克隆肺腺癌亲代细胞A_(549)与耐顺铂细胞A_(549)^(DDP)耐药相关基因的差异表达片段

Cloning of drug resistancerelated gene fragments differentially expressed on lung adenocarcinoma cell line A_(549)and its DDPresistant counterpart A_(549)^(DDP)

目的 克隆肺腺癌耐药相关基因。方法 以mRNA差异显示技术检测耐顺铂肺腺癌细胞A54 9DDP及其亲代细胞A54 9基因表达的差异。差异表达基因片段被克隆并经Northernblot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 1βconvertingenzyme ,ICE) 89%同源性 ,一个片段 (D2 )与MM45srRNA(Mousemusculus 45spre rRNA)基因 10 0 %同源。A1、A2 cDNA片段仅表达于A54 9细胞 ,D1、D2 cDNA片段仅表达于A54 9DDP细胞。结论 应用mRNA差异显示技术获得 4个肺腺癌A54 9与A54 9DDP间的基因差异表达片段。 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。

Objective To clone drug resistancerelated genes of lung adenocarcinoma. Method[WT5”BZ] The difference in gene expression between A 549 and A 549 DDP was identified by mRNA differential display technique. Genes differentially expressed were cloned and confirmed by Northern blot. Results[WT5”BZ] Four cDNA fragments of differentially expressed genes were obtained and sequenced. Of the 4 cDNA fragments, 2 (A 1 and D 1) did not show sequence homology with the existing genes in the GeneBank, one (A 2) was 89% homologous with interleukin 1β converting enzyme (ICE) gene, and the other (D 2) was identical to mouse musculus 45s prerRNA (MM45srRNA) gene. Both cDNA fragments A 1 and A 2 were expressed in A 549 but not or barely expressed in A 549 DDP, while fragments D 1 and D 2 were expressed in A 549 DDP but not in A 549. Conclusion Four differentially expressed gene fragments between lung adenocarcinoma cell line and its cisplatinresistant 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.