miRNA13在裸鼠荷人结肠癌种植瘤中的作用机制研究.docxVIP

天津医科大学硕士学位论文 中文摘要 目的：研究miRNA．132在裸鼠人结肠癌种植瘤中表达和作用。 方法：①培养人结肠癌HCT-116细胞系，建立BALB／c裸鼠种植瘤模型，分为 三组，空白组；二甲基亚砜组：二甲基亚砜；抑制剂组：STAT3抑制剂WPl066+ 二甲基亚砜；②分别于合适时间点收获肿瘤，采用TaqMan miRNA Assays技术 对各组肿瘤miRNA．132表达变化进行相对定量研究；③采用实时定量PCR技术 检测种植瘤MMP．2和MMP．9分子mRNA的表达变化，蛋白免疫组化技术检测 MMP．2和MMP。9蛋白的表达。 结果：空白组的肿瘤体积与二甲基亚砜组的肿瘤体积大小差异无统计学意义 (pO．05)，抑制剂组的肿瘤体积小于以上两组的肿瘤体积，差异具有统计学意 义(p0．05)。 空白组与抑制剂组比较，miRNA。132水平差异具有统计学意义(p0．05)。 空白组肿瘤组织的MMP．2、MMP．9分子mRNA转录水平相比二甲基亚砜 组差异无统计学意义(pO．05)。抑制剂组肿瘤组织的MMP。2、MMP．9mRNA 转录水平显著低于空白组和二甲基亚砜组，差异具有统计学意义(p0．05)。 空白组肿瘤组织的MMP．2、MMP．9蛋白表达相LL--甲基亚砜组差异无统计 学意义。抑制剂组肿瘤组织的MMP一2、MMP。9蛋白表达著低于空白组和二甲基 亚砜组，差异具有统计学意义(p0．05)。 种植瘤组织中MMP．2分子mRNA转录水平、蛋白表达与MMP．9分子mRNA 转录水平、蛋白表达分别呈正相关(P0．05)；miRNA．132水平与MMP．9分子 mRNA转录水呈正相关(P0．05) 结论：miRNA-132在裸鼠人HCT-116结肠癌种植瘤生长过程中起正向调节作用。 STAT3抑制剂WPl066在抑制淋巴管生成的同时，使miRNA．132及其下游基因 MMP一9表达减低，表现为种植瘤的生长速度减慢。 关键词：人结肠癌；种植瘤；miRNA．132；MMP．2；MMP．9；STAT3：WPl066  天津医科大学硕士学位论文 Abstract Objective：To research on expression and function of miRNA一1 32 in colonic implanted carcinoma in nude mouse． Methods：①A BALB／c nude mouse model of colonic implanted carcinoma was established by using tumor cell HCT-1 1 6 and was divided into three groups randomly blank control group；DMSO group(dimethyl sulfoxide，DMSO)and inhibitor group(WPl066，the inhibitor of STAT3+DMSO)．@The tumors will be harvested， then tested by real—time PCR for relative quantitative study for miRNA一1 32；③The mRNA transcription level of MMP一2，MMP-9 was determined by real-time quantitative PCR and the expression of MMP一2，MMP一9 were tested by immunohistochemistry staining． Results：Tumor sizes of blank control group mad DMSO group had no significant difference(p0．05)．However the size of inhibitor group was significantly smaller than other two groups(p0．05)． The expression of miRNA一1 32 has significant difference between blank control group and inhibitor group(p0．05)． The mRNA transcription levels of MMP一2，MMP一9 in tumor tissues had no significant difference between blank and DMSO group(p0．05)．Howeveg there were