本文選題：地西他濱 + Hela細(xì)胞　； 參考：《中華腫瘤防治雜志》2017年08期

【摘要】：目的地西他濱(decitabine,DAC)具有較好的抗腫瘤活性,其分子機(jī)制與DNA去甲基化有關(guān),本研究通過DAC對(duì)腺瘤性結(jié)腸息肉病(adenomatous polyposiscoli,APC)基因的表達(dá)水平和去甲基化的影響,探討DAC誘導(dǎo)宮頸腺癌Hela細(xì)胞凋亡分子機(jī)制。方法不同濃度的DAC作用于宮頸腺癌Hela細(xì)胞24、36和48h,MTT法檢測(cè)對(duì)宮頸癌細(xì)胞的增殖抑制作用,觀察DAC對(duì)Hela細(xì)胞增殖的濃度依賴效應(yīng)和時(shí)間依賴效應(yīng)。流式細(xì)胞術(shù)檢測(cè)DAC對(duì)宮頸腺癌Hela細(xì)胞凋亡和細(xì)胞周期的影響。在DAC作用于宮頸腺癌Hela細(xì)胞前后,通過甲基化特異性PCR(Methylation specific PCR,MSP)檢測(cè)APC基因的甲基化狀態(tài);RT-PCR法檢測(cè)APC基因mRNA表達(dá)水平;蛋白質(zhì)印跡法檢測(cè)APC蛋白、β-catenin蛋白在胞內(nèi)及核內(nèi)表達(dá)的變化。結(jié)果 DAC對(duì)宮頸腺癌Hela細(xì)胞增殖抑制具有濃度依賴效應(yīng)和時(shí)間依賴效應(yīng),Hela細(xì)胞半數(shù)抑制濃度(IC50)24、36、48h分別為28.23、7.65和5.64μmol/L。DAC處理后的宮頸腺癌Hela細(xì)胞的凋亡率為(73.82±0.11)%,明顯高于對(duì)照組的(12.41±0.24)%,P0.001。DAC處理Hela細(xì)胞后,S期細(xì)胞比例(47.82±2.57)%和G2期細(xì)胞比例(30.87±2.28)%顯著高于空白對(duì)照組的S期細(xì)胞比例(24.08±0.71)%和G2期細(xì)胞比例(2.52±0.84)%,P0.001。DAC處理后,APC基因啟動(dòng)子區(qū)域去甲基化狀態(tài)明顯增高,APC mRNA表達(dá)量上升,處理前后比較差異有統(tǒng)計(jì)學(xué)意義,P0.05。DAC處理后,胞內(nèi)APC蛋白表達(dá)上調(diào),而胞內(nèi)和核內(nèi)的β-catenin蛋白表達(dá)下調(diào),差異均有統(tǒng)計(jì)學(xué)意義,P0.05。結(jié)論 DAC可通過對(duì)APC基因的去甲基化作用,上調(diào)胞內(nèi)APC蛋白表達(dá),下調(diào)胞內(nèi)和核內(nèi)β-catenin表達(dá),誘導(dǎo)宮頸癌細(xì)胞凋亡。
[Abstract]:Decitabine DAC has good antitumor activity and its molecular mechanism is related to DNA demethylation. In this study, the effect of DAC on the expression and demethylation of adenomatous polyposis gene was studied. To investigate the molecular mechanism of apoptosis induced by DAC in Hela cells of cervical adenocarcinoma. Methods the inhibitory effects of DAC on the proliferation of cervical carcinoma Hela cells were detected by MTT assay at 24 ~ 36 and 48 h, and the concentration-dependent and time-dependent effects of DAC on Hela cell proliferation were observed. The effect of DAC on apoptosis and cell cycle of Hela cells in cervical adenocarcinoma was detected by flow cytometry. The expression level of APC mRNA was detected by methylation specific PCR, and the expression of APC protein and 尾 -catenin protein in cell and nucleus were detected by Western blotting. Results DAC had concentration-dependent and time-dependent effects on the proliferation inhibition of Hela cells. The apoptotic rate of Hela cells treated with DAC was 28.237.65 and 5.64 渭 mol / L / L DAC for 48 h, respectively, which was significantly higher than that in the control group (73.82 鹵0.11). The percentage of S phase cells and G2 phase cells in Hela cells treated with P0.001.DAC were significantly higher than those in blank control group (24.08 鹵0.71)% and (2.52 鹵0.84) P0.001.DAC, respectively. The percentage of S phase cells and G 2 phase cells in Hela cells treated with P0.001.DAC was significantly higher than that in control group (30.87 鹵2.28)%, and the percentage of APC gene promoter region in Hela cells treated with DAC was significantly higher than that in blank control group (24.08 鹵0.71)% and (2.52 鹵0.84) P0.001.DAC treatment. The expression of APC mRNA increased, After treatment with P0.05.DAC, the expression of APC protein was up-regulated, while the expression of 尾 -catenin was down-regulated in the nucleus and intracellular cells, and the difference was statistically significant (P 0.05). Conclusion DAC can up-regulate the expression of APC protein, down-regulate the expression of 尾 -catenin and induce apoptosis of cervical cancer cells by demethylation of APC gene.
【作者單位】： 南華大學(xué)附屬南華醫(yī)院婦產(chǎn)科;
【分類號(hào)】：R737.33

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