Effects Of Endostatin On Apoptosis And PARP-1 And Caspase-3 Gene Expression In Endometrial Carcinoma HEC1B Cells

Objective:By detecting the effects of endostatin on the proliferation and apoptosis of endometrial carcinoma HEC1 B cells and the expressions of ADP ribose polymerase-1(PARP-1)and cysteine aspartate-specific protease-3(caspase-3)in the endometrial carcinoma HEC1 B cells,to investigate the possible mechanism of endostatin in the pathogenesis,invasion and metastasis of endometrial carcinoma,and whether the expression of PARP-1 and caspase-3 can induce the apoptosis of HEC1 B cells in endometrial carcinoma.Methods:Endometrial cancer HEC1 B cells were treated with endostatin at a final concentration of 10,50,100,200 mmol·L-1(study group),HEC1 B cells were treated with no drug-treated as a negative control group,after 48 and 72 h incubation,the proliferation of HEC1 B cells in each group was detected by Tetrazolium bromide colorimetric(MTT)method assay,and the apoptosis of HEC1 B Cell was detected by Hoechst33258 Fluorescence Staining,the expression of PARP-1 and caspase-3 in supernatant of HEC1 B cells was detected by enzyme-linked immunosorbent assay(ELISA).SPSS 18.0 software was used to analyze the experimental results.Results:1.MTT results showed,the inhibitory rates of 48 h and 72 h in the control group were(6.24±0.39)%,(5.63±0.41)%,and the inhibitory rates of 48 h and 72 h of the test group with 10,50,100,200 mmol·L-1 endostatin intervention were(6.98±0.52)%,(8.96±0.54)%,(14.36±1.02)%,(23.16±2.45)%,(24.31±2.06)%,(29.48±2.81)%,(28.16±2.13)%,(38.49±0.68)% respectively.Compared with the negative control group,the proliferation inhibition rate of HEC1 B cells increased significantly in the different culture times of the study group(P<0.01),and the inhibition rate of HEC1 B cells increased with the increasing of culture time and drug concentration(P<0.05 or P<0.01).2.Hoechst33258 Fluorescence Staining test results showed,after endostatin interfering with HEC1 B cells for 72 h,the fluorescence staining showed that compared with the negative control group,the cells in the study group appeared nuclear pyknosis,cell fragmentation,apoptotic body formation and enhanced fluorescence intensity.3.BY ELISA method,the expression of PARP-1 protein in the supernatant of HEC1 B cells which was treated with endothelin at 50,100,200 mmol·L-1 was lower than that in the negative control group,and the expression of caspase-3 protein was significantly higher than that of the negative control group(P<0.05 or P<0.01),and showed a certain concentration-dependent.Conclusion:1.Endostatin can inhibit the proliferation of HEC1 B cells and promote apoptosis.2.The proliferation inhibition rate of endometrial carcinoma cells increased gradually with the prolongation of drug application time.3.The proliferation inhibition rate of endometrial cancer cells increased with the increase of endostatin drug concentration.4.its mechanism may be related to down-regulation of PARP-1 gene expression and up-regulation of caspase-3 gene expression to induce the apoptosis of endometrial cancer.