摘要
目的 探讨肺腺癌SPCA-1细胞中多药耐药相关蛋白(MRP)基因甲基化状态与MRPmRNA、MRP表达的关系。方法 应用限制性内切酶Eco47Ⅲ和PCR法检测人胚肺WI-38细胞、肺腺癌SPCA-1细胞及经阿霉素诱导的耐药细胞MRP基因甲基化状态;原位杂交法检测MRPmRNA表达;免疫组化法测定MRP表达。结果 WI-38细胞MRP基因启动子区处于高甲基化状态,SPCA-1肺腺癌细胞和经阿霉素诱导的耐药细胞MRP基因启动子区处于低甲基化状态。WI-38细胞、SPCA-1肺腺癌细胞、不同浓度阿霉素诱导的耐药SPCA-1细胞MRPmRNA和MRP表达的阳性率和强阳性率,经组间多重比较均有显著性差异。结论 SPCA-1肺腺癌细胞MRP基因启动子区处于低甲基化状态,尤其5’端调控区低甲基化是导致转录活性增加的重要结构基础,参与肺癌耐药的发生。
Objective To investigate the relation between the methylation status of multi - drug resistance protein gene ( MRP gene) and the expression of its mRN A and multi - drug resistance protein ( MRP) in the cell line SPCA -Ⅰ of lung adenocacinoma. Methods Cell line WI - 38 , cell line SPG A -1 and its drug - resistant cells induced by different concentration of doxorubicin were treated with restriction endonuclease Eco47Ⅲ, the methylation status of the MRP gene was examined by PCR, and the expression of mRNA and MRP were detected by in situ hybridization and immunohistochemistry. Results The status of the MRP gene in cell line WI - 38 at the promoter region was hypermethylated, but the status of MRP gene in lung adenocacinoma cell line SPCA - Ⅰ and it' s drug - resistant cells induced by different concentrations doxorubicin at the promoter region were hypomethylated. There was significant difference in the expression of the positive ratio and strong positive ratio of the expression of mRNA in cell line WI - 38, cell line SPCA -Ⅰand its drug - resistant cells induced by different concentration of doxorubicin (P =0. 000). Furthermore, it was also significant difference in the expression positive ratio and strong positive ratio pres-sion in cell line WI - 38, SPCA -Ⅰ and its drug - resistant cells induced by different concentration of MRP exdoxorubicin(P=0.000, P=0.002). Conclusion The status of MRP gene in cell line SPCA-Ⅰ of lung adenocacinoma at the promoter region was hypomethylation. Especially, the hypomethy-lation of its 5 ' regulaion region at the promoter is an important basic structure that increase the activity of transcriptand is related with the development of drug resistance in lung cancer.
出处
《上海第二医科大学学报》
CSCD
2003年第B10期5-8,共4页
Acta Universitatis Medicinalis Secondae Shanghai
基金
上海市高等学校科学技术发展基金(99B18)
关键词
肺腺癌
MRP基因
甲基化
多药耐药
相关性
癌细胞
lung neoplasms
multi - drug resistance protein gene
methylation
multi - drug resistance
