Objective To investigate the effects of saponins from Schizocapsa plantaginea Hance (SFSP) on the proliferation, migration and apoptosis of human hepatocellular carcinoma SMMC-7721 cells and its toxicity to normal human hepatocytes HL-7702. Methods We chose the SMMC-7721 cells in the logarithmic phase and divided them into the administration group and the control group, cells in the administration group were treated with different concentrations of SFSP (0, 0.625, 1.25, 2.5 and 5 μg/mL) for 24 h. MTT assay was used to detect the proliferation inhibition rate of SMMC-7721 cells in order to finally calculate its half inhibitory concentration (IC50). SMMC-7721 cells in the logarithmic phase were treated with final concentrations of SFSP (0, 2, 4, and 6 μg/mL) for 24 h, and 48 h and then the scratch assay was used to observe the changes of cell migration ability. Furthermore, SMMC-7721 cells in the logarithmic phase were treated with final concentrations of SFSP (0, 2, 4, and 6 μg/mL) for 24 h, and meanwhile we observed their apoptosis and morphology by AO-EB double staining and immunofluorescence. We chose the HL-7702 cells in the logarithmic phase and separately treated them with different concentrations of SFSP (0, 2.5, 12.5, 25, 50 and 100 μg/mL) for 24 h. MTT assay was used to detect the proliferation inhibition rate of HL-7702 cells in order to finally calculate its IC50. Results After 0.625, 1.25, 2.5 and 5 μg/mL SFSP were applied to SMMC-7721 cells for 24, 48 and 72 h, the proliferation of cells was inhibited to different degrees. The inhibitory effect of SFSP on SMMC-7721 cells was in a time-and dose-dependent manner (all P<0.05). The IC50 values of SFSP at 24, 48 and 72 h were 3.75, 1.56 and 0.95 μg/mL, respectively. After 0, 2, 4, 6 μg/mL SFSP were applied to SMMC-7721 cells, the healing rate of the cells was gradually decreased and the apoptotic cells were gradually increased (all P<0.05). SMMC-7721 cells shrank, appeared round or spindle shape, tubulin β fluorescence intensity decreased and highly aggregated, and a large number of cells in the split cycle. After 0, 2.5, 12.5, 25, 50 and 100 μg/mL SFSP were applied to HL-7702 cells for 24 h, the proliferation of cells was inhibited to different degrees. The IC50 of SFSP was 47.6 μg/mL at 24 h, which was lower than that (3.75 μg/mL) of SMMC-7721 cells. Conclusions SFSP could significantly inhibit the proliferation and migration, induce the apoptosis, and block the cell cycle in human hepatocellular carcinoma SMMC-7721 cells. Meanwhile, SFSP have less toxic effect on normal human hepatocytes.%目的 观察裂果薯总皂苷(SFSP)对人肝癌细胞株SMMC-7721增殖、迁移、凋亡的影响及对人正常肝细胞HL-7702的毒性作用.方法 取对数生长期的SMMC-7721细胞,分为给药组和对照组(不加SFSP),给药组分别加入0.625、1.25、2.5、5 μg/mL的SFSP培养24 h,采用MTT法观察并计算细胞增殖抑制率和IC50;分别以0、2、4、6 μg/mL的SFSP作用于SMMC-7721细胞,分别于培养24、48 h采用划痕实验观察细胞迁移能力变化;分别以0、2、4、6 μg/mL的SFSP作用于SMMC-7721细胞,培养24 h后采用吖啶橙/溴乙双荧光染色法观察SMMC-7721细胞凋亡情况,免疫荧光法观察细胞形态.取对数生长期的HL-7702细胞,分别加入0、2.5、12.5、25、50、100 μg/mL的SFSP,测算细胞增殖抑制率及IC50.结果 0.625、1.25、2.5、5 μg/mL的SFSP作用于SMMC-7721细胞24、48、72 h后,细胞增殖受到不同程度的抑制,SFSP对SMMC-7721细胞的增殖抑制作用呈现时间与剂量依赖性(P均<0.05);SFSP在24、48、72 h时的IC50值分别为3.75、1.56、0.95 μg/mL.0、2、4、6 μg/ mL的SFSP作用于SMMC-7721细胞后,细胞划痕愈合率逐渐减小,凋亡细胞逐渐增多(P均<0.05).SFSP作用后,SMMC-7721细胞出现皱缩,呈圆形或纺锤形,微管蛋白β荧光强度降低并高度聚集,观察到大量阻滞于分裂周期中的细胞.0、2.5、12.5、25、50、100 μg/mL的SFSP作用于HL-7702细胞后,细胞增殖受到不同程度的抑制,SFSP作用24 h的IC50为47.6 μg/mL,低于作用于SMMC-7721细胞24 h时的IC50(3.75 μg/mL).结论 SFSP可抑制SMMC-7721细胞增殖与迁移,诱导其凋亡,阻滞其细胞周期,且对正常肝细胞毒性作用较小.
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