摘要
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 investigated the mechanism of ＂Zuo Jin Pills＂ on metastasis through regulating Wnt/β-catenin signaling pathway in Helicobacter pylori （H. pylori） infected gastric cancer cell line MKN45. Methods： Human gastric MKN45 cancer cells were infected with H. pylori by co- culture, and then the effect of ＂ Zuo Jin Pills＂ alcohol extracts on the proliferation of H. pylori infected MKN45 cells was tested by CCK-8 assay. To observe the inhibitory effect of ＂Zuo Jin Pills＂ in low-dose, medium-dose, and high-dose group （25, 50, 100 μg/ml） on the migration of MKN45 cells was analyzed by Transwell assay. Expression of β-catenin expression in nuclear protein was detected by Western blot; MMP-7 expression in cell culture supernatant was assayed by ELISA. Results： ＂Zuo Jin Pills＂ alcohol extracts had obvious inhibition on the proliferation of H. pylori infected MKN45 cells in dose-dependent manner. Transwell results showed that the ability of MKN45 cells to migrate increased significantly after infected with H. pylorl （ P 〈 0.01 ） ; ＂Zun Jin Pills＂ could down- regulate the migratory ability of MKN45 cells induced by H. pylori in a dose-dependent manner compared with the H. pylori infection group （P 〈 0. 01 ）. ＂Zuo Jin Pills＂ reduced the accumulation of β- catenin in the nuclear of H. pylori infected MKN45 cells, following by the decrease of the β-catenin targeted MMP-7 expression （P 〈 0.01 ）. Conclusion： The mechanism of ＂Zuo Jin Pills＂ in preventing and treating the metastasis of H. pylori infected gastric cancer cells may be association with blocking Wnt/β-catenin signal pathway and reducing, MMP-7 expression.
出处
《上海中医药大学学报》
CAS
2015年第2期44-47,52,共5页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81273958)
上海市优秀学科带头人计划资助项目(XBR2011061)
上海市教委创新基金资助项目(13YZ045
12ZZ118)
上海市卫生局科研基金资助项目(20114Y001)
上海中医药大学预算内科研项目(2014YSN44)
