摘要
目的:探讨RNA结合基序单链相互作用蛋白3反义链3(RBMS3-AS3)对宫颈癌细胞增殖、凋亡和侵袭的影响及作用机制。方法:培养人宫颈永生化细胞Ect1/E6E7和宫颈癌HeLa、Caski和SiHa细胞,实时荧光定量PCR检测细胞中RBMS3-AS3表达水平,Western blot法检测RNA结合基序单链相互作用蛋白3(RBMS3)蛋白水平。将HeLa细胞分为对照组(细胞正常培养)、si-RBMS3-AS3组(转染RBMS3-AS3小干扰RNA)、阴性序列组(转染乱序无意义阴性序列)、si-RBMS3-AS3+si-RBMS3组(共转染RBMS3-AS3和RBMS3的小干扰RNA)和si-RBMS3-AS3+阴性序列组(共转染RBMS3-AS3与乱序无意义阴性序列),四甲基噻唑蓝染色法(MTT)检测细胞增殖率,流式细胞术检测细胞凋亡率,Transwell检测细胞侵袭能力,Western blot检测各组HeLa细胞中细胞周期蛋白D1(Cyclin D1)、B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)和基质金属蛋白酶2(MMP-2)蛋白表达水平。结果:宫颈癌细胞系HeLa、Caski和SiHa中RBMS3-AS3表达水平均高于Ect1/E6E7细胞(P<0.05),而RBMS3蛋白表达低于Ect1/E6E7细胞(P<0.05)。与对照组或阴性序列组相比,si-RBMS3-AS3组HeLa细胞的活性、侵袭细胞数、Cyclin D1、Bcl-2和MMP-2蛋白表达水平降低(P<0.05),细胞凋亡率及RBMS3和Bax蛋白表达水平升高(P<0.05)。与si-RBMS3-AS3+阴性序列组比较,si-RBMS3-AS3+si-RBMS3组HeLa细胞活性、侵袭细胞数及Cyclin D1、Bcl-2和MMP-2蛋白表达水平升高(P<0.05),细胞凋亡率和Bax蛋白表达水平降低(P<0.05)。结论:抑制RBMS3-AS3表达可抑制宫颈癌细胞增殖和侵袭,并促进细胞凋亡,这可能与上调RBMS3表达有关。
OBJECTIVE:To investigate effects from regulation of expression of RBMS3 on proliferation,apoptosis and invasion of cervical cancer cells.METHODS:Immortalized human cervical cells,Ect1/E6E7,and cervical cancer cells:HeLa,Caski and SiHa were cultured,then Real-time quantitative PCR was used to detect expression of RBMS3-AS3 in cells,and Western blot was used to detect protein expression of RBMS3.HeLa cells were divided into control(cells were cultured normally),si-RBMS3-AS3(transfected with RBMS3-AS3 small interfering RNA),negative sequence(transfected with disordered and meaningless negative sequences),si-RBMS3-AS3+si-RBMS3(co-transfected with small interfering RNAs of RBMS3-AS3 and of RBMS3)and si-RBMS3-AS3+negative sequence group(co-transfected with RBMS3-AS3 small interfering RNA and disordered meaningless negative sequences).MTT assay was used to detect proliferation rates of HeLa cells in each group;flow cytometry to detect apoptosis rates;Transwell to detect HeLa cell invasion;Western blot to detect protein expression levels of Cyclin D1,Bcl-2,Bax and MMP-2.RESULTS:Compared with Ect1/E6E7 cells,expression levels of RBMS3-AS3 in cervical cancer cell lines HeLa,Caski and SiHa were significantly increased(P<0.05),and expression levels of RBMS3 protein were decreased(P<0.05).Compared with the negative sequence group,the number of invasive HeLa cells and their expression levels of Cyclin D1,Bcl-2 and MMP-2 proteins in the si-RBMS3-AS3 group were decreased(P<0.05),apoptosis rate and the expression levels of RBMS3 and Bax protein were increased(P<0.05).Compared with the si-RBMS3-AS3+negative sequence group,the number of invasive HeLa cells and their expression levels of Cyclin D1,Bcl-2 and MMP-2 were increased in the si-RBMS3-AS3+si-RBMS3 group(P<0.05),apoptosis rate and Bax protein expression levels were decreased(P<0.05).CONCLUSION:Inhibition of RBMS3-AS3 expression inhibited proliferation and invasion of cervical cancer cells and promoted apoptosis,which might be related to upregulation of RBMS3 expression.
作者
兰莉
陈海丽
徐蕾
LAN Li;CHEN Haili;XU Lei(Obstetrics Department,Shiyan Maternal and Child Health Hospital,Shiyan 442008,Hubei,China)
出处
《癌变．畸变．突变》
CAS
2020年第6期438-443,共6页
Carcinogenesis,Teratogenesis & Mutagenesis
