【摘要】 目的探讨微小RNA(miR)-200c靶向抑制肝配蛋白A1(EFNA1)基因对胃癌细胞增殖、凋亡及侵袭的影响。方法选取胃癌细胞株SGC7901,随机分为miR-200c模拟物组、模拟物对照组、miR-200c抑制物组、抑制物对照组,分别转染miR-200c模拟物、模拟物对照寡核苷酸、miR-200c抑制物、抑制物对照寡核苷酸,采用CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,Transwell法检测细胞侵袭。采用双荧光素酶实验验证miR-200c对EFNA1基因的靶向抑制作用。取本院手术切除的胃癌和相应癌旁组织标本48例份,通过免疫组化法检测EFNA1蛋白表达,分析胃癌组织中EFNA1蛋白表达与患者性别、年龄、吸烟、饮酒、病理类型、浸润深度、淋巴结及远处转移、肿瘤部位间的关系。结果与模拟物对照组相比,miR-200c模拟物组胃癌细胞SGC7901增殖活性降低(P<0.05),总凋亡率升高(P<0.05),侵袭能力降低(P<0.05)。miR-200c模拟物组的荧光素酶活性低于模拟物对照组(P<0.05),而miR-200c抑制物组的荧光素酶活性高于抑制物对照组(P<0.05)。EFNA1蛋白在胃癌组织中表达高于癌旁组织,其高表达与胃癌淋巴结转移有关(P<0.05),与其他临床病理特征均不相关(P均>0.05)。结论胃癌组织中EFNA1基因呈高表达,miR-200c可通过靶向EFNA1基因抑制胃癌细胞增殖及侵袭,促进凋亡。更多还原

【Abstract】 Objective To evaluate the effect of microRNA( miR)-200 c on regulating cell proliferation,apoptosis and invasion by targeting ephrin A1( EFNA1) in gastric cancer cells. Methods SGC7910 cells were cultivated and randomly divided into miR-200 c mimics group and mimics control group,which were then transfected by miR-200 c mimics and mimics oligonucleotides. Cell proliferation was detected by CCK-8,apoptosis was determined by flow cytometry,and cell invasion was assessed using Transwell assay. The target relationship between miR-200 c and EFNA1 was measured by the dual luciferase reporter gene system. Cancer and adjacent non-cancerous tissues were obtained from 48 patients who underwent gastrectomy at Guangzhou First People’s Hospital,and the protein expression of EFNA1 was tested by immunohistochemistry. The relationship between EFNA1 expression and gender,age,smoking status,alcohol drinking,histological type,depth of invasion,lymph node and liver metastasis was assessed. Results Compared with the mimics control group,the proliferative activity of SGC7910 cells in the miR-200 c mimics group was decreased,the whole apoptosis rate was increased and the invasive ability was decreased( all P < 0. 05). The luminescence intensity was significantly decreased in miR-200 c mimic transfected cells than in mimics control cells( P < 0. 05),and the luminescence intensity was significantly increased in miR-200 c inhibitor transfected cells than in inhibitor control cells( P < 0. 05). EFNA1 was significantly up-regulated in the gastric cancer tissues as compared with that noted in the adjacent non-cancerous tissues. The high expression of EFNA1 was associated with lymph node metastasis( P < 0. 05),but not associated with other clinicopathological characteristics( all P > 0. 05). Conclusion EFNA1 gene was highly expressed in the gastric cancer. miR-200 c can inhibit gastric cancer cell proliferation and invasion,and enhance cell apoptosis by targeting EFNA1 gene.更多还原