miRNA-449在胃癌发生中的表达及其作用机制

发布时间：2018-03-20 02:35

本文选题：胃癌　切入点：miRNA-449　出处：《皖南医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的：通过对miRNA-449基因和E2F1蛋白在胃癌组织中表达的研究,并分析miRNA-449的表达与临床资料的关系,探讨其在胃癌发生中的作用机制,为胃癌早期诊断、有效治疗和预后评估提供依据。方法：将收集到的31对胃癌及癌旁组织标本,提取组织总RNA,茎环法反转录成cDNA,实时荧光定量PCR技术检测niRNA-449基因表达水平；提取总蛋白,Western Blot方法检测E2F1蛋白表达水平。结果1.miRNA-449的表达：miRNA-449a在31例胃癌中相对于癌旁组织有23例(74.19%)表达下调,8例(25.81%)表达上调。miRNA-449a的相对表达水平：胃癌组为3.85±2.45,癌旁组为4.66±2.41,两者相比,miRNA-449a在胃癌中的表达低于癌旁组织,差异有统计学意义(配对t=2.516,P=0.017,P0.05)。miRNA-449b在31例胃癌中相对于癌旁组织有24例(77.42%)表达下调,7例(22.58%)表达上调。miRNA-449b的相对表达水平：胃癌组为3.66±2.54,癌旁组为4.27±2.33,两者相比,miRNA-449b在胃癌中的表达低于癌旁组织,差异有统计学意义(配对t=2.525,P=0.017,P0.05)。其中miRNA-449a和miRNA-449b在胃癌组织中有21例表达一致下调,6例表达一致上调。niRNA-449c在31例胃癌中相对于癌旁非肿瘤组织有18例(58.06%)表达下调,13例(41.94%)表达上调。miRNA-449c的相对表达水平：胃癌组为2.82±1.98,癌旁组为3.05-2.05,两者相比,miRNA-449c在胃癌组织与癌旁组织中的平均表达无统计学意义(配对t=0.679, P=0.503, P0.05). miRNA-449a/b/c在胃腺癌细胞株(BGC-823)中的表达均低于正常胃黏膜细胞株(GES-1) (P=0.003/0.017/0.005, P0.05)。2. miRNA-449a/b的表达与临床资料的关系：miRNA-449a/b的表达变化与淋巴结转移显著相关。miRNA-449a/b在18例有淋巴结转移的样本中均有17例表达下调(94.45%),13例无淋巴结转移的样本分别有6例(46.15%)和7例表达下调(53.85%),淋巴结转移组与无淋巴结转移组miRNA-449 a/b的下调比例,经Fisher确切概率法检验,差异有统计学意义(P=0.004/0.012,P0.05)。而miRNA-449a/b在胃癌中的表达变化与患者的年龄、性别、肿瘤的直径、肿瘤的浸润深度以及分化程度等无相关性。3.E2F1蛋白的表达：E2F1在17例胃癌中相对于癌旁组织有13例(76.47%)表达上调,4例(23.53%)表达下调。E2F1的相对表达水平：胃癌组为1.14±0.49,癌旁组为0.97±0.41,两者相比,E2F1在胃癌中的表达高于癌旁组织,差异有统计学意义(配对t=2.138,P=0.048,P0.05)。结论：1.在胃癌中,存在niRNA-449a/b基因的低表达和E2F1蛋白的高表达,提示miRNA-449a/b口E2F1与胃癌的发生相关。2. miRNA-449a/b在胃癌中的表达变化与淋巴结转移相关,提示miRNA-449a/b可能作为判别胃癌预后的指标之一。
[Abstract]:Objective: to study the expression of miRNA-449 gene and E2F1 protein in gastric carcinoma, and to analyze the relationship between the expression of miRNA-449 and clinical data, and to explore the mechanism of miRNA-449 expression in gastric carcinogenesis, so as to provide early diagnosis of gastric cancer. Methods: the total RNAs were extracted from 31 samples of gastric cancer and adjacent tissues, and then reversely transcribed into cDNAs by stem loop method. The expression level of niRNA-449 gene was detected by real-time fluorescence quantitative PCR. Results 1. The expression of miRNA-449 in 31 cases of gastric cancer was down-regulated in 23 cases (74.19%) of 31 cases of gastric cancer compared with adjacent tissues.) the expression of miRNA-449a was up-regulated in 8 cases (25.81%): the expression level of miRNA-449a was 3.85 卤2.45 in gastric cancer group and 3.85 卤2.45 in adjacent gastric carcinoma group. The expression of miRNA-449a in gastric cancer was lower than that in paracancerous tissues. The down-regulated expression of miRNA-449b in 31 cases of gastric cancer was significantly lower than that of adjacent tissues (77.42%).) the expression level of miRNA-449b in gastric cancer group was 3.66 卤2.54 and that in paracancerous group was 4.27 卤2.33. The expression of miRNA-449b in gastric cancer group was 3.66 卤2.54 and that in paracancerous group was 4.27 卤2.33. The expression of miRNA-449b in gastric cancer was higher than that in para-cancerous tissue. Below the surrounding tissue, The expression of miRNA-449a and miRNA-449b were down-regulated in 21 cases of gastric carcinoma and 6 cases were up-regulated. NiRNA-449c was down-regulated in 13 cases (41.94%) in 31 cases of gastric cancer as compared with 18 cases of non-cancerous tissues adjacent to cancer. Upregulation of the relative expression level of .miRNA-449c: 2.82 卤1.98 in gastric cancer group and 3.05-2.05 in paracancerous group. There was no significant difference in the average expression of miRNA-449c between gastric cancer tissue and paracancerous tissue (pairwise t0.679, P0. 503, P0. 05). The expression of miRNA-449a/b/c in gastric adenocarcinoma cell line BGC-823 was lower than that in gastric cancer cell line BGC-823. Normal gastric mucosal cell line (GES-1) P0. 003 / 0.017 / 0.005, P0. 05 / 0.2.The relationship between the expression of miRNA-449a/b and clinical data is significant correlation between the expression of miRNA-449a / b and lymph node metastasis. MiRNA-449a / b is down-regulated in 17 out of 18 samples with lymph node metastasis, and there are 13 cases without lymph node metastasis. The down-regulation of miRNA-449 a / b in lymph node metastasis group and non-lymph node metastasis group was 53.85% and 53.85% respectively, and the down-regulation ratio of miRNA-449 a / b in lymph node metastasis group and non-lymph node metastasis group was lower than that in non-lymph node metastasis group. By Fisher's exact probability test, the difference was statistically significant. The expression of miRNA-449a/b in gastric cancer was correlated with the age, sex and diameter of the tumor. (3. Expression of E2F1 protein in 17 cases of gastric carcinoma: 13 cases in relation to adjacent tissues.) upregulation and down-regulation of expression of E2F1: 1.14 卤0.49 in gastric cancer group and 23.53% in paracancerous tissue. The expression of E2F1 in gastric cancer was higher than that in paracancerous tissues. Conclusion: 1. There is a low expression of niRNA-449a/b gene and a high expression of E2F1 protein in gastric carcinoma, suggesting that the expression of E2F1 in miRNA-449a/b mouth is related to carcinogenesis of gastric cancer .2.2.The expression of miRNA-449a/b in gastric carcinoma is related to lymph node metastasis. The results suggest that miRNA-449a/b may be one of the predictors of gastric cancer prognosis.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.2

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