ECT2基因对宫颈癌细胞增殖的影响及其机制

目的 探讨ECT2基因对宫颈癌细胞增殖的影响及其机制。方法 构建ECT2过表达及敲低的宫颈癌细胞株，MTT法检测细胞增殖能力，流式细胞术检测细胞周期。IPA软件查找ECT2的相互作用蛋白，免疫荧光亚细胞定位验证两者间的关系。qPCR及Western blot检测mRNA及蛋白的表达情况。结果 ECT2可能与CDK1相互作用，促进细胞由G₂期进入G₁期，促进细胞增殖。ECT2过表达后宫颈癌细胞中Rac1、Cdc42、CDK1、CyclinB1 mRNA及蛋白的表达水平均升高（均P<0.001）；敲低则作用相反（P<0.05）。结论 ECT2可能通过下游Cdc42/Rac1信号通路，同时与CDK1相互作用促进细胞周期G2向G1转化，进而促进宫颈癌细胞增殖

Effect of ECT2 Gene on Proliferation of Cervical Cancer Cells and Its Mechanism

Objective To study the effect of epithelial cell transformation sequence 2 (ECT2) on the proliferation of cervical cancer cells and its mechanism. Methods We transfected cervical cancer cells HeLa (HeLa-ECT2) with the lentivirus overexpressing ECT2 and the cells SiHa (SiHa-siRNA) and C33a (C33a-siRNA) with the interfering plasmid. MTT assay was performed to detect cell proliferation ability. Flow cytometry was conducted to detect the cell cycle of each group. The IPA database was searched for the interacting proteins of ETC2, and immunofluorescence subcellular localization verified the effect between the two. qPCR and Western blot were carried out to detect the expression of Rac1, Cdc42, CDK1, and Cyclin B1 mRNA and protein in each group of cells. Results ECT2 may interact with CDK1. After ECT2 expression was upregulated, the G₂/M phase of HeLa-ECT2 cells accelerated the transformation to G₁ phase, cell proliferation ability was enhanced, and the expression levels of Rac1, Cdc42, CDK1, and cyclin B1 mRNA and protein all increased (P<0.001); the knockdown of ECT2 expression would reverse the effect (P<0.05). Conclusion ECT2 accelerates G2 phase of cervical cancer cells to G1 phase and promotes cell proliferation by co-localizing with CDK1 through the downstream Cdc42/Rac1 signaling pathway.