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目的探讨白头翁皂苷D联合索拉非尼对人肝癌细胞株侵袭和转移的影响。方法以人肝癌细胞株SM M C-7721细胞为研究对象,分为白头翁皂苷D组(浓度为11.9μg/L)、索拉非尼组(浓度为2.15μmol/L)、联合组(白头翁皂苷D 11.9μg/L+索拉非尼2.15μmol/L)以及对照组(普通培养液)。通过MTT法、Transwell小室实验观察白头翁皂苷D、索拉非尼单药和联合对SMMC-7721细胞的侵袭和转移的抑制作用,并用免疫组织化学方法检测CD44V6、血管内皮生长因子C(VEGF-C)、基质金属蛋白酶MMP-9基因蛋白表达的影响。结果 MTT结果显示,白头翁皂苷D、索拉非尼单药和联合对SMMC-7721细胞的增殖均有抑制作用,且24 h的抑制率都<15%;Transw ell小室实验结果显示,联合组的黏附抑制率与单药组相比,具有协同抑制作用,但未呈现时间依赖关系;联合组的侵袭抑制率和迁移抑制率均显著高于单药组(P<0.01);免疫组织化学结果表明,联合组在下调CD44V6、VEGF-C、MMP-9方面均高于单药组(P<0.05)。结论白头翁皂苷D和索拉非尼联合可协同抑制SM M C-7721细胞侵袭和转移,联合效果整体优于单药。
Objective To investigate the effect of Pulsatilla saponin D combined with sorafenib on invasion and metastasis of human hepatoma cell line. Methods Human hepatocellular carcinoma cell line SM M C-7721 cells were divided into three groups: Pulsatilla saponin group D (concentration 11.9μg / L), sorafenib group (concentration 2.15μmol / L), combination group (Pulsatilla saponin D 11.9μg / L + sorafenib 2.15μmol / L) and control group (normal medium). The inhibitory effects of Pulsatilla saponin D, sorafenib monotherapy and combined treatment on the invasion and metastasis of SMMC-7721 cells were observed by MTT assay and Transwell chamber assay. The expressions of CD44V6, VEGF-C ), Matrix metalloproteinase MMP-9 gene protein expression. Results MTT results showed that Pulsatilla saponin D, sorafenib monotherapy and combined inhibition of SMMC-7721 cell proliferation, and inhibition rate of 24 h <15%; Transw ell chamber results showed that the combined group of Compared with single drug group, the adhesion inhibition rate had a synergistic inhibitory effect but no time-dependent relationship. The invasion inhibition rate and migration inhibition rate in the combination group were significantly higher than those in the single drug group (P <0.01). The results of immunohistochemistry , And the combination group was higher than the single drug group in down-regulating CD44V6, VEGF-C and MMP-9 (P <0.05). Conclusion The combination of Pulsatilla saponin D and sorafenib can synergistically inhibit the invasion and metastasis of SM C-7721 cells, and the combined effect is better than single drug.