本文選題：SP17 + SPAG9��； 參考：《內(nèi)蒙古醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的檢測(cè)SP17、SPAG9、OY-TES-1這三種癌-睪丸抗原基因在卵巢上皮性癌與非卵巢癌中的q Realtime-PCR擴(kuò)增基因表達(dá)及這三種基因在啟動(dòng)子區(qū)域的甲基化情況,探討其與卵巢上皮性癌的關(guān)系。方法收集2015年1月至2016年4月于內(nèi)蒙古自治區(qū)人民醫(yī)院行卵巢手術(shù)患者的新鮮卵巢組織共60例,其中有30例新鮮的卵巢上皮性癌組織標(biāo)本,及30例新鮮的非卵巢癌組織標(biāo)本。q Realtime-PCR實(shí)驗(yàn):從卵巢組織中提取RNA,反轉(zhuǎn)錄為c DNA,運(yùn)用SYBR實(shí)時(shí)熒光定量PCR技術(shù)對(duì)上皮性卵巢癌及非卵巢癌中的特定RNA序列進(jìn)行定量分析。甲基化實(shí)驗(yàn):提取卵巢組織的DNA,運(yùn)用DNA酸性亞硫酸鹽轉(zhuǎn)化、PCR擴(kuò)增及PCR測(cè)序方法檢測(cè)SP17、SPAG9、OY-TES-1在卵巢上皮性癌與非卵巢癌中Cp G島的甲基化差異性表達(dá)情況。q Realtime-PCR實(shí)驗(yàn)數(shù)據(jù)應(yīng)用2~(-△△CT)進(jìn)行處理后,應(yīng)用SPSS 19.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)學(xué)分析,采用兩獨(dú)立樣本t檢驗(yàn)分析,P0.001有顯著性差異,具有統(tǒng)計(jì)學(xué)意義。檢測(cè)甲基化情況所得數(shù)據(jù)用SPSS 19.0進(jìn)行卡方檢驗(yàn)或Fisher確切概率法進(jìn)行檢驗(yàn),P0.05有統(tǒng)計(jì)學(xué)意義。結(jié)果1.60例實(shí)驗(yàn)標(biāo)本中,最終上皮性卵巢癌及非卵巢癌組織標(biāo)本中各有25例結(jié)果在有效分析范圍內(nèi);2.SP17基因的RNA在卵巢上皮性癌中的表達(dá)高于其在非卵巢癌中的表達(dá),差異有統(tǒng)計(jì)學(xué)意義(t=16.913,P0.001);SPAG9基因的RNA在卵巢上皮性癌中的表達(dá)高于其在非卵巢癌中的表達(dá),差異有統(tǒng)計(jì)學(xué)意義(t=14.312,P0.001);OY-TES-1基因的RNA在卵巢上皮性癌中的表達(dá)高于其在非卵巢癌中的表達(dá),差異有統(tǒng)計(jì)學(xué)意義(t=15.976,P0.001);3.在25例上皮性卵巢癌中SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)去甲基化發(fā)生率分別為:56%(14/25)、52%(13/25)、44%(11/25);在25例非卵巢癌中SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)去甲基化發(fā)生率分別為:12%(3/25)、16%(4/25)、8%(2/25);這三種基因的去甲基化表達(dá)情況在上皮性卵巢癌與非卵巢癌中相比差異有統(tǒng)計(jì)學(xué)意義(P0.05);4.SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)去甲基化表達(dá)與上皮性卵巢癌患者的年齡、組織類(lèi)型及分化程度無(wú)關(guān)(P0.05);SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)去甲基化表達(dá)與上皮性卵巢癌患者的淋巴結(jié)轉(zhuǎn)移有關(guān)(P0.05);SP17、SPAG 9基因啟動(dòng)子區(qū)域去甲基化表達(dá)與上皮性卵巢癌患者的臨床分期有關(guān)(P0.05),而OY-TES-1基因啟動(dòng)子區(qū)域去甲基化表達(dá)與其臨床分期無(wú)關(guān)(P0.05)。結(jié)論1.SP17、SPAG9、OY-TES-1基因的RNA在卵巢上皮性癌中的表達(dá)高于其在非卵巢癌中的表達(dá),聯(lián)合檢測(cè)SP17、SPAG9、OY-TES-1基因在上皮性卵巢癌患者中的RNA表達(dá)水平,可以提高上皮性卵巢癌診斷地靈敏性,有望成為診斷上皮性卵巢癌有效生物學(xué)標(biāo)志物。2.在上皮性卵巢癌患者中SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)去甲基化表達(dá)率高于其在非卵巢癌患者中的表達(dá);SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)在上皮性卵巢癌中激活,可能參與癌癥的發(fā)生、發(fā)展及轉(zhuǎn)歸;3.SP17、SPAG9、OY-TES-1基因啟動(dòng)子區(qū)去甲基化表達(dá)與上皮性卵巢癌患者的淋巴結(jié)轉(zhuǎn)移有關(guān);SP17、SPAG9基因啟動(dòng)子區(qū)域去甲基化表達(dá)與臨床分期有關(guān),而OY-TES-1基因啟動(dòng)子區(qū)域去甲基化表達(dá)與臨床分期無(wú)關(guān);但這三種癌睪丸抗原均與上皮性卵巢癌患者的年齡、組織類(lèi)型及分化程度均無(wú)關(guān),說(shuō)明這三種癌睪丸抗原在預(yù)測(cè)上皮性卵巢癌的預(yù)后及評(píng)定上皮性卵巢癌的臨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.
【學(xué)位授予單位】：內(nèi)蒙古醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R737.31

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