过表达kisspeptin抑制MKN-45胃腺癌细胞的增殖和迁移

发布时间：2018-07-25 19:15

【摘要】：目的探讨肿瘤转移抑制因子kisspeptin(KISS-1)在胃腺癌组织中的表达及对胃腺癌细胞增殖、迁移能力的影响。方法实时定量PCR和Western blot法分别检测50例胃腺癌组织和对应的癌旁组织中kisspeptin的mRNA和蛋白水平。分别将kisspeptin小干扰RNA(kisspeptin-siRNA)和对照小RNA(control-siRNA)、kisspeptin过表达载体(p EGFP-N1-kisspeptin)和对照空载体(p EGFP-N1)转染至MKN-45胃腺癌细胞中,培养48 h后,Western blot法检测细胞中kisspeptin、基质金属蛋白酶9(MMP-9)、MMP-2、β联蛋白(β-catenin)、C-myc蛋白水平,MTT法检测细胞增殖能力,细胞划痕实验检测细胞迁移能力。用Wnt/β-catenin信号通路抑制剂FH535作用于胃腺癌细胞后,MTT法检测细胞增殖、流式细胞术检测细胞凋亡情况。结果 Kisspeptin在胃腺癌组织中的表达水平明显低于癌旁组织。与空载体组相比,过表达kisspeptin后,MKN-45细胞存活率和迁移率、MMP-9、MMP-2、β-catenin、C-myc水平均明显降低;与control-siRNA组细胞相比,敲低kisspeptin水平后,细胞存活率和迁移率、MMP-9、MMP-2、β-catenin、C-myc水平明显增加。FH535处理后的胃腺癌细胞增殖、迁移趋势与过表达kisspeptin细胞一致。结论Kisspeptin在胃腺癌组织中低表达,kisspeptin通过Wnt/β-catenin信号通路抑制胃腺癌细胞增殖迁移能力。
[Abstract]:Objective to investigate the expression of tumor metastasis inhibitor kisspeptin (KISS-1) in gastric adenocarcinoma and its effect on the proliferation and migration of gastric adenocarcinoma cells. Methods Real-time quantitative PCR and Western blot were used to detect the mRNA and protein levels of kisspeptin in 50 gastric adenocarcinoma tissues and corresponding paracancerous tissues. Kisspeptin small interfering RNA (kisspeptin-siRNA), control small RNA (control-siRNA) kisspeptin overexpression vector (p EGFP-N1-kisspeptin) and control empty vector (p EGFP-N1) were transfected into MKN-45 gastric adenocarcinoma cells, respectively. After 48 h culture, kisspeptinin, matrix metalloproteinase-9 (MMP-9) MMP-2, 尾 -catenin and C-myc protein levels were detected by Western blot assay, and cell migration was detected by cell scratch assay. Wnt/ 尾 -catenin signaling pathway inhibitor FH535 was used to detect the proliferation of gastric adenocarcinoma cells and apoptosis by flow cytometry. Results the expression of Kisspeptin in gastric adenocarcinoma was significantly lower than that in paracancerous tissues. Compared with empty vector group, the survival rate and mobility of MKN-45 cells were significantly lower than those of empty vector group, and the levels of MMP-9, MMP-2, 尾 -cateninine C-myc were significantly decreased after overexpression of kisspeptin, and the survival rate and mobility of MMP-9, MMP-2 and 尾 -cateninine C-myc were significantly increased after treatment with .FH535 compared with those in control-siRNA group. The migration trend was consistent with overexpression of kisspeptin cells. Conclusion the low expression of Kisspeptin in gastric adenocarcinoma inhibits the proliferation and migration of gastric adenocarcinoma cells through Wnt/ 尾 -catenin signaling pathway.
【作者单位】：河北医科大学第一医院病理科;河北医科大学第一医院骨科;
【基金】：河北省卫计委青年科技课题(20150642)
【分类号】：R735.2

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