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目的探讨miR-34a靶向调控去乙酰化酶1(Sirtuin 1)表达对食管鳞癌细胞侵袭、转移的影响。方法采用miRNAs靶标预测软件、Target Scan Human5.1生物在线工具预测Sirtuin 1可能是miR-34a的靶基因。将体外培养的人食管鳞癌细胞EC9706分别转染miR-34a mimic(EC9706/miR-34a组)、miR-34a scramble(EC9706/scramble组),转染48 h后收集细胞,采用Western blotting法检测Sirtuin 1蛋白表达、Transwell侵袭试验检测穿膜细胞数、细胞划痕实验检测细胞划痕愈合率。将裸鼠随机分为对照组和miR-34a组,分别经尾静脉注射转染miR-34a scramble和转染miR-34a agomir的EC9706细胞悬液,接种8周后处死裸鼠,取出双肺,计数肺转移瘤数量。结果 EC9706/miR-34a组及EC9706/scramble组细胞Sirtuin 1蛋白相对表达量分别为0.75±0.07、1.90±0.06,穿膜细胞数分别为(123.00±8.89)、(203.33±5.86)个,细胞划痕愈合率分别为(33.67±2.08)%、(64.00±2.65)%,两组比较P均<0.01。miR-34a组和对照组裸鼠形成肺转移瘤分别为(4.67±0.82)、(8.83±1.17)个,两组比较P<0.01。结论 Sirtuin 1是miR-34a的靶基因。miR-34a通过靶向抑制Sirtuin 1蛋白表达抑制食管鳞癌EC970细胞的侵袭、转移。
Objective To investigate the effect of miR-34a targeting Sirtuin-1 expression on the invasion and metastasis of esophageal squamous cell carcinoma. Methods Using miRNAs target prediction software, Target Scan Human5.1 biological online tool predicted that Sirtuin 1 may be the target gene of miR-34a. The human esophageal squamous carcinoma cell line EC9706 was transfected with miR-34a mimic (EC9706 / miR-34a) and miR-34a scramble (EC9706 / scramble) respectively. After 48 h of transfection, cells were harvested and analyzed by Western blotting Sirtuin 1 protein expression, Transwell invasion assay to detect the number of transmembrane cells, cell scratch test to detect cell wound healing rate. The nude mice were randomly divided into control group and miR-34a group. The EC9706 cells transfected with miR-34a scramble and miR-34a agomir were transplanted via caudal vein. After being inoculated for 8 weeks, the nude mice were killed and the lungs were removed. Count the number of lung metastases. Results The relative expression levels of Sirtuin 1 protein in EC9706 / miR-34a and EC9706 / scramble groups were (0.75 ± 0.07), (1.90 ± 0.06) and (123.00 ± 8.89) and (203.33 ± 5.86) The healing rates were (33.67 ± 2.08)% and (64.00 ± 2.65)% respectively, P <0.01 in both groups. The lung metastases of miR-34a group and control group were (4.67 ± 0.82) and (8.83 ± 1.17) respectively, P <0.01 compared with the two groups. Conclusion Sirtuin 1 is the target gene of miR-34a. miR-34a inhibits the invasion and metastasis of esophageal squamous cell carcinoma EC970 cells by targeting the inhibition of Sirtuin-1 protein expression.