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目的:观察健脾消癌方对结肠癌HCT116细胞中微小RNA-21(microRNA-21,miR-21),第10号染色体丢失的磷酸酶基因(phasphatase and tensin homo-log deleted in chromosome 10,PTEN)表达的影响,探讨健脾消癌方抑制结肠癌转移的可能作用机制。方法:10%,15%,20%健脾消癌方含药血清处理HCTI16细胞后,实时荧光定量PCR(Real-time PCR)检测miR-21,PTEN mRNA的表达水平,蛋白免疫印迹法(Western blot)检测PTEN蛋白的表达水平。结果:与空白组比较,健脾消癌方低剂量组miR-21 mRNA相对表达量降低,PTEN mRNA相对表达量升高,但差异无统计学意义;健脾消癌方中剂量组miR-21mRNA相对表达量明显降低,PTEN mRNA相对表达量明显升高(P<0.05);健脾消癌方高剂量组miR-21 mRNA相对表达量显著降低,PTEN mRNA相对表达量显著升高(P<0.(01);与健脾消癌方中剂量组、低剂量组比较,健脾消癌方高剂量组miR-21mRNA相对表达量明显降低,健脾消癌方高剂量组PTEN mRNA相对表达量明显升高(P<0.05)。与空白组比较,健脾消癌方中剂量组、低剂量组PTEN蛋白相对表达量升高,差异无统计学意义;与空白组比较,健脾消癌方高剂量组PTEN蛋白相对表达量明显上升(P<0.05);与健脾消癌方中剂量组、低剂量组比较,健脾消癌方高剂量组PTEN蛋白相对表达量明显升高(P<0.05)。结论:健脾消癌方可能通过降低miR-21的表达,上调miR-21靶基因PTEN蛋白表达抑制结肠癌的增殖。
Objective: To observe the effects of Jianpi XiaoJian Fang on the expression of microRNA-21 (miR-21), the phosphatase gene on chromosome 10 (PTEN) in human colon cancer HCT116 cells ), And to explore the possible mechanism of Jianpi Xiaohua Decoction inhibiting colon cancer metastasis. Methods: HCTI16 cells were treated with 10%, 15%, 20% of Jianpi Xiaohua Decoction containing serum. The expression of miR-21 and PTEN mRNA was detected by real-time PCR. Western blotting was used to detect the expression of miR- blot to detect the expression of PTEN protein. Results: Compared with the blank group, the relative expression of miR-21 mRNA was decreased and the relative expression of PTEN mRNA was increased in Jianpi Xiaowan Fang low dose group, but the difference was not statistically significant; (P <0.05). The relative expression of miR-21 mRNA was significantly decreased and the relative expression of PTEN mRNA was significantly increased (P <0. 05) (01) .Compared with Jianpi Xiaowu Fang medium dose group and low dose group, the relative expression level of miR-21mRNA in high dose Jianpi Xiaoyao decoction group was significantly decreased, the relative expression level of PTEN mRNA (P <0.05) .Compared with the blank group, the relative expression of PTEN protein in Jianpi Xiaohua decoction medium dose group and low dose group increased, the difference was not statistically significant; compared with the blank group, The relative expression of PTEN protein in high dose group was significantly increased (P <0.05). Compared with the middle dose Jianpixiaohua decoction group and the low dose group, the relative expression of PTEN protein in Jianpi Xiaohua Fang high dose group was significantly increased (P < 0.05) .Conclusion: Jianpi Xiaohua decoction may inhibit the colon by decreasing the expression of miR-21 and up-regulating the expression of PTEN protein of miR-21 target gene Cancer proliferation.