proliferation of both invasive and non-invasive breast cancer cell lines. IDET also suppressed the colony formation and migration of breast cancer cells. The assays for Acridine Orange (AO)/Propidium Iodide (PI) staining, cell cycle distribution, phosphatidylserine externalization and DNA laddering suggested the apoptosis inducing potential of IDET. The treatment with IDET also induced an accumulation of cells in the sub-G1 and G2/M phases. The exposure of breast cancer cells to the lactone was associated with a depolarization in mitochondrial membrane potential, and cleavage of caspase and PARP. The lactone induced reactive oxygen species (ROS) generation in breast cancer cells. Further, the use of N-acetyl cysteine (NAC) suppressed IDET induced ROS generation and apoptosis. The NF-κB-p65 nuclear translocation induced by okadaic acid (OA) was suppressed by the sesquiterpene. IDET also suppressed the expression of NF-κB regulated tumorigenic proteins, and induced the expression of proapoptotic gene (Bax) in cancer cells. While the expression of oncogenic lncRNAs was suppressed, the tumor suppressor lncRNAs were induced by the sesquiterpene. Collectively, the modulation of multiple cell signaling molecules by IDET may contribute to its activities in breast cancer cells. © 2019, The Author(s).
Long non-coding RNAs and nuclear factor-κB crosstalk in cancer and other human diseases
(Elsevier B.V., 2020) Subash C. Gupta; Nikee Awasthee; Vipin Rai; Srinivas Chava; Venugopal Gunda; Kishore B. Challagundla
The regulation of the pleiotropic transcription factor, nuclear factor-κB (NF-κB) by miRNAs and proteins is extensively studied. More recently, the NF-κB signaling was also reported to be regulated by several long non-coding RNAs (lncRNAs) that constitute the major portion of the noncoding component of the human genome. The common NF-κB associated lncRNAs include NKILA, HOTAIR, MALAT1, ANRIL, Lethe, MIR31HG, and PACER. The lncRNA and NF-κB signaling crosstalk during cancer and other diseases such as cardiomyopathy, celiac disease, cerebral infarction, chronic kidney disease, diabetes mellitus, Kawasaki disease, pregnancy loss, and rheumatoid arthritis. Some NF-κB related lncRNAs can affect gene expression without modulating NF-κB signaling. Most of the lncRNAs with a potential to modulate NF-κB signaling are regulated by NF-κB itself suggesting a feedback regulation. The discovery of lncRNAs have provided a new type of regulation for the NF-κB signaling and thus could be explored for therapeutic interventions. The manner in which lncRNA and NF-κB crosstalk affects human pathophysiology is discussed in this review. The challenges associated with the therapeutic interventions of this crosstalk are also discussed. © 2019 Elsevier B.V.
Long non-coding RNAs are emerging targets of phytochemicals for cancer and other chronic diseases
(Springer Science and Business Media Deutschland GmbH, 2019) Shruti Mishra; Sumit S. Verma; Vipin Rai; Nikee Awasthee; Srinivas Chava; Kam Man Hui; Alan Prem Kumar; Kishore B. Challagundla; Gautam Sethi; Subash C. Gupta
The long non-coding RNAs (lncRNAs) are the crucial regulators of human chronic diseases. Therefore, approaches such as antisense oligonucleotides, RNAi technology, and small molecule inhibitors have been used for the therapeutic targeting of lncRNAs. During the last decade, phytochemicals and nutraceuticals have been explored for their potential against lncRNAs. The common lncRNAs known to be modulated by phytochemicals include ROR, PVT1, HOTAIR, MALAT1, H19, MEG3, PCAT29, PANDAR, NEAT1, and GAS5. The phytochemicals such as curcumin, resveratrol, sulforaphane, berberine, EGCG, and gambogic acid have been examined against lncRNAs. In some cases, formulation of phytochemicals has also been used. The disease models where phytochemicals have been demonstrated to modulate lncRNAs expression include cancer, rheumatoid arthritis, osteoarthritis, and nonalcoholic fatty liver disease. The regulation of lncRNAs by phytochemicals can affect multi-steps of tumor development. When administered in combination with the conventional drugs, phytochemicals can also produce synergistic effects on lncRNAs leading to the sensitization of cancer cells. Phytochemicals target lncRNAs either directly or indirectly by affecting a wide variety of upstream molecules. However, the potential of phytochemicals against lncRNAs has been demonstrated mostly by preclinical studies in cancer models. How the modulation of lncRNAs by phytochemicals produce therapeutic effects on cancer and other chronic diseases is discussed in this review. © Springer Nature Switzerland AG 2019.
Melatonin induces apoptosis and cell cycle arrest in cervical cancer cells via inhibition of NF-κB pathway
(Springer Science and Business Media Deutschland GmbH, 2022) Tarun Minocha; Megha Das; Vipin Rai; Sumit Singh Verma; Nikee Awasthee; Subash Chandra Gupta; Chandana Haldar; Sanjeev Kumar Yadav
Cervical cancer is the most prevalent cancer in females. Melatonin, a neurohormone has been documented as a promising therapeutic molecule for cervical cancer. However, the underlying molecular mechanism is not known. We explored the dose-dependent anti-tumor response of melatonin against cervical cancer cell lines, HeLa (HPV-18 positive) and SiHa (HPV-16 positive). The anti-cancer effect of melatonin was evaluated by MTT assay, cell imaging, colony formation, DAPI, AO/PI, LDH, Flow cytometry, scratch assay, western blot analysis and real-time PCR. Results of DAPI, AO/PI, LDH, and Annexin/PI staining revealed that melatonin induces apoptosis. The results of cell cycle analysis revealed that melatonin arrests the HeLa and SiHa cells in sub-G1 and G1 phases, respectively. Western blot analysis revealed that melatonin downregulated the expression of pro-inflammatory transcription factor, NF-κB and the expression of COX-2 protein, a key mediator in cell proliferation. In addition, melatonin downregulated the expression of an invasive marker, MMP-9, an antiapoptotic protein, Bcl-2, and upregulated the expression of pro-apoptotic protein, Bax at both transcriptional and translational levels. Overall, the results suggest that melatonin exhibited strong anti-cancer therapeutic potential against human cervical cancer cell line progression possibly through inhibition of NF-κB signalling pathway. © 2022, The Author(s), under exclusive licence to Springer Nature Switzerland AG.
Molecular Diagnostics in Pancreatic Cancer
(Springer Singapore, 2019) Shruti Mishra; Vipin Rai; Abhai Kumar; Sushil Kumar Aggarwal; Subash Chandra Gupta
[No abstract available]
Moringin, an isothiocyanate modulates multiple cellular signalling molecules in breast cancer cells
(Elsevier Inc., 2024) Ankit Srivastava; Shruti Mishra; Avadhesh; Anusmita Shekher; Vipin Rai; Anupam Dhasmana; Jayanta Das; Daniele Perenzoni; Renato Iori; Subash C. Gupta
Prohibitin (PHB) is a pleiotropic molecule with a variety of known functions and subcellular locations. PHB's function in breast cancer is poorly understood. Herein, we report that PHB is expressed in cancer types of diverse origin including breast cancer. The cancer patients with changes in PHB were reported to have significantly reduced ‘overall survival’ in comparison to the cases without alterations in PHB. The expression of PHB was increased by H2O2 and also by Moringin (MG), which is an isothiocyanate derived from the seeds of Moringa oleifera. MG interacted with PHB, DRP1, and SLP2 and inhibited the growth of MCF-7 and MDAMB-231 cells. The isothiocyanate triggered apoptosis in breast cancer cells as revealed by AO/PI assay, phosphatidylserine externalization, cell cycle analysis and DAPI staining. MG induced proapoptotic proteins expression such as cytochrome c, p53, and cleaved caspase-7. Further, cell survival proteins such as survivin, Bcl-2, and Bcl-xL were suppressed. A depolarization of membrane potential suggested that the apoptosis was triggered through mitochondria. The isothiocyanate suppressed the cancer cell migration and interacted with NF-κB subunits. MG suppressed p65 nuclear translocation induced by TNF-α. The reactive oxygen species generation was also induced by the isothiocyanate in breast cancer cells. MG also modulated the expression of lncRNAs. Collectively, the functions of PHB in breast cancer growth is evident from this study. The activities of MG against breast cancer might result from its ability to modulate multiple cancer-related targets. © 2023
Neurogenic and angiogenic poly(N-acryloylglycine)-co-(acrylamide)-co-(N-acryloyl-glutamate) hydrogel: preconditioning effect under oxidative stress and use in neuroregeneration
(Royal Society of Chemistry, 2024) Kirti Wasnik; Prem Shankar Gupta; Gurmeet Singh; Somedutta Maity; Sukanya Patra; Divya Pareek; Sandeep Kumar; Vipin Rai; Ravi Prakash; Arbind Acharya; Pralay Maiti; Sudip Mukherjee; Yitzhak Mastai; Pradip Paik
Traumatic injuries, neurodegenerative diseases and oxidative stress serve as the early biomarkers for neuronal damage and impede angiogenesis and subsequently neuronal growth. Considering this, the present work aimed to develop a poly(N-acryloylglycine)-co-(acrylamide)-co-(N-acryloylglutamate) hydrogel [p(NAG-Ac-NAE)] with angiogenesis/neurogenesis properties. As constituents of this polymer modulate their vital role in biological functions, inhibitory neurotransmitter glycine regulates neuronal homeostasis, and glutamatergic signalling regulates angiogenesis. The p(NAG-Ac-NAE) hydrogel is a highly branched, biodegradable and pH-responsive polymer with a very high swelling behavior of 6188%. The mechanical stability (G′, 2.3-2.7 kPa) of this polymeric hydrogel is commendable in the differentiation of mature neurons. This hydrogel is biocompatible (as tested in HUVEC cells) and helps to proliferate PC12 cells (152.7 ± 13.7%), whereas it is cytotoxic towards aggressive cancers such as glioblastoma (LN229 cells) and triple negative breast cancer (TNBC; MDA-MB-231 cells) and helps to maintain the healthy cytoskeleton framework structure of primary cortical neurons by facilitating the elongation of the axonal pathway. Furthermore, FACS results revealed that the synthesized hydrogel potentiates neurogenesis by inducing the cell cycle (G0/G1) and arresting the sub-G1 phase by limiting apoptosis. Additionally, RT-PCR results revealed that this hydrogel induced an increased level of HIF-1α expression, providing preconditioning effects towards neuronal cells under oxidative stress by scavenging ROS and initiating neurogenic and angiogenic signalling. This hydrogel further exhibits more pro-angiogenic activities by increasing the expression of VEGF isoforms compared to previously reported hydrogels. In conclusion, the newly synthesized p(NAG-Ac-NAE) hydrogel can be one of the potential neuroregenerative materials for vasculogenesis-assisted neurogenic applications and paramount for the management of neurodegenerative diseases. © 2024 The Royal Society of Chemistry.
Panchvalkal (polyherbal formulation) mitigates STZ induced type 2 DM by modulating the expression of hexokinase (HX), lactate dehydrogenase (LDH), triphosphate isomerase (TPI)
(Elsevier B.V., 2022) Pratistha Singh; Khushi Verma; Jyoti Dixit; Vipin Rai; Gopeshwar Narayan; Kavindra Nath Tiwari; Anil Kumar Singh; Jasmeet Singh; Kumar Ashutosh
Background: The present study was designed to evaluate the potentiality of Panchvalkal (Ayurvedic formulation) for the treatment of type 2 Diabetes Mellitus. Panchvalkal is polyherbal formulation of bark Ficus benghalensis L., Ficus racemosa L., Ficus religiosa L., Thespesia populnea L. and Ficus lacor Buch-Ham. The formulation was traditionally used as anti-inflammatory, antibacterial and to treat ulcers, wounds, leucorrhoea and other vaginal diseases. Objective: The current study was hypothesized to investigate Panchvalkal aqueous extract protective property against streptozotocin induced Diabetes Mellitus Swiss albino rats and its effects on HX, LDH and TPI genes involved in glycolysis pathway. Method: Aqueous extract of the bark of five medicinal plants were prepared. The aqueous extract of Panchvalkal was analyzed for antioxidant property (TF, TP, DPPH and RP). Diabetes was induced in Swiss albino rats through streptozotocin (60 mgKg-1 bw i.p.) administration. The extract (500 mgKg-1 bw) and glibenclamide (300 mgKg-1 bw) alone and in combination was administrated orally to specific group of animals up to 28 days. Animals were scarified, blood was collected, and liver excised. cDNA was prepared from RNA isolated from the liver. Expression of HX, LDH and TPI were studied through real-time PCR analysis. Result: Panchvalkal aqueous extract was enriched with phenol content (82.84 mg GAE G−1) and flavonoid content (283.52 mg RE G−1) and showed effective antioxidant property. No sign of toxicity and mortality in rats were recorded during toxicity assessment. Rats administrated with Panchvalkal 500 mgKg-1 bw showed significant reduction in blood glucose, total cholesterol, triglycerides, LDL and increase in HDL level. Also the plasma glucose level was significantly (p < 0.05) reduced in comparison to diabetic control. Significant increase in enzymes level such as SGOT and SGPT was observed. Panchvalkal modulated the expression level of Hexokinase (HX), Lactate dehydrogenase (LDH) and Triphosphate isomerase (TPI) genes in STZ induced T2DM. Conclusion: The present study showed that Panchvalkal acts effectively in modulating the mRNA expression of HX, LDH and TPI genes involved in the glycolysis pathway and other metabolic reactions. It significantly increased the glucose uptake and improved the glucose metabolism in T2DM. © 2021 The Authors
Piperlongumine, a piper alkaloid, enhances the efficacy of doxorubicin in breast cancer: involvement of glucose import, ROS, NF-κB and lncRNAs
(Springer, 2022) Nikee Awasthee; Anusmita Shekher; Vipin Rai; Sumit S. Verma; Shruti Mishra; Anupam Dhasmana; Subash C. Gupta
Piperlongumine (PL, piplartine) is an alkaloid derived from the Piper longum L. (long pepper) roots. Originally discovered in 1961, the biological activities of this molecule against some cancer types was reported during the last decade. Whether PL can synergize with doxorubicin and the underlying mechanism in breast cancer remains elusive. Herein, we report the activities of PL in numerous breast cancer cell lines. PL reduced the migration and colony formation by cancer cells. An enhancement in the sub-G1 population, reduction in the mitochondrial membrane potential, chromatin condensation, DNA laddering and suppression in the cell survival proteins was observed by the alkaloid. Further, PL induced ROS generation in breast cancer cells. While TNF-α induced p65 nuclear translocation, PL suppressed the translocation in cancer cells. The expression of lncRNAs such as MEG3, GAS5 and H19 were also modulated by the alkaloid. The molecular docking studies revealed that PL can interact with both p65 and p50 subunits. PL reduced the glucose import and altered the pH of the medium towards the alkaline side. PL also suppressed the expression of glucose and lactate transporter in breast cancer cells. In tumor bearing mouse model, PL was found to synergize with doxorubicin and reduced the size, volume and weight of the tumor. Overall, the effects of doxorubicin in cancer cells are enhanced by PL. The modulation of glucose import, NF-κB activation and lncRNAs expression may have contributory role for the activities of PL in breast cancer. © 2022, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
Poly(N-acryloylglycine-acrylamide) Hydrogel Mimics the Cellular Microenvironment and Promotes Neurite Growth with Protection from Oxidative Stress
(American Chemical Society, 2023) Kirti Wasnik; Prem Shankar Gupta; Sudip Mukherjee; Alagu Oviya; Ravi Prakash; Divya Pareek; Sukanya Patra; Somedutta Maity; Vipin Rai; Monika Singh; Gurmeet Singh; Desh Deepak Yadav; Santanu Das; Pralay Maiti; Pradip Paik
In this work, the glycine-based acryloyl monomer is polymerized to obtain a neurogenic polymeric hydrogel for regenerative applications. The synthesized poly(N-acryloylglycine-acrylamide) [poly(NAG-b-A)] nanohydrogel exhibits high swelling (∼1500%) and is mechanically very stable, biocompatible, and proliferative in nature. The poly(NAG-b-A) nanohydrogel provides a stable 3D extracellular mimetic environment and promotes healthy neurite growth for primary cortical neurons by facilitating cellular adhesion, proliferation, actin filament stabilization, and neuronal differentiation. Furthermore, the protective role of the poly(NAG-b-A) hydrogel for the neurons in oxidative stress conditions is revealed and it is found that it is a clinically relevant material for neuronal regenerative applications, such as for promoting nerve regeneration via GSK3β inhibition. This hydrogel additionally plays an important role in modulating the biological microenvironment, either as an agonist and antagonist or as an antioxidant. Furthermore, it favors the physiological responses and eases the neurite growth efficiency. Additionally, we found out that the conversion of glycine-based acryloyl monomers into their corresponding polymer modulates the mechanical performance, mimics the cellular microenvironment, and accelerates the self-healing capability due to the responsive behavior towards reactive oxygen species (ROS). Thus, the p(NAG-b-A) hydrogel could be a potential candidate to induce neuronal regeneration since it provides a physical cue and significantly boosts neurite outgrowth and also maintains the microtubule integrity in neuronal cells. © 2023 American Chemical Society.
Potential of Long Non-coding RNAs in the Diagnosis and Therapy of Melanoma Skin Cancer
(Springer Nature, 2021) Hitesh Singh Chaouhan; Vipin Rai; Sudarshan Kini; Anusmita Shekher; Anurag Sharma; Subash Chandra Gupta
Skin carcinoma is categorized into melanoma and non-melanoma. Melanoma is among the highly aggressive and deadly forms of skin cancer. Melanoma is frequently associated with metastasis and therapeutic resistance. The combined immunotherapy and targeted therapies have emerged as attractive therapeutic options. However, the efficacy of these therapies is limited to advanced-stage melanoma and those who often acquire resistance. Over the years, the molecular bases of melanoma have been unraveled, which led to establishing specific and reliable biomarkers for the diagnosis, prognosis, and therapy. A good strategy in finding novel cancer targets could include shifting from the protein-translating regions to the genome’s non-coding regions. The non-coding regions constitute approximately 98% of the genome. The microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) are two major classes of non-coding RNAs. Apart from coding RNA’s, lncRNAs have also been attributed to exhibit proto-oncogenic and tumor suppressor roles in various cancers, including melanoma. This chapter summarizes the recent advancement of lncRNAs concerning diagnosis, prognosis, and therapy of melanoma. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2021.