Effect Of Rapamycin On Human Cervical Cancer Hela Cells And HIF-1α, VEGF Expression

Objective: In this study, different concentrations of rapamycin to deal with in vitro cultured human cervical cancer Hela cells, observation of rapamycin on the proliferation of Hela cells, as well as HIF-1αand VEGF expression. Drug treatment for cervical cancer to provide new ideas and methods for further study of rapamycin in the treatment of cervical cancer could be applied to provide an important theoretical basis.Methods: In vitro cultured human cervical cancer Hela cell line sub-experimental group and control group, experimental group, as the role of different concentrations of rapamycin for 72 hours. MTT assay of cell proliferation inhibition rate, describe the inhibition rate and the relationship between drug concentration. Semi-quantitative RT-PCR detection of HIF-1αand the expression of VEGFmRNA changes. Western Blot detection of HIF-1αand VEGF protein expression. SPSS 15.0 experimental data obtained from the application of statistical analysis software system for processing.Results: 1.MTT results showed : rapamycin inhibited proliferation of human cervical cancer Hela cells, the inhibitory rate increased with the drug concentration increased, the maximum inhibition rate of up to (51.22±2.45)% at 200ng/ml, the experimental group compared with the control group, respectively, the differences were significant, but the concentration of the adjacent inter-group comparison, 12.5ng/ml group and 25ng/ml group significantly inhibited the rate of statistical significance, the remaining groups of adjacent growth inhibition concentration as the rate of increase in drug concentration, although there is an upward trend but no statistical significance (P> 0.05). 2.RT-PCR results showed : HIF-1αmRNA and VEGFmRNA have a certain amount of expression can be reduced rapamycin were hela cervical carcinoma cells, HIF-1αand the expression of VEGFmRNA. Each group the expression of HIF-1αmRNA compared with the control group the differences were significant, low- dose group, medium-dose group the expression of HIF-1αmRNA statistically significant difference; middle-and high-dose group between the expression of HIF-1αmRNA no significant differences. VEGFmRNA low-dose group compared with the control group VEGFmRNA the difference was not statistically significant, low-, medium-dose group VEGFmRNA statistically significant difference in expression; middle-and high-dose group expressed VEGFmRNA was no significant difference.3. Western blot results showed that : in the location of 120KD and 23KD visible HIF-1αand VEGF protein bands, each dose group, respectively, with the corresponding control group showed: low-dose group of HIF-1αprotein, VEGFprotein expression respectively, compared with the corresponding control group, no statistically significant difference (P = 0.429, P = 0.321); middle-and high-dose group of HIF-1α, VEGF protein expression compared with the corresponding control group, there was a significant difference; low, medium dose group and high dose groups, respectively, compared HIF-1αprotein, VEGF protein expression differences in results are statistically significant.Conclusion: 1. Rapamycin inhibited proliferation of human cervical cancer Hela cells, even very small concentrations can also be effectively inhibited the proliferation , the largest inhibition proliferation rate at (51.22±2.45)%, inhibition with the drug concentration increase is not significantly increased, 25ng/ml dose when the drug than the maximum.2. Rapamycin can be reduced in human cervical cancer Hela cells and HIF-1αmRNA expression VEGFmRNA, drug concentration in the inhibition of the most obvious when 20ng/ml.3. Rapamycin in human cervical cancer Hela cells reduced HIF-1αand VEGF protein expression, drug concentration 20ng/ml more obvious when the inhibition, 80ng/ml inhibition is more obvious4. Rapamycin could inhibit human cervical carcinoma Hela cells the expression of HIF-1αand its downstream gene reduced the expression of VEGF to inhibit tumor angiogenesis and thus tumor growth control, cancer treatment may become another new idea.