Banita Verma

All Publications

• Presence of key cholinergic enzymes in human spermatozoa and seminal fluid†. Biology of reproduction Thakur, B., Hasooni, L. P., Gera, R., Mitra, S., Björndahl, L., Darreh-Shori, T. 2024; 110 (1): 63-77

  Abstract

  Little is known about the non-neuronal spermic cholinergic system, which may regulate sperm motility and the acrosome reaction initiation process. We investigated the presence of the key acetylcholine (ACh)-biosynthesizing enzyme, choline acetyltransferase (ChAT), and the acetylcholine-degrading enzymes, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and two ACh-receptors in human spermatozoa and seminal plasma. Fresh ejaculates were used for intra- and extracellular flow cytometric analysis of ChAT, AChE, BChE, and alpha-7-nicotinic and M1-muscarinic ACh-receptors in sperm. For determining the source of soluble enzymes, frozen seminal samples (n = 74) were selected on two bases: (1) from vasectomized (n = 37) and non-vasectomized (n = 37) subjects and (2) based on levels of alpha-glucosidase, fructose, or zinc to define sample subgroups with high or low fluid contribution from the epididymis and seminal vesicle, and prostate, respectively. Flow cytometric analyses revealed that ChAT was expressed intracellularly in essentially all spermatozoa. ChAT was also present in a readily membrane-detachable form at the extracellular membrane of at least 18% of the spermatozoa. These were also highly positive for intra- and extracellular BChE (>83%) and M1 (>84%) and α7 (>59%) ACh-receptors. Intriguingly, the sperm was negative for AChE. Analyses of seminal plasma revealed that spermatozoa and epididymides were major sources of soluble ChAT and BChE, whereas soluble AChE most likely originated from epididymides and seminal vesicles. Prostate had relatively minor contribution to the pool of the soluble enzymes in the seminal fluid. In conclusion, human spermatozoa exhibited a cholinergic phenotype and were one of the major sources of soluble ChAT and BChE in ejaculate. We also provide the first evidence for ChAT as an extracellularly membrane-anchored protein.

  View details for DOI 10.1093/biolre/ioad127

  View details for PubMedID 37741056

  View details for PubMedCentralID PMC10790344

• Effect of aspirin on the TNF-α-mediated cell survival and death pathways in breast cancer. Journal of basic and clinical physiology and pharmacology Thakur, B., Saha, L., Dahiya, D., Bhatia, A. 2023; 34 (1): 91-102

  Abstract

  Aspirin is an anti-inflammatory drug commonly used as an analgesic and in cardiovascular disorders. However, many studies have highlighted its anti-cancer properties, especially in colorectal, lung, head and neck, and breast cancers. In this work, we tried to study the effect of aspirin on the TNF-α-mediated cell survival and death pathways in two cell lines representing two different subtypes of breast cancer. TNF-α-mediated stimulation of a cell can result in its proliferation via the NF-κB pathway or its death via either apoptosis or a programmed form of necrosis called necroptosis. The latter is believed to come into the picture only when apoptosis is inhibited.In this work, we studied the effect of aspirin on the TNF-α-mediated cell survival pathway and observed a decrease in expression of the NF-κB pathway regulators, its nuclear translocation, and phosphorylation in a dose-dependent manner. The effect of aspirin on the TNF-α-mediated cell death showed significant cytotoxicity at the higher doses (5-20 mM) of aspirin in both the breast cancer cell lines. The effect of aspirin on necroptosis was investigated after stimulating the cells with TNF-α and inhibiting apoptosis using Z-VAD-FMK.Though no significant effect was noted in breast cancer cell lines, the above protocol successfully induced necroptosis in L929, i.e., a positive control cell line for necroptosis having an intact necroptosis machinery. Even when combined with the chemotherapeutic drugs, the above regime failed to induce any significant necroptosis in breast cancer cells but was found effective in L929.Overall, the findings show that while aspirin has the potential to inhibit the TNF-α-mediated cell survival pathway, it does not help sensitize breast cancer cells to necroptotic cell death induction.

  View details for DOI 10.1515/jbcpp-2022-0112

  View details for PubMedID 36378010

• Synthesis, single crystal X-ray, DFT, spectroscopic, molecular docking studies and<i> in</i><i> vitro</i> biological evaluation of compound N-benzyl-4-(4-chlorophenyl)-2-oxobutanamide JOURNAL OF MOLECULAR STRUCTURE Das, B., Baidya, A. K., Devi, B., Rom, T., Paul, A., Thakur, B., Darreh-Shori, T., Kumar, R. 2023; 1276

  View details for DOI 10.1016/j.molstruc.2022.134782

  View details for Web of Science ID 000912942100001

• Relative refractoriness of breast cancer cells to tumour necrosis factor-α induced necroptosis. Clinical and experimental pharmacology & physiology Thakur, B., Saha, L., Bhatia, A. 2022; 49 (12): 1294-1306

  Abstract

  Necroptosis, a recently identified programmed cell death pathway, has attracted attention as an alternative route to target apoptosis-resistant cancer cells. The status of the necroptosis pathway in different subtypes of breast cancer has not been well explored. Stimulating the cells by TNF-α can trigger cell survival or death depending on the combination of downstream players involved. In this work, we attempted to induce necroptosis in them using a combination of TNF-α and Z-VAD-FMK with and without chemotherapy. Cell viability, apoptosis, and necroptosis were assessed using MTT and Annexin-V/PI assays, respectively. Gene and protein expression was analysed by qPCR and immunophenotyping. Both the cell lines were resistant to induction of cell death by necroptosis. There was no enhancement in cell death when chemotherapeutic drugs were combined with necroptosis induction. Expression studies showed reduced translational expression of key necroptosis molecules like RIP kinases and MLKL in breast cancer cells compared to positive control cell line L929. Also, cell survival molecules were expressed more in MDA-MB-231 in contrast to death pathway molecules which were expressed more in T47D cells. In this work, the two breast cancer cell lines were observed to be resistant to TNF-α induced necroptosis with or without chemotherapy. Expression of key necroptosis players revealed relative insufficiency of the molecular machinery involved in the above pathway. In our opinion this may be the cause for resistance to necroptosis and novel strategies to upregulate these molecules need to be developed to sensitize the breast cancer cells towards cell death by necroptosis.

  View details for DOI 10.1111/1440-1681.13711

  View details for PubMedID 36054417

• Effect of dietary AGEs on the transcriptional profile of peripheral blood lymphocytes APPLIED FOOD RESEARCH Sudha, M., Banita, Ram, A. K., Bhatia, A. 2022; 2 (1)

  View details for DOI 10.1016/j.afres.2022.100086

  View details for Web of Science ID 001136207800006

• Do Different Stemness Markers Identify Different Pools of Cancer Stem Cells in Malignancies: A Study on ER+ and ER-Breast Cancer Cell Lines. Pathology oncology research : POR Chopra, S., Goel, S., Thakur, B., Bhatia, A. 2020; 26 (1): 371-378

  Abstract

  In view of popularity of cancer stem cell (CSC) model all events in evolution of cancer are being explained in that context. Breast cancer is first solid tumor in which CSCs were identified. We aimed to compare stemness profile of two major subtypes [Estrogen receptor positive (ER+) and negative (ER-)] breast cancer using different sets of markers. Expression of CD44/CD24, CK/Vimentin, E-Cadherin/Fibronectin and percentage of side population (SP) was studied in ER+ (T47D) and ER- (MDA-MB-231) cell lines by flow cytometry. Breast CSCs (BCSCs) were sorted using CD44+/CD24-/low expression and SP analysis and cultured. BCSCs were then compared with Non-CSCs (NCSCs) for response to drugs (Paclitaxel and Cisplatin), Ki67 and ER expression. Results showed higher expression of stemness markers (CD44+/CD24-/low, CK+/Vimentin+ and E-Cadherin-/FibrinectinF+) in MDA-MB-231 cells. Percentage SP representing BCSCs was found to be significantly more in later (3.20 ± 0.002 cf. T47D 1.25% ± 0.0007). BCSCs were found to be more resistant to drugs as compared to NCSCs in both cell lines. ER expression was weak in BCSCs sorted from T47D as compared to NCSCs. Ki67 was expressed in both BCSCs and NCSCs. Differences in expression of stemness markers help to explain aggressive behavior, higher recurrence rate and metastatic potential of MDA-MB-231 cells. However, no correlation amongst different markers used suggests that they may be identifying varied populations of cells in tumor hierarchy. A weak ER expression in BCSCs may be strategy used by BCSCs to escape effect of hormone therapy in ER+ breast cancers.

  View details for DOI 10.1007/s12253-018-0503-8

  View details for PubMedID 30361903

• Programmed necrosis and its role in management of breast cancer. Pathology, research and practice Thakur, B., Kumar, Y., Bhatia, A. 2019; 215 (11): 152652

  Abstract

  Breast cancer is one of the major causes of cancer related deaths in women worldwide. A major factor responsible for treatment failure in breast cancer is the development of resistance to commonly used chemotherapeutic drugs leading to disease relapse. Several studies have shown dysregulation of molecular machinery of apoptosis, the major programmed cell death pathway in breast malignancies. Thus, there is an unmet need to search for an alternative cell death pathway which can work when apoptosis is compromised. Necroptosis or programmed necrosis is a relatively recently described entity which has attracted attention in this context. Classically, even in physiological conditions necroptosis is found to act if apoptosis is not functional due to some reason. Recently, more and more studies are being conducted in different malignancies to explore the possibility and utility of inducing cell death by necroptosis. The present review describes the key molecular players involved in necroptotic pathway and their status in breast cancer. In addition, the research done to utilize this pathway for treatment of breast cancer has also been highlighted.

  View details for DOI 10.1016/j.prp.2019.152652

  View details for PubMedID 31570277