卵巢癌卡铂耐药相关肿瘤抑制基因差异表达的研究

Differential Expression of Tumor Suppressor Gene in Carboplatin Resistance Microarray of Ovarian Carcinoma

目的分析卵巢癌卡铂耐药基因芯片中卡铂耐药细胞系SKOV3-CB较其亲本SKOV3细胞系差异表达下调的肿瘤抑制基因,并验证其差异表达。方法应用分子生物信息学方法 ,分析基因芯片中卵巢癌SKOV3-CB细胞系较SKOV3细胞系差异表达下调两倍以上的基因1554条,筛选出肿瘤抑制基因17个,即:RNASET2,VHL,DLG1,COPS2,NOL7,GGNBP2,RBL1,S100A2,PARG1,PERP,TCF3,DNAJA3,PCGF2,ING1,CYLD,RARRES3,RBBP8,应用荧光定量PCR技术对差异表达进行验证。结果 RNASET2,VHL,DLG1,COPS2,NOL7,RBL1,S100A2,PARG1,PERP,TCF3,DNAJA3,ING1,CYLD,RARRES3,RBBP8基因差异表达下调,下调倍数分别为24.47、65.24、10.81、21.30、72.94、5.95、27.13、1.57、1.32、12.93、46.80、7.57、14.90、14.00、23.98。GGNBP2,PCGF2,DLG1基因差异表达上调,上调倍数分别为5.76、2.19和17.10,基因芯片差异表达结果与real-timePCR所验证表达下降趋势的结果符合率为82.35%,下调两倍以上的基因符合率为64.70%。结论利用基因芯片技术分析基因差异表达,结合real-timePCR对其检测结果验证是一种有效的方法 ,多肿瘤抑制基因参与卵巢癌卡铂耐药的机制。

Objective To analyze the down-regulated tumor suppressor gene of differential expressions of carboplatin resistance cell line SKOV3-CB and its parental cell line SKOV3 in ovarian cancer carboplatin resistance gene array.Methods Using molecular bioinformatics methods to analyze 1554 over-2-fold-downregulated genes of the differential expressions of ovarian cancer SKOV3-CB cell lines and SKOV3 cell lines,and 17 tumor suppressor genes （RNASET2,VHL,DLG1,COPS2,NOL7,GGNBP2,RBL1,S100A2,PARG1,PERP,TCF3,DNAJA3,PCGF2,ING1,CYLD,RARRES3,RBBP8） were screened out,and the differential expressions were verified by real-time quantitative PCR technique.Results The differential expressions of genes RNASET2,VHL,DLG1,COPS2,NOL7,RBL1,S100A2,PARG1,PERP,TCF3,DNAJA3,ING1,CYLD,RARRES3,RBBP8 were down regulated,the multiples of which were 24.47,65.24,10.81,21.30,72.94,5.95,27.13,1.57,1.32,12.93,46.80,7.57,14.90,14.00,23.98,respectively.The differential expressions of genes GGNBP2,PCGF2,DLG1 were up regulated,the multiples of which were 5.76,2.19,17.10,respectively.The coincidence of the results of gene microarray differential expressions with those of downward trend of expressions validated by real-time PCR was 82.35%,that of genes down-regulated 2 folds was 64.70%.Conclusion The gene differential expression analysis by cDNA microarray technique combined with test result verification by real-time PCR is an effective method.Multiple tumor suppressor genes were involved in carboplatin resistance mechanism of ovarian cancer.