miR-93-5p在直肠癌组织中的表达及其临床意义
发布时间：2018-09-13 13:33
【摘要】：目的:结直肠癌是世界范围内常见的恶性肿瘤之一。我国的结直肠癌发病率和死亡率在不断上升。由于结直肠癌通常起病隐匿,主要就诊症状如便血、大便习惯改变等多与某些良性疾病相似,早期诊断较为困难,因此大部分患者确诊时已处于中晚期。越来越多的证据显示,微小RNA(miRNA)参与着细胞增殖、分化、凋亡等重要生命过程,与多种疾病密切相关,在肿瘤发生发展中类似抑癌基因或癌基因的功能。MiRNA在相关癌症患者的肿瘤组织或血液循环中存在着特异性的表达,可以用许多常规方法进行差异表达分析及定量检测,具有良好的应用前景。因此我们研究直肠癌组织中miRNA的特异性表达,并探索其是否可以作为直肠癌诊断和评估预后的预测因子。方法:收集2015年12月至2016年6月期间就诊于安徽省立医院并经病理科医师确诊后的直肠癌患者手术切除的组织,包括肿瘤组织和距离肿瘤2cm以外的周围正常黏膜组织,所有患者均未进行过任何术前放疗或化疗。取同一病例的癌组织和癌旁正常组织为1对,组织送检后立刻行福尔马林固定。所有患者的临床分期经过专科医师评判。随机选取5对组织进行基因芯片检测,初步筛选出具有差异表达的miRNA,再利用qRT-PCR的方法验证两种方法检测结果的一致性,随后筛选出1~2种最优的miRNA扩大样本量进行进一步验证。证实差异显著性后,分析miRNA表达量与临床病理参数之间的相关性,探讨其可能的临床意义。结果:经过基因芯片分析,从3100种miRNA中检测出上调的共有138种,下调的有116种。结合差异倍数和探针信号值初步筛选出5种,分别是上调的hsa-miR-93-5p,hsa-miR-20a-5p,hsa-miR-106a-5p,和下调的hsa-miR-27a-3p,hsa-miR-4707-5p。在对这5对样本进行qRT-PCR检测之后我们发现,这5种miRNA在PCR中检测出的趋势与芯片检测均一致,但hsa-miR-20a-5p(P=0.3221)与hsa-miR-4707-5p(P=0.9369)的表达量在肿瘤组织与正常组织之间差异无统计学意义。比较结果并查找相关文献后,我们最终选取进行后续实验的上调miRNA为hsa-miR-93-5p(肿瘤组织表达量是正常组织的3.165倍,P=0.0058),下调miRNA为hsa-mi R-27a-3p(肿瘤组织表达量是正常组织的0.392倍,p=0.0251)。随后的扩大样本实验中,miR-93-5p在肿瘤组织中过表达的病例数占64.79%(46/71),其在肿瘤组织与正常组织中的表达差异具有统计学意义(P0.05,n=71)。而mi R-27a-3p在肿瘤组织中呈现过表达的病例数占54.9%(39/71),肿瘤组织中的表达量高于正常组织,且差异具有统计学意义,与前期实验结果不一致,另外mi R-27a-3p的数据标准差较大,因此最终选择miR-93-5p进行临床病理参数分析,发现肿瘤组mi R-93-5p的表达量与肿瘤体积大小(P=0.004)、治疗前CEA水平(P=0.001)和淋巴结转移阳性患者其受侵淋巴结数目相关(rs=0.534,P=0.005)。结论:在直肠癌组织中存在特异性表达的miRNA。miR-93-5p在直肠癌中呈现出高表达,并与肿瘤体积,治疗前CEA水平和淋巴结转移阳性患者受侵淋巴结数目存在相关性,也许能在今后作为新型生物标记物在临床医师的诊疗过程中提供一定的指导意义。
[Abstract]:Objective:Colorectal cancer is one of the most common malignant tumors in the world.The morbidity and mortality of colorectal cancer is increasing in China.Because colorectal cancer usually has a hidden onset and its main symptoms such as blood in stool and changes in stool habits are similar to some benign diseases,early diagnosis is difficult,so most patients have been diagnosed at the time of diagnosis. More and more evidences show that microRNAs (microRNAs) are involved in cell proliferation, differentiation, apoptosis and other important life processes, closely related to a variety of diseases, similar to tumor suppressor genes or oncogene function in the development of tumors. MiRNA in the tumor tissues or blood circulation of cancer patients with specific expression, can be. Therefore, we studied the specific expression of microRNAs in rectal cancer tissues and explored whether microRNAs could be used as predictors for the diagnosis and prognosis of rectal cancer. All patients had not received any preoperative radiotherapy or chemotherapy. One pair of cancerous tissues and normal tissues adjacent to the cancer were taken from the same patient and fixed with formalin immediately after examination. Five pairs of tissues were randomly selected for microarray detection, and the results of differentially expressed microRNAs were preliminarily screened out. The consistency of the two methods was verified by qRT-PCR, and then 1-2 optimal microRNAs were selected for further validation. Results: A total of 138 kinds of up-regulated and 116 kinds of down-regulated microRNAs were detected from 3100 kinds of microRNAs by microarray analysis. Five kinds of up-regulated hsa-microRNA-93-5p, hsa-microRNA-20a-5 were preliminarily screened by combining differential multiples and probe signal values. P, hsa-microRNA-106a-5p, and down-regulated hsa-microRNA-27a-3p, hsa-microRNA-4707-5p. After qRT-PCR detection of these five pairs of samples, we found that the five microRNAs detected in PCR had the same trend as chip detection, but the expression of hsa-microRNA-20a-5p (P = 0.3221) and hsa-microRNA-4707-5p (P = 0.9369) had no significant difference between tumor tissue and normal tissue. Comparing the results and looking up the relevant literature, we finally selected the up-regulated microRNA for follow-up experiments as hsa-microRNA-93-5p (tumor tissue expression was 3.165 times of normal tissue, P = 0.0058), down-regulated microRNA as hsa-mi R-27a-3p (tumor tissue expression was 0.392 times of normal tissue, P = 0.0251). Subsequently, in the expanded sample experiment, microRNA-9 The overexpression of 3-5p in tumor tissues accounted for 64.79% (46/71), and the expression of 3-5p in tumor tissues was significantly different from that in normal tissues (P 0.05, n = 71). In addition, the standard deviation of MIR-27a-3p was large. Therefore, the expression of MIR-93-5p in tumor group was correlated with tumor size (P = 0.004), CEA level before treatment (P = 0.001) and the number of invasive lymph nodes in patients with positive lymph node metastasis (rs = 0.534, P = 0.004). CONCLUSION: There is a high expression of microRNAs. MicroRNAs-93-5p in rectal cancer tissues, which is correlated with tumor volume, CEA level before treatment and the number of invaded lymph nodes in patients with positive lymph node metastasis. It may provide a new biomarker for clinicians in the future. Definite guiding significance.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.37
，
本文编号：2241320
[Abstract]:Objective:Colorectal cancer is one of the most common malignant tumors in the world.The morbidity and mortality of colorectal cancer is increasing in China.Because colorectal cancer usually has a hidden onset and its main symptoms such as blood in stool and changes in stool habits are similar to some benign diseases,early diagnosis is difficult,so most patients have been diagnosed at the time of diagnosis. More and more evidences show that microRNAs (microRNAs) are involved in cell proliferation, differentiation, apoptosis and other important life processes, closely related to a variety of diseases, similar to tumor suppressor genes or oncogene function in the development of tumors. MiRNA in the tumor tissues or blood circulation of cancer patients with specific expression, can be. Therefore, we studied the specific expression of microRNAs in rectal cancer tissues and explored whether microRNAs could be used as predictors for the diagnosis and prognosis of rectal cancer. All patients had not received any preoperative radiotherapy or chemotherapy. One pair of cancerous tissues and normal tissues adjacent to the cancer were taken from the same patient and fixed with formalin immediately after examination. Five pairs of tissues were randomly selected for microarray detection, and the results of differentially expressed microRNAs were preliminarily screened out. The consistency of the two methods was verified by qRT-PCR, and then 1-2 optimal microRNAs were selected for further validation. Results: A total of 138 kinds of up-regulated and 116 kinds of down-regulated microRNAs were detected from 3100 kinds of microRNAs by microarray analysis. Five kinds of up-regulated hsa-microRNA-93-5p, hsa-microRNA-20a-5 were preliminarily screened by combining differential multiples and probe signal values. P, hsa-microRNA-106a-5p, and down-regulated hsa-microRNA-27a-3p, hsa-microRNA-4707-5p. After qRT-PCR detection of these five pairs of samples, we found that the five microRNAs detected in PCR had the same trend as chip detection, but the expression of hsa-microRNA-20a-5p (P = 0.3221) and hsa-microRNA-4707-5p (P = 0.9369) had no significant difference between tumor tissue and normal tissue. Comparing the results and looking up the relevant literature, we finally selected the up-regulated microRNA for follow-up experiments as hsa-microRNA-93-5p (tumor tissue expression was 3.165 times of normal tissue, P = 0.0058), down-regulated microRNA as hsa-mi R-27a-3p (tumor tissue expression was 0.392 times of normal tissue, P = 0.0251). Subsequently, in the expanded sample experiment, microRNA-9 The overexpression of 3-5p in tumor tissues accounted for 64.79% (46/71), and the expression of 3-5p in tumor tissues was significantly different from that in normal tissues (P 0.05, n = 71). In addition, the standard deviation of MIR-27a-3p was large. Therefore, the expression of MIR-93-5p in tumor group was correlated with tumor size (P = 0.004), CEA level before treatment (P = 0.001) and the number of invasive lymph nodes in patients with positive lymph node metastasis (rs = 0.534, P = 0.004). CONCLUSION: There is a high expression of microRNAs. MicroRNAs-93-5p in rectal cancer tissues, which is correlated with tumor volume, CEA level before treatment and the number of invaded lymph nodes in patients with positive lymph node metastasis. It may provide a new biomarker for clinicians in the future. Definite guiding significance.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.37
，
本文编号：2241320
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