叶酸缺乏与miR-203异常表达在宫颈癌发生中的相关性研究

发布时间：2018-05-28 04:10

本文选题：宫颈癌 + 叶酸　；参考：《山西医科大学》2014年硕士论文

【摘要】：目的：宫颈癌是女性最常见的妇科恶性肿瘤之,发病率呈逐年上升趋势。高危型人乳头瘤病毒(HR-HPV)感染是宫颈癌发生的重要病因,但并非唯一因素,故寻找HR-HPV感染的协同因素在阻断ClN进展为宫颈癌的过程中非常重要。宫颈癌的发生、发展是多因素、多步骤、长时间作用的结果,是一个及其复杂的生物学过程。目前最新研究发现,microRNA的异常表达及叶酸缺乏与宫颈癌的发生密切相关,而其具体机制尚不清楚。故本研究的目的在于：1.检测宫颈病变组织中miR-203的表达状态,探讨其在宫颈病变发生发展中的作用。2.体外不同浓度叶酸干预下检测宫颈癌Caski细胞中mi R-203的甲基化状态、mi R-203的表达水平,探讨叶酸缺乏致宫颈癌发生的可能机制。方法：1.研究对象选取2012年1月至2013年1月于我院妇科门诊就诊行阴道镜检查的90例患者,根据阴道镜宫颈活检病理诊断结果分为子宫颈上皮内瘤变(CIN)30例,鳞状细胞癌(SCC)30例作为研究组;宫颈组织病理学结果正常30例作为对照组。2.取患者的宫颈组织,RT-PCR技术检测90例患者宫颈组织中mi R-203的表达水平。3.常规培养宫颈癌Caski细胞株,取对数生长期的细胞,给予不同浓度的叶酸干预后,采用甲基化特异性PCR(MSP)法检测不同叶酸浓度组宫颈癌细胞中miR-203的甲基化状态,RT-PCR法检测不同叶酸浓度组宫颈癌细胞中miR-203的表达水平。4.采用SPSS 16.0统计软件进行统计学分析,定量资料的多组间比较采用单因素方差分析,组间的多重比较采用LSD-t检验,率的比较采用X2检验,相关性分析采用Spearman秩相关,以P0.05为差异有统计学意义。结果：1.正常宫颈组织、CIN组织、宫颈癌组织中miR-203的相对表达量分别为1.00±0.10,0.13±0.03,0.27±0.05。各组miR-203的表达量比较采用单因素方差分析,miR-203的表达量差异有统计学意义(F=6.414,P=0.005)。各组miR-203的表达量的组间比较采用LSD-t检验,正常组与CIN组间miR-203的表达量差别有统计学意义(P=0.001),正常组与宫颈癌组、CIN组与宫颈癌组miR-203的表达差别无统计学意义(P=0.151,P=0.060)。2.不同叶酸组宫颈癌细胞中,随着叶酸水平的升高,mi R-203的CpG岛甲基化率呈下降趋势(X2=27.473, P0.001), miR-203的表达水平呈上升趋势(F=427.416,P0.001)。对miR-203的CpG岛甲基化率与miR-203的表达水平进行相关性分析,呈负相关(rs=-0.886,P=0.019)。结论：1.在宫颈病变组织中较宫颈正常组织,miR-203的表达水平下调,miR-203表达水平的下调可能参与了宫颈癌的发生。2.叶酸缺乏可能会通过导致miR-203高甲基化,使其表达水平下调,进而在宫颈癌变过程中发挥一定作用。
[Abstract]:Objective: cervical cancer is the most common gynecologic malignant tumor in women, the incidence is increasing year by year. High-risk human papillomavirus (HR-HPV) infection is an important cause of cervical cancer, but it is not the only factor, so it is very important to search for co-factors of HR-HPV infection in the process of blocking the progression of ClN to cervical cancer. The occurrence and development of cervical cancer is the result of multi-factor, multi-step and long-term action, which is an extremely complicated biological process. Recent studies have found that the abnormal expression of microRNA and folic acid deficiency are closely related to the occurrence of cervical cancer, but its mechanism is not clear. Therefore, the purpose of this study is to 1: 1. To detect the expression of miR-203 in cervical lesions and to explore its role in the occurrence and development of cervical lesions. The methylation status of mi R-203 in cervical cancer Caski cells was detected by different concentrations of folic acid in vitro, and the possible mechanism of carcinogenesis induced by folic acid deficiency was discussed. Method 1: 1. From January 2012 to January 2013, 90 patients underwent colposcopy examination in gynecological outpatient clinic of our hospital. According to the pathological diagnosis results of colposcopy cervical biopsy, 30 cases of cervical intraepithelial neoplasia and 30 cases of squamous cell carcinoma (SCC) were divided into two groups: 30 cases of cervical intraepithelial neoplasia and 30 cases of squamous cell carcinoma (SCC). Cervical histopathological results were normal in 30 cases as control group. 2. 2. The expression level of mi R-203 was detected by RT-PCR in cervical tissues of 90 patients. Caski cell lines of cervical cancer were cultured routinely. The cells in logarithmic growth period were treated with folic acid at different concentrations. The methylation status of miR-203 in cervical cancer cells with different folic acid concentration was detected by methylation specific PCR. RT-PCR was used to detect the expression level of miR-203 in cervical cancer cells with different folic acid concentration. Statistical software SPSS 16.0 was used for statistical analysis. The multivariate analysis of variance was used in the comparison of quantitative data, the LSD-t test was used for the multiple comparison of the quantitative data, the X2 test was used for the rate comparison, and the Spearman rank correlation was used for the correlation analysis. P0.05 as the difference was statistically significant. The result is 1: 1. The relative expression of miR-203 in normal cervical tissues and cervical carcinoma tissues was 1.00 卤0.10 卤0.13 卤0.03 卤0.27 卤0.05, respectively. The expression of miR-203 in each group was compared by univariate analysis of variance (ANOVA). LSD-t test was used to compare the expression of miR-203 in each group. There was significant difference in the expression of miR-203 between normal group and CIN group. There was no significant difference in miR-203 expression between normal group and cervical carcinoma group (cin group and cervical cancer group). There was no significant difference in miR-203 expression between normal group and cervical cancer group. With the increase of folic acid level, the methylation rate of CpG islands in different folic acid groups decreased with the increase of folic acid level in cervical cancer cells. The expression level of miR-203 was increased with the increase of the level of folic acid in the cervical cancer cells (P _ (0.001) and X _ (2) ~ (2) O _ (27.416) P _ (0.001) (P _ (0.001). The correlation between CpG island methylation rate and miR-203 expression in miR-203 was negatively correlated with that of miR-203. Conclusion 1. The down-regulation of miR-203 expression level in cervical lesions may be involved in the occurrence of cervical cancer. Folic acid deficiency may play a role in the process of cervical cancer by inducing hypermethylation of miR-203 and down-regulation of its expression.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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