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醫(yī)學(xué)免費(fèi)論文:天花粉蛋白誘導(dǎo)宮頸癌HeLa細(xì)胞凋亡及APC基因去甲基化
【摘要】 目的:研究天花粉蛋白(TCS)抗宮頸癌作用機(jī)制。方法:MTT還原法檢測(cè)TCS對(duì)宮頸癌HeLa細(xì)胞的抑制作用，流式細(xì)胞儀檢測(cè)TCS對(duì)HeLa細(xì)胞周期及凋亡率的影響，MSP檢測(cè)TCS作用前后APC基因啟動(dòng)子甲基化狀態(tài)。結(jié)果:TCS對(duì)HeLa細(xì)胞的生長(zhǎng)具有明顯的抑制作用，呈劑量依賴性;TCS可以將HeLa細(xì)胞阻滯于S期;TCS誘導(dǎo)HeLa細(xì)胞發(fā)生凋亡,有劑量依賴性;80 mg/L TCS處理HeLa細(xì)胞48 h后， APC基因啟動(dòng)子出現(xiàn)去甲基化。結(jié)論:TCS對(duì)HeLa細(xì)胞有明顯抑制作用，抑制機(jī)理與將HeLa細(xì)胞阻滯于S期、誘導(dǎo)細(xì)胞凋亡及APC基因啟動(dòng)子出現(xiàn)去甲基化有關(guān)，TCS有良好的抗宮頸癌臨床應(yīng)用前景。
【關(guān)鍵詞】 天花粉蛋白;HeLa細(xì)胞;細(xì)胞凋亡;APC基因
Effects of Trichosanthin on Inducing Apoptosis of Hela Cells and Methylation of APC Gene in Cervical CancerCHEN Yong,ZHENG Rong,LIU Yong-zhen,CHEN Dong-li,CHEN Shuang-yun^ (Department of Traditional Chinese Medicine,Hubei College of Traditional Chinese Medicine,Wuhan,Hubei 430061;Department of Gynecology,Taihe Hospital,Yunyang Medical College,Shiyan,Hubei 442000,China)Abstract:Objective To investigate the mechanism of trichosanthin(TCS) against cervical carcinoma.Methods The inhibitory effect of TCS on Hela cells proliferation was detected by MTT assay.The cell cycle and apoptosis rate of Hela cells were determined by flow cytometry.The methylation status of APC gene was determined with MSP before and after treatment respectively.Results The growth of Hela cells was significantly inhibited with TCS and the inhibitory effect was dose-dependent;the differentiation of Hela cells was blocked at S-phase of the cell cycle;the apoptosis of Hela cells was induced with TCS in a dose-dependent manner;The demethylation of APC gene promoter was observed after treating with TCS at 80 mg/L for 48 h.Conclusion Trichosanthin could inhibit the proliferation of Hela cells,which may be related with the cell cycle block,apoptosis induction and demethylation of APC gene promoter.TCS may have a wide clinical respect in cervical cancer treatment醫(yī).學(xué)全.在.線網(wǎng)站zxtf.net.cn.
Key words:Trichosanthin;HeLa cells;Apoptosis;APC gene
天花粉蛋白(Trichosanthin，TCS)為中藥天花粉的有效成分, 具有多種藥理學(xué)活性，已應(yīng)用于臨床。近年來(lái)通過(guò)對(duì)天花粉蛋白結(jié)構(gòu)和功能的深入研究,發(fā)現(xiàn)其具有抗腫瘤的作用。 TCS可用于臨床抗絨癌及葡萄胎的治療,并對(duì)多種腫瘤細(xì)胞(如胃癌、肝癌、結(jié)直腸癌、黑素瘤和白血病細(xì)胞等)有顯著的殺傷作用,是一種有前途的抗腫瘤藥物和免疫毒素彈頭[1]。本研究應(yīng)用TCS作用于宮頸癌HeLa細(xì)胞系，觀察其對(duì)細(xì)胞增殖及凋亡的影響，并研究其對(duì)HeLa細(xì)胞腺瘤性結(jié)腸息肉病(adenomatous polyposis coli,APC)基因甲基化狀態(tài)的影響，探討TCS抗宮頸癌作用及其機(jī)制。
1 材料和方法
1.1 藥品和試劑
天花粉蛋白注射液(1.2 g/L)(上海金山制藥有限公司)，新生牛血清(四季青)，培養(yǎng)基DMEM(GBICO公司)，細(xì)胞毒性檢測(cè)試劑盒及PI染料試劑盒(凱基生物科技發(fā)展有限公司)，組織基因組DNA提取試劑盒及DNA Ladder(北京天根生化科技有限公司)，DNA甲基化試劑盒(北京天漠科技開(kāi)發(fā)有限公司)，Taq DNA聚合酶(Promega公司)，dNTP Mix(Amresco公司)，引物合成(上海生工生物工程技術(shù)服務(wù)有限公司)。
