KLK14基因沉默对OVCAR-3细胞凋亡和迁移能力的影响

Effects of RNA interference targeting kallikrein-related peptidase 14 on apoptosis and migration of OVCAR-3 cells

目的:观察沉默人组织激肽释放酶14(KLK14)基因对人卵巢癌OVCAR-3细胞凋亡和迁移能力的影响。方法:高表达KLK14的OVCAR-3细胞分为3组,实验组以KLK14 shRNA转染细胞,对照组转染对照shRNA,空白组不处理。分别采用RT-PCR和Western blot检测3组细胞KLK14 mRNA和蛋白的表达,采用TUNEL法检测细胞凋亡情况,Transwell小室检测迁移细胞数,Western blot法检测OVCAR-3细胞中Caspase-3、Caspase-9、MMP2和MMP9的表达。结果:与空白组和对照组相比,实验组KLK14 mRNA和蛋白的表达降低(F=187.861和167.374,P均<0.05);OVCAR-3细胞凋亡增加,细胞迁移能力受抑(F=54.313和85.283,P均<0.05);Caspase-3和Caspase-9蛋白的表达增加(F=139.471和91.308,P均<0.05),MMP2和MMP9蛋白的表达减少(F=48.219和61.941,P均<0.05)。结论:下调KLK14的表达可抑制OVCAR-3细胞的恶性行为;KLK14可作为卵巢癌治疗干预的目标。

Aim：To investigate the effect of KLK14 inhibition by RNA interference on the apoptosis and migration in human ovarian carcinoma OVCAR-3 cells.Methods：KLK14 small hairpin RNA（shRNA） was transfected into OVCAR-3 cells with high expression of KLK14（experimental group）.The cells transfected with control shRNA （control group） and the cells without treatment （ blank control group ） were the control .RT-PCR and Western blot were used to detect the expressions of KLK14 mRNA and protein .Apoptosis and migration were observed in OVCAR-3 cells by TUNEL and Transwell chamber assay,and the expressions of Caspase-3,Caspase-9,MMP2 and MMP9 were examined by Western blot .Results：Compared with control group and blank control group ,the expressions of KLK14 mRNA and protein in experimental group were de-creased（F=187.861 and 167.374,P〈0.05）,the apoptosis rate of OVCAR-3 cells was significantly increased ,cell migra-tion was repressed（F=54.313 and 85.283,P〈0.05）,the expressions of Caspase-3 and Caspase-9 protein were increased （F=139.471 and 91.308,P〈0.05）,while those of MMP2 and MMP9 were decreased significantly （F=48.219 and 61.941,P〈0.05）.Conclusion：Inhibition of KLK14 can repress the malignant behavior of OVCAR-3 cells,and KLK14 may serve as a target for ovarian carcinoma therapy .