| Objective:Breast cancer is currently the first malignant tumor that threatens women’s health.It has been reported that the endoplasmic reticulum stress response in breast cancer is activated.Previous studies have found that TMEM16F is highly expressed in breast cancer cells and is related to the proliferation and migration ability of breast cancer cells.This study aims to explore whether TMEM16F promotes the occurrence and development of breast cancer through the endoplasmic reticμlum stress signaling pathway.Methods:The expression levels of GRP78,PERK,p-PERK,ATF4,IRE1,p-IRE1and ATF6 in breast cancer cells MCF-7 and T47D transfected with TMEM16F overexpressing vector and TMEM16F sh RNA vector were detected by Western blot.We use CCK-8 experiment to detect the proliferation of MCF-7 and T47D cells after transfection of the TMEM16F overexpressing vector and the empty vector,and the proliferation of the above-mentioned cells treated with BAPTA-AM and EGTA were detected at the same time.The migration level of breast cancer cells transfected with TMEM16F overexpressing vector and TMEM16F sh RNA vector was verified by cell scratch test.In addition,in MCF-7 and T47D cells transfected with TMEM16F overexpressing vector,the cell proliferation and migration ability of the cells treated with the PERK inhibitor（GSK2606414）was compared through the CCK-8experiment and the cell scratch test.Results:1.In MCF-7 and T47D cells,the expression of GRP78 in the TMEM16F overexpressing group was significantly higher;the expression of GRP78 in the TMEM16F sh RNA group was lower than that in the Scrambled group.2.In MCF-7 and T47D cells,proteins p-PERK/PERK,p-IRE1/IRE1 and ATF4were significantly higher in the TMEM16F overexpressing group;compared with the Scrambled group,p-PERK/PERK,p-IRE1/IRE1 and ATF4 expression was lower in the TMEM16F sh RNA group;and there was no statistically difference in the expression of ATF6 in the above comparison.3.Compared with the control group,the TMEM16F overexpressing group significantly promoted the proliferation of MCF-7 and T47D cells;the TMEM16F overexpression group treated with PERK inhibitor had a weaker effect on the proliferation of cells.4.In MCF-7 and T47D cells,the TMEM16F overexpressing group significantly promoted the migration of MCF-7 cells compared with empty vector group;the TMEM16F sh RNA group had a significantly weaker cell migration level.5.In MCF-7 and T47D cells overexpressing TMEM16F,the migration ability of cells treated with PERK inhibitors were weakened.6.In MCF-7 cells treated with BAPTA-AM and EGTA,the proliferation-promoting effect of TMEM16F on MCF-7 cells was inhibited.Conclusion:1.TMEM16F promotes endoplasmic reticulum stress in breast cancer cells.2.TMEM16F activates PERK pathways and IRE1 protein of endoplasmic reticulum stress,failes to activate the ATF6 protein in breast cancer cells.3.TMEM16F promotes the proliferation and migration of breast cancer cells MCF-7 and T47D.4.PERK protein in endoplasmic reticulum stress mediates TMEM16F to promote the of breast cancer cells MCF-7 and T47D.5.The proliferation effect of TMEM16F on breast cancer cells depends on Ca2+. |
