【摘要】 目的探讨NTSR1基因在胃癌细胞增殖侵袭中的作用及其可能的机制。方法应用免疫组织化学Envision二步法检测分析233例胃癌组织中NTSR1蛋白表达与EGFR及HER2的关系。检测胃癌细胞株NTSR1 mRNA表达,选取表达丰度较高的细胞株作为实验对象。构建干扰效率最高的shNTSR1 MKN-45细胞稳定株。采用MTT及细胞侵袭实验检测NTSR1基因沉默对胃癌细胞增殖及侵袭能力的影响。检测shNTSR1对Cancer Phenotype信号通路中19种相关基因表达的影响。结果 (1)233例胃癌NTSR1阳性表达与EGFR及HER2表达均存在显著正相关性(P<0.05),同时EGFR与HER2两者表达亦存在显著正相关性(P<0.05)。(2)NTSR1在MKN-45中的表达丰度较高。(3)成功构建NTSR1 shRNA表达载体。(4)经NTSR1 shRNA干扰,MKN-45的细胞增殖倍数明显下调(P<0.05)。(5)经NTSR1 shRNA干扰,MKN-45的侵袭转移率显著减少(P<0.05)。(6)经shRNA干扰NTSR1表达后,MKN-45细胞的Cancer Phenotype信号通路相关基因中的p27Kip1蛋白的表达水平显著上调,同时,EGFR、HER2、EpCAM及Met也呈现较明显的下调趋势。结论 NTSR1在胃癌的发生发展中起重要作用,其可能通过多维度调控EGFR/HER2信号及p27Kip1、EpCAM、Met等Cancer Phenotype信号通路相关基因来促进胃癌细胞的增殖、侵袭及转移。更多还原

【Abstract】 Objective To investigate the role of NTSR1 gene in the proliferation and invasion of gastric cancer cells and its possible mechanism. Methods The relationship between expression of NTSR1 protein and EGFR, HER2 in 233 cases of gastric cancer was analyzed by Envision two-step immunohistochemistry. The expression of NTSR1 mRNA in gastric cancer cell line was detected, and the cell line with high abundance was selected as the experimental object. MKN-45 cells with the highest interference efficiency of shNTSR1 were constructed. MTT and cell invasion assay were used to detect the effect of NTSR1 gene silencing on the proliferation and invasion of gastric cancer cells. The effect of shNTSR1 on the expression of 19 related genes in the Cancer Phenotype signaling pathway, including EGFR and HER2, was investigated. Results(1) There was a significant positive correlation between the positive expression of NTSR1 and the expression of EGFR and HER2 in 233 cases of gastric cancer(P<0.05), and there was a significantly positive correlation between EGFR and HER2 expression(P<0.05).(2) The expression abundance of NTSR1 in MKN-45 cells was high.(3) The NTSR1 shRNA expression vector was successfully constructed to construct stable cell lines.(4) The proliferation of MKN-45 cells was significantly down-regulated by NTSR1 shRNA interference(P<0.05).(5) The invasion and metastasis rates of MKN-45 were significantly decreased by NTSR1 shRNA interference(P<0.05).(6) The expression of p27 Kip1 protein in Cancer Phenotype signaling pathway of MKN-45 cells was significantly up-regulated, meanwhile, EGFR, HER2, EpCAM and Met also showed a significant downward trend by NTSR1 shRNA interference(P<0.05). Conclusion NTSR1 plays an important role in the occurrence, invasion and metastasis of gastric cancer by regulating EGFR/HER2 signaling pathway and P27Kip1, EpCAM, Met and other Cancer Phenotype signaling pathways.更多还原