干细胞转录因子Oct4及Sox2对宫颈癌细胞成瘤、迁移和浸润能力的影响

Effects of Stem Cell Transcription Factors Oct4 and Sox2 on the Occurrence,Migration and Invasion of Cervical Cancer Cells

目的探讨Oct4和Sox2对宫颈癌细胞(以下简称HeLa细胞)成瘤、迁移和浸润能力的影响。方法复苏并培养HeLa细胞,分别构建Oct4及Sox2过表达质粒及空载质粒,采用脂质体方法转染至HeLa细胞,按转染质粒的不同分为4组(正常对照组、Oct4过表达组、Sox2过表达组和过表达空载组),铺至6孔板中,每组一个孔(细胞量为1×10^(6)),转染后48h进行PCR验证,转染成功后采取细胞划痕实验检测细胞迁移能力、Transwell检测细胞侵袭能力及成球实验检测细胞成瘤能力,并采用统计学方法对各组间差异进行分析。结果PCR验证结果显示,Oct4及Sox2过表达组HeLa细胞中Oct4(5376±208.7,2.429±0.1816)和Sox2(133±23.1,1991842±210370)表达显著高于空载组(2.182±0.5183,69.27±11.94)及对照组(0.8582±0.1579,0.7932±0.2134),差异均有统计学意义(F=662.8,89.64,均P<0.0001),表明转染成功;细胞划痕实验结果显示,HeLa细胞中Oct4过表达组迁移率(25.00±5.56)及Sox2(27.00±1.73)低于空载组(36.67±0.57)及对照组(35.67±0.57),差异有统计学意义(F=12.21,P<0.05);细胞侵袭实验结果显示,HeLa细胞中Oct4过表达组侵袭数量(378.3±97.59)及Sox2(374.3±43.89)表达显著低于空载组(552.7±41.48)及对照组(515.7±57.4),差异有统计学意义(F=6.22,P<0.05);成球实验结果显示,HeLa细胞中Oct4过表达组成球数量(8.667±2.082)及Sox2(8.333±1.155)低于空载组(14.67±2.517)及对照组(13.67±1.528),差异有统计学意义(F=9.11,P<0.05)。结论Oct4和Sox 2作为肿瘤干细胞的转录因子具有抑制宫颈癌细胞的侵袭、转移及生长的作用,为宫颈癌的早期治疗和预防提供新方向。

Objective To investigate the effects of Oct4 and Sox2 on tumorigenesis,migration and infiltration of human cervical cancer cells(hereinafter referred to HeLa cells).Methods The HeLa cells were resuscitated and cultured,and the Oct4 and Sox2 overexpression plasmids and empty plasmids were constructed respectively.The transfected cells were transfected into HeLa cells by liposome method and divided into four groups(normal control group,Oct4 overexpression group,Sox2 overexpression group,overexpression no-load group)according to the different transfected plasmids and spread into six-well plates,where the cell volume per group was 1×10^(6).PCR verification was performed 48h after transfection.After successful transfection,cell migration ability was detected by cell scratch test,and cell invasion ability and tumorigenesis ability were detected by Transwell test.The differences between groups were analyzed using statistical methods.Results PCR verification results showed that the overexpression of Oct4 and Sox2 in HeLa cells(5376±208.7,2.429±0.1816)and Sox2(133±23.1,1991842±210370)were significantly higher than those in the no-load group(2.182±0.5183,69.27±11.94)and the control group(0.8582±0.1579,0.7932±0.2134),the differences were statistically significant(F=662.8,89.64,P<0.0001),indicating that the transfection was successful.The HeLa scratch test results showed that the mobility(25.00±5.56)and Sox2 Sox2(27.00±1.73)of the Oct4 overexpression group were lower than those in the no-load group(36.67±0.57)and the control group(35.67±0.57),the difference was statistically significant(F=12.21,P<0.05).The results of HeLa invasion experiments showed that the number of invasions(378.3±97.59)and Sox2(374.3±43.89)expressions in the Oct4 overexpression group were significantly lower than those in the no-load group(552.7±41.48)and the control group(515.7±57.4),the difference was statistically significant(F=6.22,P<0.05).The number of Oct4 overexpressing constituent spheres(8.667±2.082)and Sox2(8.333±1.155)in HeLa cells in the pelletization experiment were lower than those in the no-load group(14.67±2.517)and the control group(13.67±1.528),the difference was statistically significant(F=9.11,P<0.05).Conclusion As transcription factors of tumor stem cells,Oct4 and Sox-2 can inhibit the invasion,metastasis and growth of cervical cancer cells,thus providing a new direction for the early treatment and prevention of cervical cancer.