Research On TGF-β1Induced Epithelial To Mesenchymal Transition Of The Human Cervical Cancer Hela Cells

objective: Cervical cancer is one of the three majorgynecological malignant tumors, and cervical cancer is the second commoncancer histological type, which is accounting for about10percent of cervicalcancer. In addition, compared with cervical squamous cell carcinoma, cervicalcancer is easier to divert than cervical squamous cell carcinoma. In recent years,researchers have found that there were important links between epithelialmesenchymal transition (EMT) and tumor development. Transforming growthfactor β1(TGF-β1) has been shown to be an important inducer of epithelialmesenchymal transition. In this study, To investigate the effect of transforminggrowth factor β1(TGF-β1) on Hela cell proliferation, apoptosis and migrationusing epithelial mesenchymal transition (EMT) in vitro. Methods: Humancervical cancer Hela cells cultured in vitro were divided into experimentalgroup and control group. Hela cells were stimulated with differentconcentrations of TGF-β1(0.01,0.1,1,10ng/ml) in experimental group, while,Hela cells cultured in serum-free medium without TGF-β1as control.①Morphological changes of human cervical cancer Hela cells stimulated byTGF-β1(10ng/ml) were observed at different time.②The mRNA expressionsof epithelial mesenchymal transition markers E-cadherin and Vimentin wereevaluated through semi-quantitative RT-PCR assay after Hela cells werestimulated at different concentrations of TGF-β172hour.③The expressions of E-cadherin and Vimentin were determined via immunohistochemistry atdifferent concentrations of TGF-β1.④The influence of proliferation wasestimated by CCK-8method after they were stimulated at differentconcentrations of TGF-β1at72hours.⑤The level of apoptosis wasdetermined via flow cytometry after10ng/ml they were stimulated at differentconcentrations of TGF-β1at72hours.⑥The ability of migration wasdetermined by scratch test after stimulating with10ng/ml TGF-β1. Results:①Morphological changes(extending small prominent part or thansforming cubicshape into spindle polygon) were captured from48h and with time changingsignificantly stimulated with different concentrations of TGF-β1compared withcontrol group. The morphology of most cells in experimental grouptransformed into stromal-like cells, showing elongated fusiform orspindle-shaped, wider cell gap, looser cell connection, and distributed fromcobblestone-like into irregular.②RT-PCR analysis showed that the mRNAexpression of E-cadherin was down regulated, while Vimentin was increasedafter stimulation of TGF-β1, and showed a concentration dependence comparedwith control group(P<0.05).③The cellular immunohistochemistry showedthat the expression of E-cadherin protein gradually decreased, at the same timethe expression of Vimentin protein increased with the increase of concentrationof TGF-β1compared with the control group (P<0.05).④CCK-8tests showedthat different concentrations of TGF-β1has no obvious effect on proliferationof Hela cells for24hours, compared with the control group(P>0.05). But with the stimulation of TGF-β1for48hours or72hours, the proliferation of Helacells was inhibited, and showed a time and concentration dependence,compared with the control group(P<0.05).⑤Flow cytometry showed that therate of apoptosis of human cervical cancer Hela cells gradually increased withthe increase of concentration of TGF-β1for72hours compared with the controlgroup(P<0.05).⑥The ability of migration in Hela cells enhanced afterstimulating with10ng/ml TGF-β1. Conclusion:①TGF-β1might induceepithelial mesenchymal transformation in human cervical cancer Hela cells.②TGF-β1in human cervical cancer Hela cells inhibited proliferation and inducedapoptosis.③TGF-β1might enhance the ability of migration of human cervicalcancer Hela cells.