健脾解毒方通过miR-195抑制大肠癌细胞转移

Jianpi Jiedu Recipe inhibits metastasis of colorectal cancer cells via miR-195

目的:研究健脾解毒方通过miR-195抑制大肠癌LoVo细胞的转移能力。方法:分别将miR-195的模拟物或抑制剂转染LoVo细胞,以及采用低、中、高剂量(50、100、200μg/m L)的健脾解毒方作用LoVo细胞48h,荧光定量PCR检测细胞中miR-195表达;Transwell检测细胞转移能力;ELISA检测细胞培养上清液中MMP-2和MMP-9表达。结果:miR-195的模拟物能够显著上调其表达,抑制LoVo细胞的转移能力,以及下调细胞分泌MMP-2、MMP-9蛋白(P<0.01);而采用miR-195的抑制剂干预结果与之相反(P<0.05)。健脾解毒方能够上调LoVo细胞的miR-195表达水平(P<0.01),呈剂量依赖性。与LoVo组比较,健脾解毒方能够显著抑制LoVo细胞的转移能力,下调MMP-2、MMP-9表达(P<0.01);在转染miR-195的抑制剂的基础上再采用健脾解毒方干预,发现健脾解毒方对LoVo细胞转移能力无明显影响。结论:健脾解毒方能够抑制人肠癌细胞的转移能力,其作用机制可能与上调miR-195表达有关。

Objective: To investigate the mechanism of JianPi JieDu Recipe(JPJD) on metastasis via regulating miR-195 in colorectal cancer LoVo cells. Methods: miR-195 mimics or inhibitor was transfected into LoVo cells; JPJD in low-dose, medium-dose, and high dose(50, 100, 200 μg/m L) group treated LoVo cells for 48 hours, the expression of miR-195 was detected by real-time PCR; the migration of LoVo cells was analyzed by Transwell assay; MMP-2 and MMP-9 expression in cell culture supernatant was assayed by ELISA. Results: miR-195 mimics significantly increased miR-195 expression and down-regulated the migratory ability of LoVo cells, accompanying with the reduced expression of MMP-2 and MMP-9 in supernatant(P<0.01); on the other hand, miR-195 inhibitor had the opposite effect(P<0.01). JPJD could increase miR-195 expression(P<0.01) in dose-dependent manner. Compared with LoVo group, JPJD could inhibit the migratory ability of LoVo cells and down-regulate the expression of MMP-2 and MMP-9(P<0.01). With miR-195 inhibitor transfected into LoVo cells, JPJD had no effect on the migratory ability. Conclusion: JPJD could inhibit the migration of colorectal cancer through miR-195.