本文選題：膀胱癌 + 甲基化　； 參考：《安徽醫(yī)科大學(xué)學(xué)報(bào)》2017年07期

【摘要】：目的檢測(cè)生長(zhǎng)分化因子15(GDF15)在膀胱癌中的甲基化狀態(tài),探討其啟動(dòng)子區(qū)異常甲基化對(duì)于膀胱癌發(fā)生發(fā)展的作用。方法應(yīng)用重亞硫酸鹽測(cè)序PCR(BSP)聯(lián)合T-載體PCR產(chǎn)物(TA)克隆檢測(cè)人膀胱移行細(xì)胞癌細(xì)胞系5637、HT1376、KU19-19、膀胱癌組織、癌旁組織、正常組織樣品中GDF15啟動(dòng)子區(qū)甲基化狀態(tài),并用甲基化酶抑制劑5-Aza-2-deoxycitydine(5-Aza-dc)處理5637,觀察處理前后平均甲基化率的變化情況,Western blot法檢測(cè)5637處理前后蛋白表達(dá)情況,MTT法檢測(cè)細(xì)胞增殖情況,劃痕實(shí)驗(yàn)檢測(cè)遷移情況,Transwell法檢測(cè)侵襲能力。結(jié)果 5637、HT1376、KU19-19細(xì)胞中GDF15啟動(dòng)子區(qū)平均甲基化率為89.29%、10.71%、8.33%,腫瘤組織、癌旁組織及正常組織分別為86.91%、9.52%、5.95%,膀胱癌組織中GDF15啟動(dòng)子區(qū)甲基化率較癌旁組織和正常組織中高,差異有統(tǒng)計(jì)學(xué)意義(P0.05);5-Aza-dc可以逆轉(zhuǎn)5637中GDF15的甲基化狀態(tài):與處理前相比,處理組5637細(xì)胞系GDF15蛋白表達(dá)增加,增殖、遷移、侵襲能力下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論膀胱癌中GDF15基因表達(dá)與甲基化狀態(tài)有關(guān),啟動(dòng)子區(qū)高甲基化導(dǎo)致基因沉默,逆轉(zhuǎn)甲基化狀態(tài)可以使基因蛋白表達(dá)增加,細(xì)胞增殖、遷移、侵襲能力均降低。GDF15基因甲基化異常狀態(tài)有可能成為膀胱癌診斷的潛在靶點(diǎn)。
[Abstract]:Objective to detect the methylation status of growth differentiation factor 15 (GDF 15) in bladder cancer and to explore the role of abnormal methylation in its promoter region in the occurrence and development of bladder cancer. Methods the methylation status of GDF15 promoter was detected in human bladder transitional cell carcinoma cell line 5637 (HT1376) KU19-19 by PCRS-BSPand T- vector PCR product (TAA). The methylation status of GDF15 promoter was detected in bladder cancer tissues, adjacent tissues and normal tissues. 5637 was treated with 5-Aza-2-deoxycineine (5-Aza-2-deoxycineine). The changes of average methylation rate before and after treatment were observed. Western blot assay was used to detect protein expression before and after treatment. MTT assay was used to detect cell proliferation, and scratch test was used to detect migration and invasion ability. Results the average methylation rate of GDF15 promoter in 5637 HT1376 KU19-19 cells was 89.290.71 and 8.33. The methylation rate of GDF15 promoter in bladder cancer was higher than that in paracancerous and normal tissues. The difference was statistically significant (P 0.05) that 5-Aza-dc could reverse the methylation of GDF15 in 5637. Compared with before treatment, the expression of GDF15 protein increased, proliferation, migration and invasion decreased in 5637 cell line of treatment group, and the difference was statistically significant (P 0.05). Conclusion GDF15 gene expression is related to methylation status in bladder cancer. Hypermethylation in promoter region leads to gene silencing. Reverse methylation can increase gene protein expression, cell proliferation and migration. Decreased invasiveness of the. GDF15 gene methylation may be a potential target for the diagnosis of bladder cancer.
【作者單位】： 安徽醫(yī)科大學(xué)附屬安徽省立醫(yī)院泌尿外科;
【基金】：安徽省自然科學(xué)基金(編號(hào):1608085MH166)
【分類號(hào)】：R737.14

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本文編號(hào)：1810480