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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 resistance-related genes in lung adenocarcinoma. Methods mRNA differential display technique was used to detect the difference of gene expression of cisplatin-resistant lung adenocarcinoma cell line A54 9DDP and its parental cell A549. Differentially expressed gene fragments were cloned and confirmed by Northern blot. Results Four differentially expressed cDNA fragments were obtained. After sequencing and homology analysis, two fragments (A1, D1) had no homologous sequences in GeneBank. One fragment (A2) and interleukin 1β converting enzyme (ICE) 89% homology. One fragment (D2) is 100% homologous to the MM45 sRNA (Mousemusculus 45s pre rRNA) gene. The A1 and A2 cDNA fragments were only expressed in A54 9 cells, and the D1 and D2 cDNA fragments were only expressed in A54 9DDP cells. Conclusion The mRNA differential display technique was used to obtain differentially expressed genes between four lung adenocarcinomas A54 9 and A54 9DDP. Two new differentially expressed fragments and whether the gene fragments highly homologous to ICE and MM45sRNA are related to drug resistance still need further study.