下调PKMYT1表达对宫颈癌细胞增殖、迁移与侵袭的影响

Influence of the down-regulated expression of protein kinase membrane associated tyrosine/threonine 1 on the proliferation,migration,and invasion of cervical cancer cell

目的:探究下调膜相关酪氨酸/苏氨酸蛋白激酶(PKMYT1)表达对宫颈癌细胞增殖、迁移和侵袭的影响。方法:利用UALCAN数据库平台分析PKMYT1在宫颈癌组织的表达,实时荧光定量PCR(RT-qPCR)检测PKMYT1在END1/E6E7细胞和宫颈癌细胞的表达。将c-33A细胞、SiHa细胞分为转染对照组(转染siRNA-NC)和siRNA-PKMYT1组(下调PKMYT1表达)。RT-qPCR和Western blot分析无翅型MMTV整合位点家族成员1(WNT1)、β-连环蛋白(β-catenin)和c-Myc的mRNA和蛋白的表达。利用MTT、划痕实验和Transwell实验分析细胞的增殖、迁移和侵袭情况。结果:PKMYT1在宫颈癌组织和细胞表达上调(P<0.05)。siRNA-PKMYT1组c-33A和SiHa细胞中Wnt1、β-catenin和c-Myc表达水平均低于转染对照组,c-33A和SiHa细胞的增殖、迁移和侵袭能力均低于转染对照组(均P<0.05)。结论:沉默PKMYT1可以抑制宫颈癌细胞增殖、迁移和侵袭过程,并降低Wnt/β-catenin信号通路活性,表明PKMYT1在宫颈癌可能发挥抑癌作用。

Objective:To investigate the influence of thedown-regulated expression of protein kinase membrane associated tyrosine/threonine 1(PKMYT1)on the proliferation,migration,and invasion of cervical cancer cell.Methods:UALCAN platform was used to analysis the expression of PKMYT1in the cervical cancer tissues.The quantitative real-time PCR(RT-qPCR)was applied to examine the PKMYT1expression in END1/E6E7cell and in cervical cancer cell.The c-33Acell and SiHa cell were divided into control group(transfection of siRNA-NC)and siRNA-PKMYT1group(downregulation of PKMYT1expression).RT-qPCR and western blot were applied to examine the mRNA and protein expressions of wingless-type MMTV integration site family member 1(WNT1),β-catenin,and c-Myc.MTT,scratch test,and Transwell chamber assays were performed to detect the proliferation,migration,and invasion of the cells.Results:PKMYT1expression in the cervical cancer tissues and in the cells was up-regulated(P<0.05).The expression levels of Wnt1,β-catenin,and c-Myc in the c-33Aand SiHa cells in siRNA-PKMYT1group were significantly lower than those in the control group,and the abilities of proliferation,migration,and invasion of the c-33Aand SiHa cells in siRNA-PKMYT1group were significantly lower than those of the c-33Aand SiHa cells in the control group(all P<0.05).Conclusion:The silence of PKMYT1can inhibit the proliferation,migration,and invasion progressions of the cervical cancer cell,and can decrease the activity of Wnt/β-catenin signaling pathway,which indicates that PKMYT1maybe play a tumor suppressor of the cervical cancer.