miR-144-3p靶向zeb1调控卵巢癌细胞SKOV3的增殖和凋亡

miR-144-3p targets zeb1 to regulate proliferation and apoptosis of ovarian cancer cell line SKOV3

目的:探讨miR-144-3p是否通过靶向E盒锌指结合蛋白1(zeb1)调控卵巢癌细胞SKOV3的增殖和凋亡。方法:qRT-PCR检测miR-144-3p在卵巢癌细胞和正常人卵巢上皮细胞中的表达差异。在卵巢癌细胞SKOV3中转染miR-144-3p mimics,MTT法测定细胞增殖,PI单染法检测细胞周期,Annexin V-FITC/PI双染法检测细胞凋亡,Western blot检测细胞中cyclinD1、p27、C-caspase-3蛋白表达。生物信息学软件预测miR-144-3p的靶基因可能为zeb1,荧光素酶报告系统鉴定其靶向关系。在卵巢癌细胞SKOV3中共转染miR-144-3p mimics、pcDNA3.1-zeb1,利用上述方法测定细胞增殖、周期和凋亡变化。结果:miR-144-3p在卵巢癌细胞中的表达水平低于正常人卵巢上皮细胞。转染miR-144-3p mimics后的卵巢癌细胞SKOV3增殖能力下降,细胞周期被阻滞在G1期,细胞凋亡增多,细胞中cyclinD1蛋白表达减少,p27、C-caspase-3蛋白表达增加。miR-144-3p靶向调控zeb1表达。pcDNA3.1-zeb1可以逆转miR-144-3p mimics对卵巢癌细胞增殖抑制、周期阻滞和凋亡促进作用。结论:miR-144-3p靶向zeb1抑制卵巢癌细胞SKOV3增殖并诱导细胞凋亡。

Objective:To investigate whether miR-144-3p regulates the proliferation and apoptosis of ovarian cancer cell SKOV3 by targeting E-box zinc finger binding protein 1(zeb1).Methods:qRT-PCR detected the differential expression of miR-144-3p in ovarian cancer cells and normal human ovarian epithelial cells.miR-144-3p mimics was transfected in ovarian cancer cell line SKOV3.MTT assay detected cell proliferation.PI single staining method detected cell cycle.Apoptosis was detected by Annexin V-FITC/PI double staining method.The expression of cyclinD1,p27 and C-caspase-3 protein was detected by Western blot.Bioinformatics software predicted that the target gene for miR-144-3p may be zeb1.The luciferase reporter system identified their targeting relationship.miR-144-3p mimics and pcDNA3.1-zeb1 were transfected in nest cancer cell SKOV3,and cell proliferation,cycle and apoptosis changes were determined using the methods described above.Results:The expression level of miR-144-3p in ovarian cancer cells was lower than that of normal human ovarian epithelial cells.The proliferation of ovarian cancer cell line SKOV3 after transfection with miR-144-3p mimics decreased,the cell cycle was arrested in G1 phase,apoptosis increased,cyclinD1 protein expression decreased,and p27 and C-caspase-3 protein expression increased.miR-144-3p targeted to regulate zeb1 expression.pcDNA3.1-zeb1 can reverse the proliferation inhibition,cell cycle arrest and apoptosis promotion of ovarian cancer cells by miR-144-3p mimics.Conclusion:miR-144-3p targets zeb1 to inhibit proliferation of ovarian cancer cell SKOV3 and induce apoptosis.