Methanolic Neem Bark Extract Restricts Ovarian Cancer Cell Migration And Helps In Re-Establishing The Formation Of Functional Gap Junctional Channels

Vadlamannati, Arunima Vijaya (2021) Methanolic Neem Bark Extract Restricts Ovarian Cancer Cell Migration And Helps In Re-Establishing The Formation Of Functional Gap Junctional Channels. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Arunima Vijaya Vadlamannati (16MS004)) 16MS004.pdf - Submitted Version Restricted to Repository staff only Download (2MB)

Abstract

Neem and its components have widely been studied as anti-cancer agents with clinical usability in multiple cancers. It has also shown anti-carcinogenic activity in many ovarian cancer cell lines having human origin. In our study we focus on Methanolic fraction of Neem Bark Extracts (MNBE) efficiency as an effective anti-carcinogenic agent in mouse ovarian cancer cell lines. The cell line used originated from C57BL/6 mice ovaries that were cultured and spontaneously developed a mutation causing them to become carcinogenic. This cell line was termed ID8. The second cell line used was ID8 cells with VEGF overexpressed in them (ID8-VEGF). Due to this reason, we were capable of developing a syngeneic mice model using C57BL/6 mice. The cell lines were treated with MNBE for 24h for all experiments. The dose for treatment was fixed after performing clonogenic assay. This was decided based on the IC25 value obtained. We studied MNBEs effect on cell migration, gene expression and regulation, Gap Junction Intercellular Communication (GJIC) function and effect on tumorigenicity of ID8-VEGF cells when used in a syngeneic mice model. We performed scratch-assay, real-time PCR, immunofluorescence assay, dye-transfer assay and injecting mice with MNBE-pre-treated tumour cells and untreated cells as a control. This study revealed that MNBE-Treatment is able to inhibit cell migration of moue ovarian cancer cell lines. We have also been able to demonstrate that MNBE is capable of aiding in Cx43 trafficking resulting in an increase in functional gap junctions in both the cell lines. MNBE pre-treated ID8-VEGF cells when injected intraperitoneally decreased its aggressiveness in forming tumour and ascites in C57BL/6 mice as seen by the weight gain. Our study also revealed the in these ovarian cancer cell lines MNBE does not induce the apoptotic pathway to occur. Our study demonstrates certain roles MNBE plays in order to inhibit migration as well as increase cell-cell communication. With this study in place, we conclude that MNBE has effective anti-cancer abilities in these mouse ovarian cancer cell lines.

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