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目的:探讨左金丸通过抑制Wnt/β-catenin信号通路抗幽门螺杆菌(Helicobacter pylori,H.pylori)感染的人胃癌MKN45细胞转移的机制。方法:以H.pylori感染人胃癌MKN45细胞,采用CCK-8检测不同浓度左金丸醇提物对H.pylori感染的MKN45细胞生长抑制作用。Transwell检测低、中、高剂量(25、50、100μg/ml)左金丸对H.pylori感染的MKN45细胞转移能力的影响;Western blot检测其对H.pylori感染的MKN45细胞核中的β-catenin蛋白表达;ELISA检测其对H.pylori感染的MKN45细胞培养上清液中MMP-7表达。结果:左金丸醇提物对H.pylori感染的MKN45细胞生长具有抑制作用,呈剂量依赖性。Transwell结果显示,与正常对照组比较,H.pylori感染后MKN45细胞转移能力显著增强(P<0.01);与H.pylori感染组比较,低、中、高剂量左金丸均能够明显抑制H.pylori诱导的MKN45细胞转移能力(P<0.01),呈剂量依赖性,且左金丸能够明显下调H.pylori诱导的β-catenin在细胞核的累积及其下游靶基因MMP-7蛋白表达(P<0.01)。结论:左金丸防治H.pylori诱发胃癌细胞转移的机制可能与通过Wnt/β-catenin信号通路抑制MMP-7表达有关。
OBJECTIVE: To investigate the mechanism of Zuojin pill in inhibiting the metastasis of human gastric cancer MKN45 cells by inhibiting the Wnt / β-catenin signaling pathway against Helicobacter pylori (H. pylori) infection. Methods: Human gastric cancer MKN45 cells were infected with H.pylori and the growth inhibition of H.pylori-infected MKN45 cells was detected by CCK-8 with different concentrations of Zuojin pills. The effects of low, medium and high doses (25,50,100μg / ml) of Zuojin Pill on the metastasis of H.pylori-infected MKN45 cells were detected by Transwell assay. The expression ofβ-catenin protein in MKN45 cells infected by H.pylori The expression of MMP-7 in culture supernatant of MKN45 cells infected with H.pylori was detected by ELISA. Results: Alcohol extract of Zuojintue inhibited the growth of H.pylori-infected MKN45 cells in a dose-dependent manner. Transwell results showed that, compared with the control group, the metastasis ability of MKN45 cells was significantly increased after H.pylori infection (P <0.01). Compared with H.pylori infection group, low, medium and high doses of Zujin pills significantly inhibited H.pylori (P <0.01) in a dose-dependent manner. Zuojin Pills could significantly down-regulate the accumulation of β-catenin in nuclei and the expression of MMP-7 protein (P <0.01) . Conclusion: The mechanism of Zuo Jin Wan in preventing and treating H. pylori-induced gastric cancer cell metastasis may be related to inhibiting the expression of MMP-7 through Wnt / β-catenin signaling pathway.