The Effection Of Down-regulating ERR? Expression On Endometrial Cancer Cells

Objective To detect the expression of estrogen receptor-related receptor ?(ERR?)interfered by XCT790 and si RNA in RL952, AN3-CA, HEC-1A and HEC-1B cells.Explore the relationship of ERR? expression and proliferation and apoptosis of endometrial cancer cells.Methods The four endometrial cancer cells were cultured in vitro. RL952 and AN3-CA as the type ?endometrial cancer model were ER+,ERR+, while HEC-1A and HEC-1B as the type ?endometrial cancer model were ER-,ERR+. Then, detect the four endometrial cancer cells m RNA and protein expression level of ERR? by quantitative PCR and western blot respectively. Later detect the four endometrial cancer cells m RNA and protein expression level of ERR? interfered by XCT790 using quantitative PCR and western blot.We also detect proliferation and apoptosis of the endometrial cancer cells by MTT and flow cytometry. We successfully constructed ERR? lentivirus si RNA plasmid expression vectors and apply the transient transfection technique in order to suppress the expression of ERR? of endometrial cancer cells. Detect the m RNA and protein expression level of ERR? after si RNA by Real-time fluorescence quantitative PCR and Western blot respectively. Then, the ability of proliferation and apoptosis were compared before and after transfection using MTT and flow cytometry separately.Result1. The relative expression of ERR? m RNA in RL952,AN3-CA cells were remarkbale higher than in HEC-1A and HEC-1B cells.(P <0.05) The protein expression of ERR?in RL952,AN3-CA cells was also much higher than in HEC-1A and HEC-1B cells,that were consistent with the m RNA level.(P <0.05)2. We dealed RL952 and HEC-1A with 10-5M and 10-6MXCT790, respectively( XCT group). Adding normal culture medium group were called negative(NC) and adding DMSO solvent group were called control(CON). The relative expression of ERR?m RNA were inhibited significantly by 10-5MXCT790 in both cells.(P <0.05)3. We treated RL952 and HEC-1A with 10-5MXCT790 for 24 hours and 48 hours,respectively. The relative expression of ERR? m RNA dealed with 10-5MXCT790 were inhibited significantly in both cells for 24 hours and 48 hours.(P <0.05)4. We found the m RNA expression of ERR? in XCT group was significantly down-regulated when treated by 10-5MXCT790 for 24 hours in four endometrial cancer cells, and the inhibition rate were 32.21%RL952, 40.73%AN3-CA,22.76%HEC-A,and 20.32%HEC-1B, respectively.(P<0.05) The protein expression levels of ERR? protein were coincide with m RNA expression level in the experimental groups.5. The growth inhibition ratios of the endometrial cancer cells increased by time when dealed with 10-5MXCT790 for 24 hours, 48 hours, 72 hours, 96 hours(P<0.05). They were RL952(39.67%24h,40.71%48h,43.66%72h,54.25%96h),AN3-CA(9.33%24h,17.71%48h,25.28 %72h,33.12%96h),HEC-1A(20.66%24h,32.65%48h,43.27%72h,44.74%96h),HEC-1B(49.19%24h,57.75%48h,59.43%72h,62.50%96h),respectively.That showed time-dependent.6. The apoptosis rates of the endometrial cancer cells treated with 10-5MXCT were obviously higher than other groups in each cell(P<0.05)7. We constructed the Knock down(KD),Control(CON) and Negative control(NC)group using the ERR? gene lentiviral vector(ERR?-Si RNA-75E), the normal medium and lentiviral vectors NO53 respectively. The m RNA expression of ERR?in KD group was significantly down-regulated in four endometrial cancer cells, and the inhibition rate were 39.77%RL952, 34.01%AN3-CA, 82.82%HEC-1A and 55.78% HEC-1B respectively.(P<0.05) The changes of ERR? protein levels were consistent with the m RNA level.8. The growth inhibition ratios of the endometrial cancer cells after si RNA were remarkably increasing by time when 24 hours, 48 hours, 72 hours, 96 hours(P<0.05).They were RL952(48.14%24h,57.18%48h,85.70%72h,62.22%96h),AN3-CA(28.46%24h,36.21%48h,44.34%72h,30.12%96h),HEC-1A(51.36%24h,52.51%48h,82.01%72h, 49.00%96h), HEC-1B( 38.28%24h, 58.12%48h, 67.46%72h,53.46%96h),respectively. That showed time-dependent.9. The apoptosis rates of KD group in four endometrial cancer cells after si RNA were much higher than CON group and NCgroup in each cell(P<0.05).Conclusion1. XCT790 effected different types endometrial cancers by down-regulating ERR?specifically, which made the proliferation decrease and apoptosis increase in endometrial cancer cells. XCT790, as a specific antagonist, can be used in clinical treatment of endometrial cancer in future.2. ERR? was down-regulated by Si RNA, which made the proliferation decrease and apoptosis increase in endometrial cancer cells.