The Effect Of TGF-β1 On The Biological Behaviors Of Human Cervical Cell Line SiHa

Objective:Cervical cancer is the most common malignant tumor in gynecology. Metastasis is an important factor that influences the patients’prognosis. In this study, cultured human cervical carcinoma cell line, SiHa, was stimulated with 5ng/mL of Transforming growth factor-beta 1(TGF-β1) for 72 h. The morphologic 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, respectively. Analyse the correlation between the migration and invasion ability and epithelial-mesenchymal transition (EMT) of SiHa cells; clarify the possible mechanism that TGF-β1 induced EMT of SiHa cells. This study will thus provide theoretical basis for therapy of cervical cancer.Methods:1:Studying the influence of TGF-β1 on the biological behaviors of SiHa cells. After 5ng/mL TGF-β1 treated SiHa cells for 72 h, the morphologic 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, respectively.2:Studying the influence on the degradation of extracellular matrix (ECM) and angiogenesis, after TGF-01 treated SiHa cells for 72 h. The expressions of Matrix metalloproteinase-2(MMP-2), MMP-9 and Vascular endothelial growth factor(VEGF) after TGF-betal treated were detected by Western blots.3:Analyze the correlation between the enhancement of migration and invasion ability and epithelial-mesenchymal transition (EMT) of SiHa cells; Clarify the possible mechanism how EMT induced by TGF-β1. The expressions of cystic fibrosis transmembrane conductance regulator (CFTR), P50, P65, E-cadherin and Vimentin after TGF-betal treated were detected by Western blots.Results:1:After TGF-β1 stimulation, SiHa cells were converted from a "cobblestone" epithelial structure into an elongated spindle-like shape, the migration and invasion ability of Siha cells were promoted dramatically, but no significant effect on SiHa cells proliferation and clone-formation.2:After TGF-β1 treated SiHa cells for 72 h, the expressions of MMP-2, MMP-9 and VEGF were increased than those of control groups.3:After TGF-β1 treated SiHa cells for 72 h, the expressions of CFTR, P50, P65 and vimentin were increased and E-cadherin was down-regulated than control groups.Conclusions:1:This study showed that TGF-β1 is related to cervical cancer progress and distant metastasis in vitro. TGF-β1 is mainly involved in SiHa cells adhesion, invasion and metastasis.2:TGF-β1 might promote the migration and invasion of SiHa cells through EMT and up-regulation of MMP-2, MMP-9 and VEGF.