MicroRNA-196a对食管癌预后的评估价值及其生物学行为的调控机制

发布时间：2018-03-09 02:29

  本文选题：微小RNA　切入点：食管癌　出处：《中国现代医学杂志》2017年13期 　论文类型：期刊论文

【摘要】：目的探讨micro RNA-196a(miR-196a)对食管癌预后的评估价值及其生物学行为的调控机制。方法收集120例行食管癌根治性手术切除的食管癌组织及癌旁组织,逆转录聚合酶链反应(RT-PCR)检测组织miR-196a表达水平,分析miR-196a表达与食管癌患者临床资料的关系。对食管癌患者进行随访,记录患者总生存期(OS)和无病生存期(DFS);以OS和DFS作为评价指标,采用单变量和多变量Cox比例风险模型评价患者预后的影响因素。分别采用miR-196a mimic、NC-mimic、miR-196a inhibitor、NC-inhibitor转染食管癌TE1细胞,MTT实验检测细胞增殖能力,Transwell实验检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力,Western blot检测细胞ANXA1、NTN4、HMGA2、HOXB8蛋白表达。结果 miR-196a在食管癌组织中表达水平高于癌旁组织(P0.05);在TE1细胞中,miR-196a mimic组miR-196a表达水平高于NC-mimic组,miR-196a inhibitor组miR-196a表达水平低于NC-inhibitor组(P0.05),证实细胞转染实验成功。将食管癌患者按照miR-196a表达分为miR-196a高表达组51例和低表达组69例,miR-196a表达与年龄、性别、分化程度、N分期、肿瘤位置等无关(P0.05),随着T分期、TNM分期和肿瘤直径增加,miR-196a高表达率升高(P0.05)。生存分析显示,miR-196a高表达患者总生存期(P=0.015)和无病生存期(P=0.017)低于miR-196a低表达者;单因素和多因素分析显示,T分期、miR-196a表达是影响患者总生存期和无病生存期的的独立危险因素(均P0.05)。MTT实验结果显示,第48~120 h,miR-196a mimic组吸光度值高于NC-mimic组,miR-196a inhibitor组吸光度值低于NC-inhibitor组(P0.05);Transwell小室实验显示,miR-196a mimic组穿膜细胞数量高于NC-mimic组,miR-196a inhibitor组穿膜细胞数量低于NC-inhibitor组(P0.05);细胞划痕实验显示,miR-196a mimic组细胞迁移距离高于NC-mimic组,miR-196a inhibitor组细胞迁移距离低于NC-inhibitor组(P0.05)。Western blot实验结果显示,miR-196a mimic组、NC-mimic组、miR-196a inhibitor组、NC-inhibitor组HMGA2、HOXB8蛋白表达比较差异均无统计学意义(P0.05),miR-196a mimic组ANXA1、NTN4蛋白表达低于NC-mimic组,miR-196a inhibitor组ANXA1、NTN4蛋白表达高于NC-inhibitor组(P0.05)。结论 miR-196a能够作为食管癌预后的预测指标之一;miR-196a可能通过抑制ANXA1、NTN4基因的表达参与调控食管癌细胞的增殖、侵袭和迁移等生物学行为。
[Abstract]:Objective to investigate the value of micro RNA-196 miR-196a in evaluating the prognosis of esophageal carcinoma and the mechanism of its biological behavior. Methods 120 cases of esophageal carcinoma tissues and adjacent tissues were collected. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of miR-196a and analyze the relationship between the expression of miR-196a and the clinical data of patients with esophageal carcinoma. Univariate and multivariate Cox proportional risk models were used to evaluate the prognostic factors of patients. The proliferation of esophageal carcinoma TE1 cells transfected with miR-196a mimicine NC-mimicum miR-196a inhibitor was measured by MTT assay and transwell assay was used to detect the invasion ability of esophageal cancer cells. Blot was used to detect the expression of ANXA1NTN4HMGA2OXB8 protein. Results the expression level of miR-196a in esophageal carcinoma tissue was higher than that in adjacent tissue P0.05A, and the miR-196a expression level in TE1 cells was higher than that in NC-mimic group. According to the expression of miR-196a, the patients with esophageal carcinoma were divided into miR-196a high expression group (51 cases) and low expression group (69 cases). Sex, degree of differentiation and N stage, tumor location were not related to P0.05, but with the increase of T stage and tumor diameter, the high expression rate of miR-196a increased (P 0.05). Survival analysis showed that the total survival time (P0.015) and disease-free survival time (P0.017) of patients with high expression of miR-196a were lower than those of patients with low expression of miR-196a. Univariate and multivariate analysis showed that the expression of miR-196a in T stage was an independent risk factor for the overall survival and disease-free survival of patients (all P0.05. The absorbance of miR-196a mimic group was higher than that of NC-mimic group inhibitor group was lower than that of NC-inhibitor group (P0.05) Transwell chamber experiment showed that the number of perforating cells in mimic group was higher than that in NC-mimic group (P 0.05), and the cell scratch test showed that the number of perforating cells in NC-mimic group was lower than that in NC-inhibitor group (P0.05), and the cell scratch test showed that the number of perforating cells in mimic group was lower than that in NC-inhibitor group (P0.05). Cell migration distance was higher than that in NC-mimic group (P 0.05). Western blot assay showed that the expression of HMGA2HXB8 protein in NC-mimic group of mimic group was lower than that in mimic group of NC-mimic group. There was no significant difference in the expression of HMGA2HOXB8 protein in NC-inhibitor group. There was no significant difference in the expression of ANXA1nTN4 protein in mimic group of NC-mimic group compared with that in mimic group of NC-mimic group (P0.05miR-196a mimic group). The expression of ANXA1nTN4 protein in mimic group was lower than that in mimic group of NC-mimic group. Conclusion the expression of ANXA1nTN4 protein is higher than that of NC-inhibitor group P0.05.Conclusion miR-196a may play an important role in regulating the proliferation of esophageal cancer cells by inhibiting the expression of ANXA1NTN4 gene. Biological behaviors such as invasion and migration.
【作者单位】： 河南科技大学第一附属医院肿瘤外科二病区;
【分类号】：R735.1
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