摘要
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 explore the relationship between HHIP expression and its hypermethylation in human pancreatic cancer cell lines,so as to provide new insights into the tumorigenesis of pancreatic cancer.Methods： Human pancreatic cancer cell lines BxPC-3,CFPAC-1,PANC-1,AsPC-1 and PaTu8988s were used in the present study. Reverse transcriptase polymerase chain reaction（RT-PCR） and immunocytochemical S-P method were used to detect the expression of HHIP at mRNA and protein levels before and after treatment with 5-aza-2′-deoxycytidine （5-Aza-CdR）,a DNA methyltransferase inhibitor.Methylation-specific PCR （MSP） combined with DNA sequencing were adopted to identify the CpG island methylation of HHIP gene.Results： Expression of HHIP mRNA/protein was absent in the five pancreatic cancer cell lines before 5-Aza-CdR treatment, and re-expressed after treatment with 5-Aza-CdR.CpG island hypermethylation of HHIP gene was observed in the five pancreatic cell lines by MSP and DNA sequencing.Conclusion： Hypermethylation in CpG island of HHIP gene is correlated with the inhibition of HHIP expression in human pancreatic cancer cell lines, and it might play an important role in the development and progression of pancreatic cancer.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2009年第8期879-883,共5页
Academic Journal of Second Military Medical University
基金
上海市卫生局青年科研项目基金(2007Y85)~~
