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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。
Objective To investigate the expression of 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 level of kisspeptin in 50 cases of gastric adenocarcinoma and corresponding paracancerous tissues respectively. The kisspeptin small interfering RNA and control-siRNA, the kisspeptin overexpression vector (p EGFP-N1-kisspeptin) and the control empty vector (p EGFP-N1) were transfected into the gastric mucosa of MKN-45 The expression of kisspeptin, MMP-9, MMP-2, β-catenin and C-myc protein in cancer cells were detected by Western blot after 48 h. Proliferative ability, cell scratch test to detect cell migration ability. After treated with Wnt / β-catenin inhibitor FH535 in gastric adenocarcinoma cells, the cell proliferation was detected by MTT assay and the apoptosis was detected by flow cytometry. Results The expression of Kisspeptin in gastric adenocarcinoma was significantly lower than that in paracancerous tissues. Compared with the control group, kisspeptin overexpression significantly decreased the survival and migration of MKN-45 cells and the levels of MMP-9, MMP-2, β-catenin and C-myc , Knocking down the kisspeptin level, the cell survival rate and the migration rate of MMP-9, MMP-2, β-catenin, C-myc levels increased significantly. FH535 treated gastric adenocarcinoma cells proliferation and migration trend consistent with the overexpression of kisspeptin cells. Conclusions Kisspeptin is low expressed in gastric adenocarcinoma and kisspeptin inhibits the proliferation and migration of gastric adenocarcinoma through Wnt / β-catenin signaling pathway.