GBX2过表达促进人宫颈癌SiHa细胞增殖、迁移及侵袭

摘要] 目的：探讨GBX2 基因在人宫颈癌SiHa 细胞增殖和侵袭转移中的作用及其机制。方法：应用质粒转染技术，分别将过表达GBX2 基因重组质粒pCMV6-entry-GBX2（实验组）及空载体质粒pCMV6-entry（阴性对照组）转染到宫颈癌SiHa 细胞中，用WST-1 法、集落形成实验、流式细胞术分别检测转染细胞的增殖、集落形成和细胞周期，用划痕愈合实验、Transwell 实验检测细胞的迁移、侵袭能力，用ELISA 法检测细胞培养上清中IL-6 的表达水平，用WB检测EMT相关蛋白的表达变化并探讨其可能的作用机制。结果：与SiHa/pCMV6 组相比，上调GBX2 表达后：（1）SiHa/GBX2 组细胞的增殖、集落形成、迁移和侵袭能力明显增强（均P<0.01），G0/G1 期的细胞比例减少、S期与G2/M期的细胞比例增加（均P<0.01）；（2）SiHa/GBX2 组细胞EMT相关蛋白上皮钙黏蛋白表达水平下降，神经钙黏蛋白、波形蛋白和snail 表达水平上调（均P<0.01）；（3）SiHa/GBX2 组细胞培养上清中IL-6 的表达水平明显增高（P<0.01）；（4）SiHa/GBX2 组细胞STAT3 磷酸化水平增强，并能被STAT3 抑制剂S31-201 抑制（P<0.01）。结论：GBX2可能通过IL-6/STAT3 通路诱导宫颈癌SiHa 细胞EMT，从而促进宫颈癌细胞的增殖、迁移和侵袭能力。

GBX2 over-expression promotes proliferation, migration and invasion of human cervical carcinoma SiHa cells

Abstract] Objective: To investigate the effects of GBX2 gene on the proliferation, migration and invasion of human cervical carcinoma SiHa cells and to explore the mechanism. Methods: Recombinant plasmid over-expressing GBX2 gene (pCMV6-entry-GBX2,experimental group) and empty vector plasmid (pCMV6-entry, negative control group) were transfected into cervical cancer SiHa cells by plasmid transfection technique. The proliferation, colony formation and cell cycle of transfected cells were detected by WST-1 method,Colony formation assay and flow cytometry, respectively. The cell migration and invasion were detected by wound healing assay and Transwell assay. The expression level of IL-6 in cell culture supernatant was detected by ELISA. WB was used to detect the expression changes of EMT-related proteins and to explore its possible mechanism. Results: Compared with the SiHa/pCMV6 negative control group, after up-regulation of GBX2, (1) the proliferation, colony formation, migration and invasion of SiHa/GBX2 cells in the experimental group were significantly enhanced (all P<0.01); The proportion of cells in G0/G1 phase decreased while the proportion of cells in S phase and G2/M phase increased (all P<0.01); (2) the expression of E-cadherin decreased, and the expressions of N-cadherin, vimentin and snail increased (all P<0.01); (3) the expression of IL-6 in the culture supernatant of SiHa/GBX2 cells was significantly up-regulated (P<0.01); (4) STAT3 phosphorylation in SiHa/GBX2 cells was enhanced, and could be inhibited by STAT3 inhibitor S31-201 (P<0.01). Conclusion: GBX2 may induce EMT of cervical cancer SiHa cells through IL-6/STAT3 pathway, thereby promoting the proliferation, migration and invasion of cervical cancer cells.