miR-145调控CCND2蛋白影响卵巢癌细胞的迁移和侵袭

miR-145 Affects the Migration and Invasion of Ovarian Cancer Cells by Regulating CCND2 Protein

目的探讨miK-145对卵巢癌细胞恶性生物学行为的影响及其可能的机制.方法实时定量PCR检测卵巢癌A2780、SKOV3、Hey细胞系中miR-145及细胞周期蛋HD2(cyclin-D2,CCND2)的表达水平,双荧光素酶报告基因检测miR-145及CCND2的调控关系,免疫蛋白印迹实验检测CCND2蛋白的表达含量,流式细胞术检测卵巢癌细胞的增殖周期的变化,CCK-8实验检验SK0V3细胞的增殖能力,Transwell迁移实验检验SKOV3细胞的迁移能力.Transwell侵袭实验检验SK0V3细胞的侵袭能力,结果miR-145在A2780、SKOV3、Hey细胞系中表达含量均下降,并且在SK0V3细胞系中miR-145的表达水平最低;双荧光素酶报告基因结果显乐:miR-145显著抑制SKOV3细胞中CCND2的荧光素酶活性miR-145过表达后,卵巢癌细胞中,G1/G0期细胞数比例增加,而G2/M期细胞比例则减少,CCND2蛋白的表达相应下调,同时削弱了SK0V3细胞增殖、迁移、侵袭能力.结论miR-145町抑制卵巢癌细胞的迁移和侵袭,并靶向下调CCND2的表达。

Objective To investigate the effect of miR-145 on the malignant biological beluivior beluivior of ovarian cancer cells and ils possible mechanism. Methods Real-time quantitative PCR was used to detect the expression levels of niiK-145 and CCND2 in ovarian cancer A2780, SKOV3 and Hey cell lines. The dual luciferase reporter assay was used to detect the regulatory relationship between miR-145 and CCND2 , and the expression of CCND2 protein was detected by Western blotting.The proliferation cycle of ovarian cancer cells was detectef by flow cytometry. The proliferation ability of SKOV3 cells was examined by CCK-8 assay, the migration ability of SKOV3 cells by Transwell migration assay, and I lie inv asion al)ility of SKOV3 cells by Tnmswell invasion assay. RESULTS: The expression levels of miR-145 in A2780, SKOV3 and Hey cell lines were decreased,and the expression level of niiR?145 was the lowest in SKOV3 cell line. Double luciferase reporter assay showed that miR-145 could significantly inhibit the luciferase activity of CCND2 in SKOV3 cells. After overexpression of miR-145 , the proportion of cells in G1/G0 phcise was increased , while the proportion of cells in G2/M phase decreased , and the expression of CCND2 protein was down-regulated,which suggested that the prolifeiation , migration and invasion of SKOV3 cells were weakened. Conclusion miR-145 may inhibit the migmtion and invasion of ovcirian cancer cells by down-regulating the expression of CCND2.