CAJ | 학술논문

目的:通过 RNA 干扰技术沉默 B 淋巴细胞瘤-2 基因 ( Bcl2 ) 和切除修复交叉互补基因 1 ( Ercc1 )并观察对肺癌细胞株增殖的影响. 方法:分别设计特异性沉默 Bcl2 和 Ercc1 的短发夹 RNA 序列,并克隆到腺病毒骨架载体上形成重组腺病毒质粒. 将质粒转染到 HEK293 细胞中,包装和扩增形成完整高滴度的重组腺病毒, 将腺病毒感染人肺癌 H460 细胞株, 流式细胞术检测抑制 Ercc1 和 Bcl2 基因后肺癌细胞凋亡变化. 结果:通过酶切鉴定,证实重组腺病毒载体构建成功, PCR 鉴定扩增出 1 200 bp 的目的条带,病毒滴度为 5 × 1011/mL. 流式细胞术检测结果显示,AdsiBcl2-Ercc1 组细胞凋亡率比单基因干扰组增高.结论:成功构建干扰 Bcl2 和 Ercc1 的重组质粒及重组腺病毒,同时沉默 Bcl2 和 Ercc1 可以促进肺癌细胞凋亡.
목적:통과 RNA 간우기술침묵 B 림파세포류-2 기인 ( Bcl2 ) 화절제수복교차호보기인 1 ( Ercc1 )병관찰대폐암세포주증식적영향. 방법:분별설계특이성침묵 Bcl2 화 Ercc1 적단발협 RNA 서렬,병극륭도선병독골가재체상형성중조선병독질립. 장질립전염도 HEK293 세포중,포장화확증형성완정고적도적중조선병독, 장선병독감염인폐암 H460 세포주, 류식세포술검측억제 Ercc1 화 Bcl2 기인후폐암세포조망변화. 결과:통과매절감정,증실중조선병독재체구건성공, PCR 감정확증출 1 200 bp 적목적조대,병독적도위 5 × 1011/mL. 류식세포술검측결과현시,AdsiBcl2-Ercc1 조세포조망솔비단기인간우조증고.결론:성공구건간우 Bcl2 화 Ercc1 적중조질립급중조선병독,동시침묵 Bcl2 화 Ercc1 가이촉진폐암세포조망.
Objective Toexplore the effect of Bcl2 and Ercc1 gene on apoptosis by RNA interference of Bcl2 and Ercc1 gene expression in non-small cell lung cancer cell line. Methods shRNA targeting Bcl2 and Ercc1 gene were cloned into vector to construct recombinant adenovirus plasmids which were transfected into HEK293 cells. High titer adenovirus was obtained after package and amplification. H460 cells were infected with the adenovirus and apoptosis rate was detected by flow cytometry. Results The results of restriction endonuclease digestion showed that recombinantadenovirus vector adsiBcl2-Ercc1 was successfully constructed. The positive 1 200 bp amplification bands could be seen in PCR analysis , and the titer of the adenovirus was 5 × 1011/mL. The apoptosis rate of AdsiBcl2-Ercc1 was higher than that of the two single silenced gene groups. Conclusions The recombinant plasmids and recombinant adenovirus were successfully constructed. Silencing of Bcl2 and Ercc1 through shRNA promotes the apoptosisin lung cancer cells.