BRI基因在一对转移能力不同的肺腺癌细胞系中的序列分析与表达研究

Sequence analysis and expression study of BRI gene in two lung adenocarcinoma cell lines with different metastatic potential

目的 确认淀粉样纤维蛋白基因 (amyloid fibrils,BRI)基因在 1对同源但转移能力不同的肺腺癌细胞系 AGZY83- a和 Anip973中的序列并分析其表达。方法 采用测序技术 ,Northern印迹杂交 ,G显带后荧光原位杂交分析 BRI基因在肺腺癌细胞系的序列与表达。结果 BRI基因在高转移肺腺癌细胞系 Anip973中高表达 ,在其低转移母系 AGZY83- a中低表达 ,两细胞系 BRI基因染色体定位区均存在断裂重排 ,该基因染色体定位区在 Anip973中出现扩增。已知 BRI基因的 - 116 bp～ - 5 bp处碱基序列和- 115 bp～ - 5 bp处碱基序列在 AGZY83- a和 Anip973中分别突变为 CTCAGCAGCCCGC和 TCAGC-CGC。结论 BRI基因在转移能力不同的肺腺癌细胞系差异表达与该基因的染色体定位区域的断裂重排无关 ,与该基因染色体定位区拷贝数增加及 5′非翻译区存在不同的突变可能相关。

Objective: To investigate the sequence of amyloid fibrils (BRI) gene and its expression in two lung adenocarcinoma cell lines AGZY83-a and Anip973 with the same tumor origin but different metastatic potential. Methods: DNA sequencing, sequential G banding fluorescence in situ hybridization (FISH) and Northern blot were used to analyze the sequence and expression of BRI gene was in two lung adenocarcinoma cell lines with different metastatic potential. Results: The expression of BRI gene was up-regulated in the highly metastatic cell line Anip973 and was down-regulated in the low metastatic cell line AGZY83-a from which the Anip973 was derived. FISH results disclosed that in the two cell lines, the same rearrangements existed in the chromosome region where BRI gene was located, but in Anip973, amplification took place in the chromosome region where BRI gene was located. DNA sequencing results showed different mutations in the 5′ untranslated region of BRI gene in the two cell lines. Conclusion: The above results revealed that there was no relation between BRI gene differential expression and rearrangements of chromosome. The amplification of the chromosome region where BRI gene was located and the different mutations in the 5′ untranslated region of BRI gene probably contributed to the differential expression.