跨膜蛋白48对宫颈癌细胞增殖、侵袭、迁移及Yes相关蛋白通路影响研究

The effect of transmembrane protein 48 on the proliferation,invasion,migration and Yes-associated protein pathway of cervical cancer cells

目的 探讨跨膜蛋白48(TMEM 48)对宫颈癌细胞增殖、侵袭、迁移及Yes相关蛋白(YAP)通路的影响。方法 用实时荧光定量聚合酶链式反应(PCR)法和蛋白免疫印迹法分别检测35例宫颈癌组织及其配对的癌旁组织、正常宫颈上皮细胞(H8)和宫颈癌细胞系(SiHa、HeLa、CaSki、C33A)的TMEM 48基因及蛋白表达水平。分别用TMEM 48 si-RNA(沉默组)或者si-control(对照组)转染HeLa和CaSki细胞。用CCK-8法检测细胞增殖;Transwell小室实验检测细胞侵袭和迁移。实时荧光定量PCR法检测TMEM 48基因表达水平;蛋白免疫印迹法检测细胞TMEM 48、YAP、pYAP蛋白表达水平,检测细胞核和细胞质中YAP蛋白表达水平。结果 与癌旁组织比较,癌组织中TMEM 48基因和蛋白表达水平均较高,差异有统计学意义(P<0.05)。与正常宫颈上皮细胞H8比较,宫颈癌SiHa、HeLa、CaSki、C33A细胞系的TMEM 48基因和蛋白表达水平较高,差异有统计学意义(P<0.05)。在HeLa和CaSki细胞中,对照组CCK-8实验72 h时的细胞增殖活性、侵袭细胞数、迁移细胞数均低于沉默组,差异有统计学意义(P<0.05)。沉默组TMEM 48基因和蛋白相对表达量均明显低于对照组,差异有统计学意义(P<0.05)。与对照组比较,沉默组pYAP蛋白表达水平、pYAP/总YAP、YAP蛋白的核质比均较低,差异有统计学意义(P<0.05)。结论 宫颈癌细胞和组织中TMEM 48表达水平较高,沉默TMEM 48表达可以抑制宫颈癌细胞的增殖、侵袭、迁移及YAP通路激活。

Objective To explore the effect of transmembrane protein 48(TMEM 48)on the proliferation,invasion,migration and Yes-related protein(YAP)pathway of cervical cancer cells.Methods TMEM 48 mRNA and protein in 35 cases of cervical cancer tissues and their paired adjacent tissues,normal cervical epithelial cells(H8)and cervical cancer cell lines(SiHa,HeLa,CaSki,C33A)detected by real-time fluorescent quantitative polymerase chain reaction(PCR)and Western-blot.HeLa and CaSki cells were transfected with TMEM 48 si-RNA(silence group)or si-control(control group)respectively.The CCK-8 method was used to detect cell proliferation;the Transwell chamber experiment was used to detect cell invasion and migration.Real-time fluorescent quantitative PCR method was used to detect TMEM 48 gene expression level;Western-blot was used to detect cell TMEM 48,YAP,pYAP protein expression levels,and cell nucleus and cytoplasmic YAP protein expression levels.Results TMEM 48 gene and protein expression levels in cancer tissues were higher than those of para-carcinoma tissue,the difference was statistically significant(P<0.05).TMEM 48 gene and protein expression levels in cervical cancer cell lines(SiHa,HeLa,CaSki,C33A)were higher than those of normal cervical epithelial cells(H8),the difference was statistically significant(P<0.05).In HeLa and CaSki cells,the cell proliferation activity,the number of invaded cells,and the number of migrating cells in the CCK-8 experiment of the control group at 72 h were lower than those of the silent group,the difference was statistically significant(P<0.05).The relative expression of TMEM 48 gene and protein in the silence group was significantly lower than those of the control group,the difference was statistically significant(P<0.05).The relative expression of TMEM 48 gene and protein in the silence group was significantly lower than those of the control group,the difference was statistically significant(P<0.05).The expression level of pYAP protein,pYAP/total YAP,and the ratio of nucleus to cytoplasm of YAP protein in the silent group were lower than those of the control group,the difference was statistically significant(P<0.05).Conclusion The expression level of TMEM 48 in cervical cancer cells and tissues is high.Silencing the expression of TMEM 48 can inhibit the proliferation,invasion,migration and YAP pathway activation of cervical cancer cells.