医学高级职称论文.ppt

5-Aza-CdR 对人食管癌细胞株生物学行为及 TFPI-2 mRNA 表达的影响 5-Aza-CdR 对人食管癌细胞株生物学行为及 TFPI-2 mRNA 表达的影响作者:杜雅冰,邵应举,樊青霞,孙桢,王琳,赵培荣作者单位: (郑州大学第一附属医院肿瘤内科,河南郑州 450002) 【摘要】目的探讨 5-氮杂-2- 脱氧胞苷(5-Aza-2-deoxycytidine,5-Aza- CdR) 干预对食管鳞癌细胞株 Eca9706 生长增殖及其组织因子途径抑制物 2(tissue factor pathway inhibitor-2 , TFPI-2) 基因表达的影响。方法选取食管鳞癌细胞株 Eca9706 ,并用不同浓度的 5-Aza-CdR 处理该细胞株。 MTT 法检测干预前后细胞增长率的变化, FCM 法检测干预前后细胞凋亡率的变化, RT-PCR 技术检测干预前后 Eca9706 细胞 TFPI-2 基因 mRNA 的表达,免疫组化检测干预前后 Eca9706 细胞 TFPI-2 蛋白的表达。结果 食管鳞癌细胞株 Eca9706 存在 TFPI-2 基因超甲基化状态,经 5-Aza-CdR 处理后,超甲基化状态解除,细胞增殖受到抑制,细胞凋亡率明显提高(P<); 细胞中 TFPI-2 蛋白表达明显增强,同时 mRNA 的表达水平也较处理前明显提高(P<) 。结论食管癌细胞系 TFPI-2 基因超甲基化可抑制其 mRNA 表达,当其超甲基化解除后,细胞增殖受到抑制,同时细胞凋亡率相应提高。【关键词】食管癌;组织因子途径抑制物-2;5- 氮杂-2- 脱氧胞苷;甲基化;免疫组化;逆转录聚合酶链反应;凋亡 ABSTRACT : Objective To explore the effects of 5-Aza-2-deoxycytidine (5-Aza-CdR) intervention on the growth and proliferation of Eca9706 cell line and protein expression of tissue factor pathway inhibitor-2 (TFPI-2). Methods Eca9706 cell line was treated by 5-azaCdR of different concentrations; MTT , flow cytometry , immunohistochemistry and RT-PCR were used to determine the cell growth , apoptosis , and the expression of TFPI-2 gene and its protein. Results Eca9706 cell line had hypermethylation of TFPI-2. After demethylation by 5-Aza-CdR treatment , the proliferation of Eca9706 was inhibited , the apoptosis rate was also increased significantly in the concentration-dependent manner. RT-PCR detected that mRNA expression of TFPI-2 gene in Eca9706 cell line recovered significantly (P<). Conclusion 5-Aza-CdR can slow the growth of Eca9706 cell , increase the apoptosis rate , reactivate the TFPI-2 gene transcription and protein expression by demethylation. KEY WORDS : esophageal carcinoma; tissue factor pathway inhibitor-2; 5-Aza-CdR; methylation; immunohistochemistry; RT-PCR; apoptosis 组织因子途径移植物 2(tissue factor pathway inhibitor , TFPI-2) 是丝氨酸蛋白酶抑制物,在调控肿瘤细胞浸润转移方面起着重要作用。目前,关于 TFPI-2 在食管癌中的基因表达情况的分析研究尚少。本研究旨在探讨 TFPI-2 基因的失活机制,