摘要
目的探讨基质金属蛋白酶-14(MMP14)对胃癌发生、发展的作用及其相关分子机制。方法体外培养胃癌细胞和胃黏膜上皮细胞,检测MMP14基因表达差异的统计学意义;将胃癌SGC-7901细胞体外培养分为对照组(NC-shRNA)和实验组(MMP14-shRNA),分别进行转染实验;利用四甲基偶氮唑盐(MTT)法检测细胞增殖,流式细胞技术检测细胞凋亡,黏附试验、Transwell试验分别检测细胞黏附、侵袭和迁移能力,蛋白印迹检测上皮-间质转化(EMT)和Wnt/β-catenin信号通路相关的蛋白表达。将体外转染的细胞接种于体内建立荷瘤裸鼠模型,观察和测量肿瘤体积和重量,蛋白印迹检测组织信号相关蛋白分子的表达。结果与GSE-1细胞比较,胃癌SGC-7901、BGC-823和MKN45细胞的MMP14表达均显著性上调(P<0.01);与NC-shRNA组比较,体外结果表明MMP14-shRNA能够显著抑制SGC-7901细胞的增殖、黏附、侵袭和迁移能力,影响VEGF、E-cadherin、Vimentin和Wnt/β-catenin信号蛋白的表达(P<0.05);体内结果表明,MMP14-shRNA能够显著抑制肿瘤的生长,影响Wnt/β-catenin信号蛋白的表达(P<0.05)。结论本研究表明MMP14可能通过调节Wnt/β-catenin信号通路活化影响胃癌的发生、发展。
Objective To investigate the role of matrix metalloproteinase-14(MMP14)in the development of gastric cancer and its related molecular mechanisms.Methods Gastric cancer cells and gastric mucosal epithelial cells were cultured in vitro,and the expression of MMP14 was detected by RT-PCR.The in vitro culture of gastric cancer SGC-7901 cells was divided into control group(NC-shRNA)and experiment(MMP14-shRNA),which were transfected separately.The cell proliferation was detected by MTT assay and apoptosis was detected by flow cytometry.The cell adhesion,invasion and migration ability were detected by adhesion assay and Transwell assay.Western blot detects protein expression associated with epithelial-mesenchymal transition(EMT)and Wnt/β-catenin signaling pathways.The transfected cells in vitro were seeded to establish a tumor-bearing nude mouse model in vivo.The tumor volume and weight were observed and measured,and the expression of related protein molecules was detected by Western blot.Results Compared with GSE-1 cells,the expression of MMP14 in gastric cancer SGC-7901,BGC-823 and MKN45 cells was significantly up-regulated(P<0.01).Compared with NC-shRNA group,MMP14-shRNA can significantly inhibit the ability of SGC-7901 cells to proliferate,adhere,invade and migrate as well as affect the expression of VEGF,E-cadherin,Vimentin and Wnt/β-catenin signaling proteins in vitro(P<0.05).MMP14-shRNA can inhibit the growth of tumor mass and affect the expression of Wnt/β-catenin signaling proteins in vivo(P<0.05).Conclusion This study demonstrates that MMP14 may affect the development of gastric cancer by regulating the activation of Wnt/β-catenin signaling pathway.
作者
李刚
刘江豪
罗璟璐
张新峰
Li Gang;Liu Jianghao;Luo Jinglu(Department of General Surgery,Reproductive Health Hospital of Xinjiang Uygur Autonomous Region,Xinjiang 830000,China)
出处
《医学研究杂志》
2020年第2期92-98,共7页
Journal of Medical Research
基金
新疆维吾尔自治区自然科学基金资助项目(2018D01C091)。