基因芯片技术检测鼻咽癌紫杉醇耐药及耐药逆转相关基因的表达

目的:比较药物处理前后鼻咽癌亲本细胞系CNE-1及鼻咽癌紫杉醇耐药细胞系CNE-1/Taxol的基因表达差异,试图发现与鼻咽癌紫杉醇耐药及耐药逆转相关的基因。方法:使用基因芯片检测药物处理前后6组细胞系之间的基因表达差异,运用多重筛选和针对已知耐药相关基因的具体分析相结合的方式进行数据分析。结果:经过多重筛选,筛选出297个差异表达的基因;比较亲本细胞,在耐药细胞中上调或下调超过5倍的有17个基因。通过对已知耐药相关基因的分析,结果显示:具有药物转运作用的ATP结合盒家族中多药耐药基因(MDR1)在各组细胞中都未出现阳性表达; P450家族中CYP1A1在亲本细胞中不表达,在耐药细胞中出现较强的阳性表达,经紫杉醇处理后,其表达进一步大幅下调,顺铂处理后其表达下调;肿瘤坏死因子家族中在耐药细胞系表达下调,经顺铂处理后出现逆向表达增强的基因有TNFAIP1, 3和TNFRSF12A, 21; caspase家族出现差异表达的基因有caspase-4和caspase-6;β-微管蛋白Ⅱ在耐药细胞中表达下调;TSP1在耐药细胞中表达明显下调,经紫杉醇处理后,表达进一步下调,但是,经顺铂作用后,其表达明显上调。结论:可能与鼻咽癌紫杉醇耐药及耐药逆转相关的基因有:经过多重筛选得到的297个差异表达的基因、CYP1A1、部分肿瘤坏死因子家族成员以及另外17个在亲本细胞和耐药细胞之间表达差异超过5倍的基因;结合全部基因数据的多重筛选和已知耐药相关基因的具体分析研究肿瘤细胞耐药及耐药逆转机制是比较理想的方法。

Differential expression of taxol resistance and taxol resistance reversal related genes in nasopharyngeal carcinoma by cDNA microarray

Objective: To compare the difference in gene expression profiles between parental cell line and drug resistant cell line (CNE-1 and CNE-1/taxol) pre-treated or treated by drugs, and search for genes related to taxol resistance and reversal of taxol resistance phenotype. Methods: cDNA microarray was used to detect the difference in gene expression profiles between 6 groups of cells. Combination of multiple filtering genes and detailed analysis of documented resistance genes were used to analyze the data.Results: Through multiple filtering, 297 differentially expressed genes were screened. The expression of 17 genes was increased or decreased more than 5 folds in CNE-1/taxol compared with CNE-1.Through analyzing documented drug-resistant genes, MDR1 expression was not detected in each group. CYP1A1, one of P450 family members, was not expressed in CNE-1, but significantly increased expressions was found in CNE-1/taxol and these increased expressions were restored by cisplatin. The expression level of some members of tumor necrosis factor family was decreased in CNE-1/taxol and restored by cisplatin, including TNFAIP1, 3 and TNFRSF12A, 21. The differentially expressed members in the caspase family were caspase-4 and caspase-6. The expression of β-tubulin II was down-regulated in CNE-1/taxol. TSP1 was obviously down-regulated in CNE-1/taxol compared with CNE-1, and a more significant down-regulation of TSP1 was found when treated by taxol. However, it was greatly up-regulated after cisplatin treatment in CNE-1/taxol. Conclusion: Some genes are probably related to taxol resistance and reversal of taxol resistance in NPC cells: 297 differentially expressed genes detected by multiple filing, CYP1A1, some members of TNF family and another 17 genes whose differential expression is more than 5 folds between parental cell line and drug resistant cell line. Combination of multiple filtering genes and detailed analysis of documented resistance genes is a good method to study drug resistance and reversal of drug resistance in carcinoma cells.