Ezanee Azlina Mohamad Hanif, Ethar Ahmed Eltayeb Ahmed, Nurul Nadiah Ahmad Daud
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引用次数: 0
Abstract
Background: Triple-negative breast cancer is a challenging type of cancer due to its high recurrence rate, early metastasis, and resistance to therapies. Understanding the key regulatory mechanism to the mentioned oncogenic features is crucial. One such factor being studied is transforming growth factor beta, which has dual roles in promoting tumor growth and controlling cell proliferation. Therefore, this study aims to observe the differential of downstream RhoA/ROCK1 changes and proliferative outcomes in established TNBC cell lines.
Methodology: MDA-MB-231 cells were transduced with lentivirus vectors harboring shRNA targeting different TGFβ isoforms: control cells transduced with non-targeting shRNA (shNT), shTGFβ1, shTGFβ2 and shTGFβ3 respectively. Transduction efficiency and target genes were assessed through RT-qPCR and cell survival capacity was assessed using colony forming assay.
Results: The data showed an inverse TGFβ regulation axis between complete media (CM) and serum-derived (SR) condition. Knockdown of shTGFβ3 was inconclusive, therefore TGFβ3 mode of actions was not further assessed. The interference of baseline growth factor in the complete media condition shown enhanced recovery of cell clonogenic capacity amidst the depletion of TGFβ isoforms. A more stable observation was seen in the SR condition with downregulation of RhoA-ROCK1 expression and reduction of cell survival upon TGFβ isoform depletion. However, compensatory responses were observed in depleted shTGFβ2 whereby induction of clonogenic capacity was displayed in the transduced TNBC cell line.
Conclusion: Serum-derived condition approach provides a consistent readout to measure TGFβ. This provides a better overview of the TGFβ/RhoA/ROCK1 activation axis in TNBC cells. Extended assessments are warranted to fully validate the interaction between this non-canonical network in TNBC, which may be clinically relevant and serves as a new potential therapeutic strategy in TNBC.
期刊介绍：
Molecular Biology Reports publishes original research papers and review articles that demonstrate novel molecular and cellular findings in both eukaryotes (animals, plants, algae, funghi) and prokaryotes (bacteria and archaea).The journal publishes results of both fundamental and translational research as well as new techniques that advance experimental progress in the field and presents original research papers, short communications and (mini-) reviews.
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