白花蛇舌草注射液诱导SKOV-3细胞株凋亡的研究

Study on Apoptosis Effect of Hedyotic Diffusa Willd Injection on SKOV-3 Cell Line

目的研究白花蛇舌草注射液对体外培养条件下卵巢癌细胞SKOV-3细胞增殖的影响以及诱导其凋亡的情况。方法取对数生长期的SKOV-3细胞(细胞数为5×104/mL)分别接种于不同细胞培养板上,试验组每孔加入200μL不同质量浓度的白花蛇舌草注射液,空白对照组加入等体积PRMI-1640培养液孵育,采用四甲基偶氮唑盐(MTT)比色法检测吸光度(A490值)并计算SKOV-3细胞增殖抑制率;Annexin-V-FITC/PI染色细胞后,用流式细胞仪技术检测空白对照组与试验组的细胞凋亡率,并置荧光显微镜下观察不同组细胞的凋亡形态变化。结果MTT法检测结果显示,随着白花蛇舌草注射液质量浓度和作用时间的增加,SKOV-3细胞增殖抑制率均明显增大,12 mL/L的试验组培养48 h的抑制率最高,达(35.67±2.16)%;流式细胞仪检测显示,白花蛇舌草注射液处理后可引起SKOV-3细胞凋亡率随着白花蛇舌草注射液质量浓度的增加呈升高趋势,且荧光镜下试验组细胞可呈现典型凋亡改变。结论白花蛇舌草注射液能诱导卵巢癌SKOV-3细胞凋亡。

Objective To explore the influence and apoptosis- inducing effect of Hedyotic Diffusa Willd Injection on the proliferation of human ovarian cancer cell line（SKOV-3） in vitro. Methods SKOV-3 cells（5 x 104/mL） in logarithmic growth phase were respec- tively inoculated on the different cell culture plates. The test group was added 200 IxL different concentrations of Hedyotic Diffusa Willd Injection and the control group was added isovolumetric PRMI- 1640 culture solution for incubation. The absorbance（A490） was measured with the MTT colorimetrie method and the inhibition rate of SKOV- 3 cells proliferation was calculated. After staining cells by Annexin-V- FITC/PI,the rates of cell apoptosis in the test group and the blank control group were detected by the flow cytometry and the morphological changes of apoptotic ceils in different groups were investigated under fluorescent microscope. Results The detec- tion Results of MTr method showed that the inhibition rate of SKOV- 3 cells proliferation was obviously amplified with the increase of concentration and action time of Hedyotic Diffusa Willd Injection. The inhibition rate in the test group with the concentration of 12 mL/L for 48 h culture was highest and reached （35.67 ＋2. 16）%.The flow cytometry detection showed that Treating cells by Hedyotic Dif- fusa Willd Injection could cause the increasing trend of inhibition rate of SKOV-3 cells proliferation with the concentration in- crease. The classic cellular apoptosis change in the treated SKOV- 3 ceils was observed under the fluorescence microscope. Conclusion Hedyotic Diffusa Willd Injection could induce apoptosis of SKOV- 3 cells in ovarian cancer.