克隆肺腺癌亲代细胞A549与耐顺铂细胞A549^DDP耐药相关基因的差异表达片段

目的：克隆肺腺癌耐药相关基因。方法以mRNA差异显示法技术检测耐顺铂肺腺癌细胞A549^DDP及其亲代细胞A549基因表达的差异。差异表达基因片段段被克隆并经Northern blot证实。结果获得4个基因表达的差异cDNA片段，经测序，同源性分析，其中2个片段（A1、D1）在GeneBank中未发现同源序列，一个片段（A2）与白介素－1β转化酶（Interleukin 1 β怀脲,ICE）89％同源性，一个片段（D2）与MM45s rRNA(Mouse musculus 45s pre-rRNA)基因100％同源。A1、A2cDNA片段仅表达于A549细胞，D1、D2cDNA片段仅表达于A549^DDP细胞。结论应用mRNA差异显示技术获得4个肺腺癌A549与A549^DDP间的基因差异3表达片段。2个新的差异表达片段以及与ICE、、MM45sRNA高度同源的基因片段是否与耐药相关尚需进一步研究。

Cloning of drug resistance-related gene fragments differentially expressed on lung adenocarcinoma cell line A549 and its DDP-resistant counterpart A549DDP]

OBJECTIVE: To clone drug resistance-related genes of lung adenocarcinoma. METHOD: The difference in gene expression between A549 and A549DDP was identified by mRNA differential display technique. Genes differentially expressed were cloned and confirmed by Northern blot. RESULTS: Four cDNA fragments of differentially expressed genes were obtained and sequenced. Of the 4 cDNA fragments, 2 (A1 and D1) did not show sequence homology with the existing genes in the GeneBank, one (A2) was 89% homologous with interleukin 1 beta converting enzyme (ICE) gene, and the other (D2) was identical to mouse musculus 45s pre-rRNA (MM45srRNA) gene. Both cDNA fragments A1 and A2 were expressed in A549 but not or barely expressed in A549DDP, while fragments D1 and D2 were expressed in A549DDP but not in A549. CONCLUSION: Four differentially expressed gene fragments between lung adenocarcinoma cell line and its cisplatin-resistant counterpart were identified by mRNA differential display technique. It remains to be studied whether the newly identified gene fragments and those showing sequence homology to ICE and MM45srRNA are indeed positively or negatively related to drug resistance.