circ-CSPP1靶向miR-433-3p基因调控子宫内膜癌细胞增殖、凋亡以及细胞免疫因子表达的分子机制

circ-CSPP1 targeting miR-433-3p gene regulates the proliferation,apoptosis and cellular immune factors expression of endometrial cancer cells

目的探讨circ-CSPP1靶向miR-433-3p基因调控子宫内膜癌细胞增殖、凋亡以及细胞免疫因子表达的分子机制。方法以子宫内膜癌细胞Ishikawa为研究对象,细胞分为NC组、si-NC组、si-circ-CSPP1组、miR-NC组、miR-433-3p组、anti-miR-NC+si-circ-CSPP1组、anti-miR-433-3p+si-circ-CSPP1组。qRT-PCR检测miR-433-3p和circ-CSPP1相对表达量;MTT法检测细胞增殖;流式细胞术实验检测细胞凋亡;Western blot检测CyclinD1及Cleaved-caspase-3蛋白表达;ELISA法检测TNF-α及IL-6含量;双荧光素酶报告实验检测miR-433-3p和circ-CSPP1靶向关系。结果miR-433-3p在子宫内膜癌和细胞中表达降低,circ-CSPP1相对表达量增加;circ-CSPP1低表达或miR-433-3p过表达可降低CyclinD1蛋白表达、A值,增加Cleaved-caspase-3、凋亡率,减少TNF-α、IL-6含量。circ-CSPP1靶向负调控miR-433-3p的表达,anti-miR-433-3p可以逆转circ-CSPP1低表达对子宫内膜癌细胞增殖、凋亡和细胞免疫因子表达的作用。结论circ-CSPP1低表达可通过增加miR-433-3p抑制子宫内膜癌细胞增殖和促进细胞凋亡,减少细胞免疫因子表达。

This study was performed to investigate the molecular mechanism of circ-CSPP1 targeting miR-433-3p gene in regulating the proliferation,apoptosis and cellular immune factors expression of endometrial cancer cells.Endometrial cancer cell Ishikawa were divided into NC group,si-NC group,si-circ-CSPP1 group,miR-NC group,miR-433-3p group,anti-miR-NC+si-circ-CSPP1 group,anti-miR-433-3p+si-circ-CSPP1 group.qRTPCR was used to detect the relative expression of miR-433-3p and circ-CSPP1;MTT method was used to detect cell proliferation;flow cytometry was used to detect cell apoptosis;Western blot was used to detect the expression of CyclinD1 and Cleaved-caspase-3 proteins;ELISA method was used to detect the levels TNF-αand IL-6;dual luciferase reporter experiment was used to detect the targeting relationship between miR-433-3p and circ-CSPP1.Data showed that the expression of miR-433-3p decreased in endometrial cancer tissues and cells,and the relative expression of circ-CSPP1 increased.Low expression of circ-CSPP1 or overexpression of miR-433-3p could reduce CyclinD1 protein expression and A value,increase Cleaved-caspase-3 expression and apoptosis rate,and reduce the content of TNF-αand IL-6.circ-CSPP1 targeted and negatively regulated the expression of miR-433-3p.Furthermore,anti-miR-433-3p could reverse the effects of low expression of circ-CSPP1 on the proliferation,apoptosis and cellular immune factors expression of endometrial cancer cells.In conclusion,the low expression of circ-CSPP1 can inhibit the proliferation of endometrial cancer cells and promote cell apoptosis by increase miR-433-3p,and reduce the expression of cellular immune factors.