长链非编码基因MALAT1在乳腺癌组织中的表达及其甲基化情况的研究

发布时间：2018-05-27 04:45

本文选题：MALAT1 + 甲基化　；参考：《昆明医科大学》2015年硕士论文

【摘要】：目的：通过实时荧光定量RT-PCR(Real Time-PCR, RT-PCR)技术,研究在乳腺癌及癌旁正常乳腺组织中MALAT1基因的表达情况,经甲基化特异性PCR(Methylmion Specific PCR, MSP)对乳腺癌和癌旁正常乳腺组织两组间MALAT1基因DNA甲基化的表达差异进行检测,对MALAT1基因DNA甲基化在乳腺癌中的表达进行分析研究,探讨MALAT1基因在乳腺癌中的表达与甲基化的关系及相关临床意义。方法：采集乳腺癌组织及癌旁正常乳腺组织30对,所采集癌旁正常乳腺组织距离原发恶性肿瘤组织应该至少大于50mm,并且在同侧乳腺不同象限取材。分别提取总RNA,进行逆转录,对MALAT1在两组中的表达水平行RT-PCR方法定量,检定乳腺癌组织、癌旁正常乳腺组织中,MALAT1基因的表达情况：选用离心柱法采集乳腺癌组织、癌旁正常乳腺组织的基因组DNA,对乳腺癌组织和癌旁正常乳腺组织使用MSP法检查MALAT1的DNA甲基化情况；使用SPSS17.0统计软件对实验数据进行处理,结果行统计学分析。结果：1.在30例配对乳腺癌组织及癌旁正常乳腺组织中,MALAT1在乳腺癌组织中的表达(6.56±2.64)显著较癌旁正常标本(4.84±3.82)高,有显著统计学差异(P=0.046)。2.30例乳腺癌组织中出现甲基化5例,未甲基化25例；30例癌旁正常乳腺组织中出现甲基化12例,未甲基化18例,具有显著统计学差异(P=0.04)。结论：1.乳腺癌组织中MALAT1表达水平较癌旁正常乳腺组织中明显升高,差异有统计学意义,提示MALAT1在乳腺癌组织中表达上调与乳腺癌的发展有关,可能起到促进乳腺癌发展的作用。2.乳腺癌组织中MALAT1甲基化水平较癌旁正常乳腺组织中明显降低,差异有统计学意义,MALAT1基因启动子甲基化是导致MALAT1基因表达下调的重要机制,而且MALAT1基因启动子的异常甲基化及MALAT1基因的功能失活对乳腺癌的发展起抑制作用,MALAT1甲基化可能是一个有价值的分子标志。
[Abstract]:Objective: to study the expression of MALAT1 gene in breast cancer and adjacent normal breast tissues by real-time quantitative RT-PCR(Real Time-PCR (RT-PCR) technique. The difference of DNA methylation of MALAT1 gene between breast cancer and adjacent normal breast tissues was detected by methylation specific PCR(Methylmion Specific PCR, MSP), and the expression of MALAT1 gene DNA methylation in breast cancer was studied. To investigate the relationship between methylation and MALAT1 gene expression in breast cancer and its clinical significance. Methods: 30 pairs of breast cancer tissues and adjacent normal breast tissues were collected. The adjacent normal breast tissues should be at least more than 50 mm from the primary malignant tumor tissues, and the samples were obtained from different quadrants of the ipsilateral breast. Total RNAs were extracted, reverse transcription was performed, the expression level of MALAT1 in the two groups was quantified by RT-PCR method, and the expression of MALAT1 gene in breast cancer tissues and adjacent normal breast tissues was verified. The DNA methylation of MALAT1 in breast cancer tissues and adjacent normal breast tissues was detected by MSP method, and the experimental data were processed by SPSS17.0 software, and the results were analyzed statistically. The result is 1: 1. The expression of MALAT1 in 30 matched breast cancer tissues and adjacent normal breast tissues was significantly higher than that in adjacent normal tissues (4.84 卤3.82). There was significant difference in methylation in 5 cases of breast cancer tissues. There were 12 cases of nonmethylation and 18 cases of unmethylation in 30 cases of normal breast tissue adjacent to carcinoma. There was a significant difference between them (P < 0.04). Conclusion 1. The expression of MALAT1 in breast cancer tissues was significantly higher than that in adjacent normal breast tissues, indicating that the up-regulation of MALAT1 expression in breast cancer tissues was related to the development of breast cancer and might play a role in promoting the development of breast cancer. The level of MALAT1 methylation in breast cancer tissues was significantly lower than that in adjacent normal breast tissues, and the difference was statistically significant. The promoter methylation of MALAT1 gene was an important mechanism leading to the down-regulation of MALAT1 gene expression. The abnormal methylation of MALAT1 promoter and the functional inactivation of MALAT1 gene may inhibit the development of breast cancer. MALAT1 methylation may be a valuable molecular marker.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.9

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