健脾消癌方对缺氧微环境诱导的结肠癌细胞生物学行为影响及抑癌机制研究

Effect and Anti-cancer Mechanism of Jianpi Xiaoai Formula on the Biological Behavior of Colon Cancer Cells in Hypoxic Micro-environment

目的研究健脾消癌方对缺氧微环境下结肠癌细胞生物学行为的影响,并探讨其抑癌机制。方法将对数生长期的结肠癌SW620细胞随机分为健脾消癌方组(10%健脾消癌方的含药血清培养)和对照组(10%的空白血清培养),两组同时在缺氧环境下诱导培养14 d。采用Real-time PCR和Western blot检测两组结肠癌细胞低氧诱导因子-1α(HIF-1α)、VEGF、Bcl-2、Caspase-3、MMP-9、Survivin mRNA及蛋白的表达;MTT法检测细胞增殖;Transwell检测细胞迁移能力;流式细胞术检测细胞凋亡。结果与对照组比较,健脾消癌方组结肠癌SW620细胞HIF-1α、VEGF、Bcl-2 mRNA和蛋白水平降低(P<0.05),Caspase-3 mRNA和蛋白水平升高(P<0.05);健脾消癌方组结肠癌SW620细胞在48、72、96 h时细胞存活率降低(P<0.01),迁移和穿透室膜细胞数降低(P<0.05),细胞凋亡率升高(P<0.05)。结论健脾消癌方能下调缺氧微环境中结肠癌细胞HIF-1α水平,间接影响其下游靶基因的表达,抑制结肠癌细胞增殖、促进细胞凋亡、减弱细胞的迁移,发挥抗癌作用。

Objective To investigate the effect of Jianpi Xiaoai Formula on the biological behavior of colon cancer cells in hypoxic microenvironment and to explore its possible mechanism.Methods SW620 colon cancer cells in logarithmic growth phase were randomly divided into the Jianpi Xiaoai Formula group(cultured with 10%Jianpi Xiaoai Formula serum)and the control group(cultured with 10%blank serum),both groups were induced and cultured in hypoxic environment for 14 days at the same time.Real-time PCR and Western blot were used to detect the expressions of hypoxia inducible factor-1α(HIF-1α),VEGF,Bcl-2,Caspase-3,MMP-9,Survivin mRNA in colon cancer cells of the two groups;MTT was used to detect cell proliferation;transwell was used to detect cell migration and invasion ability;flow cytometry was used to detect cell apoptosis.Results Compared with the control group,the mRNA and protein levels of HIF-1α,VEGF and Bcl-2 in SW620 colon cancer cells of the Jianpi Xiaoai Formula group decreased(P<0.05),Caspase-3 increased(P<0.05);the survival rate of SW620 colon cancer cells decreased at 48,72 and 96 hours(P<0.01),the number of migrating and penetrating ependymal cells decreased(P<0.05),and the cells apoptotic rate increased(P<0.05)in the Jianpi Xiaoai Formula group.Conclusion Jianpi Xiaoai Formula can down-regulate the expression of HIF-1αin colon cancer cells in hypoxic microenvironment,indirectly affect the expression of its downstream target genes,inhibit the proliferation of colon cancer cells,promote cell apoptosis,reduce cell migration,so as to play an anti-cancer role.