TGF-β1促进人子宫颈癌细胞Siha的侵袭、迁移及其机制研究

Investigating the Migration and Invasion of Human Squamous Cervical Carcinoma Line Siha Stimulated by TGF-β1 and Its Underlying Mechanisms

探讨转化生长因子-β1(TGF-β1)对人子宫颈癌细胞Siha侵袭、迁移能力的影响及其可能的分子机制。用5 ng/m L TGF-β1作用Siha细胞72 h,通过倒置显微镜、CCK-8实验、单细胞克隆形成实验、细胞黏附实验和Transwell小室实验分别观察TGF-β1作用前后Siha细胞形态、增殖能力、克隆形成能力、黏附能力、迁移及侵袭能力的改变;Western blot检测TGF-β1作用前后基质金属蛋白酶-2(MMP-2)、MMP-9、VEGF、CFTR、P50、P65、E-cadherin及Vimentin蛋白表达的变化。结果表明,5 ng/m L TGF-β1作用Siha细胞72 h后,Siha细胞出现上皮细胞向间质细胞形态转变、黏附能力减弱、迁移和侵袭能力增强,伴随MMP-2、MMP-9、VEGF、CFTR、P50、P65和Vimentin表达上调,E-cadherin表达下调。该研究表明,TGF-β1可能通过诱导Siha细胞发生上皮–间质转化及上调MMP-2、MMP-9和VEGF的表达,促进Siha细胞的侵袭转移。

This research investigated the influence of transforming growth factor-beta 1 （TGF-β1） on invasion and metastasis of human squamous cervical carcinoma cell line Siha, and might revealed some possible mechanisms. Cultured human cervical carcinoma cell line, Siha, was stimulated with 5 ng/mL of TGF-131 for 72 h. The morpholog- ic changes were observed under inverted microscope. Cell proliferation, clone-formation, adhering ability, migration and invasion ability changes were evaluated by CCK-8, colony-forming, cell adherence and Transwell assays, respec- tively. The expressions of matrix metalloproteinase-2 （MMP-2）, MMP-9, VEGF, CFTR, PS0, P65, E-cadherin and Vimentin were detected by Western blot. We found that after TGF-β1 stimulation, Siha cells converted to an elongat- ed spindle-like shape from a “cobblestone” epithelial structure. The migration and invasion ability of Siha cells were promoted dramatically. The expressions of MMP-2, MMP-9, VEGF, CFTR, P50, P65 and Vimentin were increased and E-cadherin was down-regulated than those of control groups. These results indicated that TGF-β1 might promote the migration and invasion of Siha cells through EMT and up-regulation ofMMP-2, MMP-9 and VEGF.