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目的:探讨人胰腺癌细胞株HHIP表达与其甲基化的关系,为胰腺癌发生机制的研究提供新的信息。方法:以人胰腺癌细胞株BxPC-3、CFPAC-1、PANC-1、AsPC-1和PaTu8988s为研究对象,用反转录聚合酶链反应(RT-PCR)及免疫细胞化学染色(S-P)法检测去甲基化制剂——DNA甲基转移酶抑制剂5-氮杂-2′-脱氧胞苷(5-Aza-CdR)处理前后各胰腺癌细胞株HHIPmRNA/蛋白表达的变化,用甲基化特异性PCR(MSP)结合测序检测HHIP基因CpG岛甲基化状态。结果:5-Aza-CdR处理前,胰腺癌细胞株BxPC-3、CFPAC-1、PANC-1、AsPC-1和PaTu8988s无HHIPmRNA/蛋白表达;经5-Aza-CdR处理后,HHIPmRNA/蛋白重新表达。MSP结合测序显示上述胰腺癌细胞株HHIP基因CpG岛存在高甲基化。结论:人胰腺癌细胞株HHIP表达抑制与其基因CpG岛高甲基化相关。HHIP基因CpG岛高甲基化可能在胰腺癌的发生、发展中起一定作用。
Objective: To investigate the relationship between HHIP expression and methylation in human pancreatic cancer cell lines, and to provide new information for the study on the mechanism of pancreatic cancer. Methods: The human pancreatic cancer cell lines BxPC-3, CFPAC-1, PANC-1, AsPC-1 and PaTu8988s were used as research objects. Reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry (SP) Method was used to detect the changes of HHIPmRNA / protein expression in pancreatic cancer cell lines before and after demethylation agent-5-Aza-2’-deoxycytidine (5-Aza-CdR) The methylation status of CpG island of HHIP gene was detected by MSP and sequencing. Results: HHIP mRNA and protein were not expressed in pancreatic cancer cell lines BxPC-3, CFPAC-1, PANC-1, AsPC-1 and PaTu8988s before treatment with 5-Aza-CdR. expression. MSP binding showed that there was hypermethylation in the CpG island of HHIP gene in the above pancreatic cancer cell lines. Conclusion: Inhibition of HHIP expression in human pancreatic cancer cell line is associated with hypermethylation of CpG island. HHIP gene CpG island hypermethylation may play a role in the occurrence and development of pancreatic cancer.