Effect Of IL-17on Proliferation Of Human Cervical Carcinoma Cell Line Hela In Vitro And Its Mechanism

Objective: Discussing the effect of interleukin17(IL-17) on the occurrenceand development of cervical carcinoma and finding out the mechanism for therealization of such function to provide more experimental basis for its clinicaltreatment.Methods: First, human cervical carcinoma cell line HeLa was cultured inDMEM medium(high glucose) with IL-17at different concentrations (0ng/ml、1ng/ml、10ng/ml、50ng/ml、100ng/ml) in vitro, then finish the detections below:(1)After24h of cell culture, MIT assay was performed to evaluate cell proliferation;(2) After stimulatation of the induced apoptosis cells with il-17by24h, we detectedthe cell apoptosis by flow cytometry.;(3) After48h of cell culture, we collected thecells and extracted the total RNA and then semi-quantitative RT-PCR was used todetect the VEGF mRNA;(4) After48h of cell culture, we collected the cells, andextracted the intracellular protein, whereafter we used Western blot assay to detectVEGF protein.Results:(1) Treated the cell line HeLa with IL-17at1ng/ml、10ng/mland100ng/ml for24h, no significant difference was observed between IL-17-treatedand control groups while the proliferation was obviously promoted in group with50ng/ml IL-17and statistical difference was detected.(2) The apoptotic rate of cellsin the late stage clearly dropped for50ng/ml and100ng/ml and presented statisticaldifference (P<0.01) after24h co-culture with IL-17while compared with the controlgroup,and the total cell apoptosis rate of both50and100ng/ml IL-17treatmentgroup had significantly decreased compared with the normal control group (P<0.05,P<0.01respectively),there was an increase of the apoptotic rate in the early stage for50ng/ml while with no statistical difference and yet a decrease for100ng/ml with no statistical difference too.(3) After48h co-culture with IL-17, cells expressed VEGFmRNA for both50ng/ml and100ng/ml were higher than control group, especiallyfor50ng/ml (P<0.05).(4) For treated group, there was a higher level of VEGFprotein expression than that in control group after48h co-culture with IL-17especially for50mg/ml (P<0.05).Conclusions:(1) Exogenous IL-17at concentration50ng/ml can promote theproliferation of human cervical carcinoma cell line HeLa in vitro;(2) IL-17with50ng/ml and100ng/ml could inhibit the apoptosis of human cervical carcinoma cellline HeLa (the apoptotic rate in both late stage and the total distinctly decreased incontrast with the control group) and the inhibitory effect enhanced with higherconcentration IL-17, presenting a dose-dependent manner;(3)50ng/ml IL-17couldincrease the expression of VEGF mRNA of human cervical carcinoma cell lineHeLa;(4)50ng/ml IL-17could increase the expression of VEGF protein of humancervical carcinoma cell line HeLa;(5) The promotion effect of IL-17on the processof neoplasia might be through inhibiting the apoptosis of tumor cells and increasingthe expression of VEGF which promoted blood vessel formation in both mRNA andprotein.