SP17、SPAG9、OY-TES-1基因在卵巢上皮性癌中的实时定量PCR表达及甲基化情况的研究

发布时间：2018-05-20 03:42

本文选题：SP17 + SPAG9　；参考：《内蒙古医科大学》2017年硕士论文

【摘要】：目的检测SP17、SPAG9、OY-TES-1这三种癌-睾丸抗原基因在卵巢上皮性癌与非卵巢癌中的q Realtime-PCR扩增基因表达及这三种基因在启动子区域的甲基化情况,探讨其与卵巢上皮性癌的关系。方法收集2015年1月至2016年4月于内蒙古自治区人民医院行卵巢手术患者的新鲜卵巢组织共60例,其中有30例新鲜的卵巢上皮性癌组织标本,及30例新鲜的非卵巢癌组织标本。q Realtime-PCR实验:从卵巢组织中提取RNA,反转录为c DNA,运用SYBR实时荧光定量PCR技术对上皮性卵巢癌及非卵巢癌中的特定RNA序列进行定量分析。甲基化实验:提取卵巢组织的DNA,运用DNA酸性亚硫酸盐转化、PCR扩增及PCR测序方法检测SP17、SPAG9、OY-TES-1在卵巢上皮性癌与非卵巢癌中Cp G岛的甲基化差异性表达情况。q Realtime-PCR实验数据应用2~(-△△CT)进行处理后,应用SPSS 19.0统计软件进行统计学分析,采用两独立样本t检验分析,P0.001有显著性差异,具有统计学意义。检测甲基化情况所得数据用SPSS 19.0进行卡方检验或Fisher确切概率法进行检验,P0.05有统计学意义。结果1.60例实验标本中,最终上皮性卵巢癌及非卵巢癌组织标本中各有25例结果在有效分析范围内;2.SP17基因的RNA在卵巢上皮性癌中的表达高于其在非卵巢癌中的表达,差异有统计学意义(t=16.913,P0.001);SPAG9基因的RNA在卵巢上皮性癌中的表达高于其在非卵巢癌中的表达,差异有统计学意义(t=14.312,P0.001);OY-TES-1基因的RNA在卵巢上皮性癌中的表达高于其在非卵巢癌中的表达,差异有统计学意义(t=15.976,P0.001);3.在25例上皮性卵巢癌中SP17、SPAG9、OY-TES-1基因启动子区去甲基化发生率分别为:56%(14/25)、52%(13/25)、44%(11/25);在25例非卵巢癌中SP17、SPAG9、OY-TES-1基因启动子区去甲基化发生率分别为:12%(3/25)、16%(4/25)、8%(2/25);这三种基因的去甲基化表达情况在上皮性卵巢癌与非卵巢癌中相比差异有统计学意义(P0.05);4.SP17、SPAG9、OY-TES-1基因启动子区去甲基化表达与上皮性卵巢癌患者的年龄、组织类型及分化程度无关(P0.05);SP17、SPAG9、OY-TES-1基因启动子区去甲基化表达与上皮性卵巢癌患者的淋巴结转移有关(P0.05);SP17、SPAG 9基因启动子区域去甲基化表达与上皮性卵巢癌患者的临床分期有关(P0.05),而OY-TES-1基因启动子区域去甲基化表达与其临床分期无关(P0.05)。结论1.SP17、SPAG9、OY-TES-1基因的RNA在卵巢上皮性癌中的表达高于其在非卵巢癌中的表达,联合检测SP17、SPAG9、OY-TES-1基因在上皮性卵巢癌患者中的RNA表达水平,可以提高上皮性卵巢癌诊断地灵敏性,有望成为诊断上皮性卵巢癌有效生物学标志物。2.在上皮性卵巢癌患者中SP17、SPAG9、OY-TES-1基因启动子区去甲基化表达率高于其在非卵巢癌患者中的表达;SP17、SPAG9、OY-TES-1基因启动子区在上皮性卵巢癌中激活,可能参与癌症的发生、发展及转归;3.SP17、SPAG9、OY-TES-1基因启动子区去甲基化表达与上皮性卵巢癌患者的淋巴结转移有关;SP17、SPAG9基因启动子区域去甲基化表达与临床分期有关,而OY-TES-1基因启动子区域去甲基化表达与临床分期无关;但这三种癌睾丸抗原均与上皮性卵巢癌患者的年龄、组织类型及分化程度均无关,说明这三种癌睾丸抗原在预测上皮性卵巢癌的预后及评定上皮性卵巢癌的临床分期中有一定的作用。
[Abstract]:Objective to detect the Q Realtime-PCR gene expression of three cancer testicular antigen genes of SP17, SPAG9 and OY-TES-1 in ovarian epithelial and non ovarian cancer and the methylation of these three genes in the promoter region, and to explore the relationship between them and ovarian epithelial cancer. Methods from January 2015 to April 2016 were collected from the people's medicine in the Inner Mongolia Autonomous Region. There were 60 cases of fresh ovarian tissue in the patients undergoing ovarian surgery, including 30 fresh ovarian epithelial tissue specimens and 30 fresh non ovarian cancer tissue specimens.Q Realtime-PCR experiment: extracting RNA from the ovarian tissue and reverse transcription of C DNA, and using SYBR real-time fluorescent quantitative PCR technique to treat epithelial ovarian and non ovarian cancer. Quantitative analysis of RNA sequence. Methylation experiment: extracting DNA of ovarian tissue, using DNA acid sulfite transformation, PCR amplification and PCR sequencing to detect SP17, SPAG9, OY-TES-1 in ovarian epithelial cancer and non ovarian cancer, Cp G Isle,.Q Realtime-PCR experimental data were treated with.Q Realtime-PCR experimental data. Statistical analysis was carried out with SPSS 19 statistical software, and two independent sample t tests were used to analyze the statistical significance. The data obtained by P0.001 were statistically significant. The data obtained from the detection of methylation were tested with the chi square test or the exact probability method of Fisher, and P0.05 had statistical significance. Results in the 1.60 experimental specimens, the final epithelial nature was found. The results of 25 cases of ovarian and non ovarian cancer were found in 25 cases, and the expression of RNA in ovarian epithelial cancer was higher than that in non ovarian cancer. The difference was statistically significant (t=16.913, P0.001), and the expression of RNA in SPAG9 gene in ovarian epithelial cancer was higher than that in non ovarian cancer. The difference was statistically significant (t=14.312, P0.001); the expression of RNA in OY-TES-1 was higher than that in non ovarian cancer, and the difference was statistically significant (t=15.976, P0.001); 3. in 25 cases of epithelial ovarian cancer, the rate of demethylation of SP17, SPAG9, and OY-TES-1 genes was 56% (14/25), 52% (13/25), 44% ( 11/25): the incidence of demethylation in the promoter region of SP17, SPAG9, and OY-TES-1 genes in 25 cases of non ovarian cancer were 12% (3/25), 16% (4/25), 8% (2/25), and the difference of the demethylation of the three genes in epithelial ovarian cancer and non ovarian cancer (P0.05); 4.SP17, SPAG9, OY-TES-1 gene promoter demethylation The expression was not related to the age, type and degree of differentiation in patients with epithelial ovarian cancer (P0.05); SP17, SPAG9, OY-TES-1 gene promoter demethylation was associated with lymph node metastasis in epithelial ovarian cancer (P0.05); SP17, SPAG 9 promoter region demethylation was related to the clinical stage of epithelial ovarian cancer (P0). .05), and the expression of OY-TES-1 gene promoter region demethylation is not related to its clinical stage (P0.05). Conclusion the expression of RNA in 1.SP17, SPAG9, and OY-TES-1 genes in ovarian epithelial cancer is higher than that in non ovarian cancer. The RNA expression of SP17, SPAG9, OY-TES-1 genes in patients with epithelial ovarian cancer can be improved. The diagnostic sensitivity of cutaneous ovarian cancer is expected to be an effective biomarker for the diagnosis of epithelial ovarian cancer,.2. in the patients with epithelial ovarian cancer, the expression rate of SP17, SPAG9, OY-TES-1 gene promoter demethylation is higher than that in non ovarian cancer patients; SP17, SPAG9, OY-TES-1 gene promoter region is activated in epithelial ovarian cancer. Can participate in the occurrence, development and prognosis of cancer; 3.SP17, SPAG9, OY-TES-1 gene promoter demethylation is related to lymph node metastasis in patients with epithelial ovarian cancer; SP17, SPAG9 gene promoter region demethylation expression is related to clinical staging, and the expression of OY-TES-1 gene promoter region demethylation is not related to clinical staging. The three kinds of testicular antigens were not related to the age, type and degree of differentiation of the patients with epithelial ovarian cancer, which showed that these three kinds of cancer testis have a certain role in predicting the prognosis of epithelial ovarian cancer and assessing the clinical stage of epithelial ovarian cancer.
【学位授予单位】：内蒙古医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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