虎杖苷对结肠癌细胞迁移、侵袭能力的干预效果及其可能分子机制

Intervention effect of polydatin on migration and invasion abilities of colon cancerous cells and its possible molecular mechanism

目的分析虎杖苷对人结肠癌SW480细胞迁移和侵袭能力的影响,并基于上皮细胞-间充质转化(EMT)标志物及信号传导与转录激活因子3(STAT3)信号通路探讨其可能的分子机制。方法使用不同浓度(0μmol/L、25μmol/L、50μmol/L、75μmol/L、100μmol/L、150μmol/L)的虎杖苷干预SW480细胞24 h后,采用CCK-8法检测SW480细胞的增殖率,筛选后续实验的干预浓度。使用0μmol/L、25μmol/L、50μmol/L的虎杖苷干预SW480细胞(分别设为0μmol/L虎杖苷组、25μmol/L虎杖苷组、50μmol/L虎杖苷组)24 h后,采用划痕实验、Transwell实验检测SW480细胞迁移和侵袭的能力,采用Western blot检测α-平滑肌肌动蛋白(α-SMA)和波形蛋白的表达水平、磷酸化STAT3(p-STAT3)/STAT3值。结果(1)与0μmol/L虎杖苷相比,经25μmol/L、50μmol/L虎杖苷干预后SW480细胞的增殖率差异无统计学意义(P>0.05),经75μmol/L、100μmol/L、150μmol/L虎杖苷干预后SW480细胞的增殖率降低(P<0.05)。与0μmol/L虎杖苷组相比,25μmol/L虎杖苷组和50μmol/L虎杖苷组的划痕愈合率降低(P<0.05)。Transwell细胞迁移和侵袭实验结果显示,0μmol/L虎杖苷组、25μmol/L虎杖苷组、50μmol/L虎杖苷组的平均穿膜细胞数依次减少(P<0.05)。与0μmol/L虎杖苷组相比,50μmol/L虎杖苷组中α-SMA的表达水平、p-STAT3/STAT3值降低,25μmol/L虎杖苷组和50μmol/L虎杖苷组中α-SMA、波形蛋白的表达水平降低(P<0.05)。结论虎杖苷可能通过抑制STAT3信号通路的活化而逆转EMT,从而发挥抑制SW480细胞迁移和侵袭的作用。

Objective To analyze the effect of polydatin on migration and invasion abilities of human colon cancer SW480 cells,and to explore its possible mechanism based on epithelial-mensenchymal transition(EMT)marker and signal transducer and activator of tanscription 3(STAT3)signaling pathway.Methods After 24 hours of SW480 cells intervention by the application of polydatin in different concentrations(0μmol/L,25μumol/L,50μmol/L,75μumol/L,100μumol/L,and 150μmol/L),the CCK-8 method was used to detect the proliferation rate of SW480 cells for screening intervention concentration for follow-up experiment.After 24 hours of SW480 cells intervention by employing polydatin of 0μumol/L,25μumol/L,and 50μmol/L(setting as 0μmol/L polydatin group,25μmol/L polydatin group,and 50μmol/L polydatin group,respectively),the scratch test and Transwell test were used to detect migration and invasion abilities of SW480 cells.The expressions of a-smooth muscle actin(a-SMA)and vimentin,and phosphorylated STAT3(p-STAT3)/STAT3 value were detected by using the W esterm blot.Results(1)Compared with the 0μmol/L polydatin group,there was no statistically significant difference in the proliferation rate of SW480 cells between post-intervention polydatin of 25μmol/L and 50μumol/L(P>0.05);moreover,the proliferation rate of SW480 cells after intervention by polydatin of 75μmol/L,100μmol/L,and 150μmol/L were decreased(P<0.05).Compared with the 0μumol/L polydatin group,the scratch healing rates of the 25μmol/L polydatin group and the 50 pumol/L polydatin group were decreased(P<0.05).The results of Transwell cell migration and invasion tests revealed that average number of transmembrane cells was decreased successively in the 0μmol/L polydatin group,the 25μmol/L polydatin group,and the 50μmol/L polydatin group(P<0.05).Compared with the 0μumol/L polydatin group,the expression of a-SMA,and p-STAT3/STAT3 value were decreased in the 50μumol/L polydatin group,and the expressions of a-SMA and vimentin in the 25μmol/L polydatin group and the 50μmol/L polydatin group were decreased(P<0.05).Conclusion Polydatin may reverse EMT by inhibiting activation of STAT3 signaling pathway,thereby inhibiting efects of migration and invasion of SW480 cells.