Mechanism Of TRPM7 On The Invasion And Metastasis Of Ovarian Cancer Cells Regulation Of Epithelial-Mesenchymal Transition

Objective: We have previously found that TRPM7(Transient receptor potential melastatin 7)can promote the invasion and migration of ovarian cancer cells,but the mechanism has not yet been elucidated.The aim of this study is to investigate the role and molecular mechanism of TRPM7 in the regulation of ovarian cancer cell line EMT via[Ca2+]i-PI3K/AKT signaling axis,and to provide new molecular targets and new strategies for anti-metastasis treatment of ovarian cancer.Method: In this study,epithelial ovarian cancer cell lines with stable silencing of TRPM7 were established respectively in SKOV3 and OVCAR3.The method of scratching,Transwell invasion and migration,tail vein injection of lung metastasis in nude mice and other methods was used to investigate TRPM7 in epithelial ovary in vitro and in vivo.The effects of cancer cell invasion and migration were observed by morphological observation,flow cytometry,Western Blot,qRT-PCR,etc.,or by inhibiting intracellular calcium signaling,or by administering PI3K/AKT activator/inhibition.Effects of treatments on EMT-related markers and calcium signaling in epithelial ovarian cancer cells,and the expression of PI3K/AKT signaling pathway molecules.Results: 1.This study successfully constructed a stable and silent epithelial ovarian cancer cell line of TRPM7.After in vivo and in vitroshowed silencing of TRPM7 expression,the invasion and metastasis ability of epithelial ovarian cancer cells was significantly weakened and EMT changes were significantly inhibited.And then using Fluo-8-AM labeled calcium ions,laser confocal and flow cytometry techniques,silencing TRPM7 Expression can significantly attenuate intracellular calcium signaling.2.In this study,laser confocal and flow cytometry were used to find that the calcium chelating agent BAPTA-AM can effectively block the calcium signal in SKOV3 and OVCAR3 cells and inhibit the invasion and metastasis of epithelial ovarian cancer cells.When SKOV3 and OVCAR3 cells were treated with BAPTA-AM,they activated or inhibited the PI3K/AKT signaling pathway,respectively.The results of invasion and migration assay showed that the inhibitory(activated)PI3K/AKT signaling pathway had enhanced(cancelled)effects,suggesting that calcium signaling can be Invasion and metastasis of epithelial ovarian cancer cells are affected by the PI3K/AKT pathway.3.This study blocked the calcium signal in TRPM7-stabilized epithelial ovarian cancer cells by BAPTA-AM.And enhanced the inhibitory effect of TRPM7 silencing on the invasion and migration of epithelial ovarian cancer cells.4.In vivo and in vitro experiments showed that silencing TRPM7 expression can inhibit PI3K-mediated Akt phosphorylation by [Ca2+]i,thereby inhibiting EMT of epithelial ovarian cancer cells.Conclusion: 1.[Ca2+]i promotes EMT,invasion and metastasis of epithelial ovarian cancer cells by activating the PI3K/AKT pathway.2.Silencing TRPM7 can inhibit the invasion and metastasis of epithelial ovarian cancer cells by inhibiting EMT mediated by[Ca2+]i-PI3K/AKT signaling pathway.