摘要
目的:探讨自噬基因Beclin1在裸鼠体内对人肺腺癌A549细胞成瘤性的影响。方法:将重组质粒pRNAT-U6.2/Lenti-si423和pLenex-Beclin1通过脂质体介导的方法分别转染到A549细胞中。采用实时荧光定量PCR(real-time fluorogenic quantitative-PCR,RFQ-PCR)和蛋白质印迹法分别检测转染细胞中Beclin1mRNA和蛋白的表达;MTT法检测转染细胞在体外的增殖能力。将过表达或沉默Beclin1基因的A549细胞株分别接种于裸鼠右侧腋部皮下,观察瘤体的生长速度和大小,并采用RFQ-PCR和免疫组织化学法检测瘤体组织中Beclin1mRNA和蛋白的表达。结果:转染重组质粒pLenex-Beclin1的A549细胞中Beclin1mRNA和蛋白表达量显著增加,而体外增殖能力被明显降低;转染重组质粒pRNAT-U6.2/Lenti-si423的A549细胞中Beclin1mRNA和蛋白表达量显著减少。过表达Beclin1组的裸鼠皮下移植瘤生长缓慢,肿瘤体积明显缩小、质量明显减轻,移植瘤组织中Beclin1mRNA以及蛋白的相对表达量增加;而Beclin1基因沉默组的移植瘤组织中Beclin1mRNA及其蛋白的相对表达量降低。结论:自噬基因Beclin1的过表达可以抑制A549细胞在裸鼠体内的生长,有望成为肿瘤治疗的一个新靶点。
Objective： To investigate the effect of autophagy-related gene Beclin 1 on growth of xenografts of human lung adenocarcinoma A549 cells in BALB/c nude mice. Methods： The recombined plasmids pRNAT-U6.2/Lenti-si423 and pLenex-Beclin 1 were transiently transfected by lipofectin regeant into A549 cells, respectively. The expression levels of Beclin 1 mRNA and protein in A549 cells were detected by real-time fluorogenic quantitative-PCR （RFQ-PCR） and Western blotting, respectively. The proliferation rate of A549 cells transfected with recombinant vector pRNAT-U6.2/Lenti-si423 or pLenex- Beclin 1 was determined by MTT assay. The A549 cells expressing Beclin 1 or with Beclin 1 silencing were subcutaneously injected into right axillary region of nude mice. The growth rate and size of xenograft tumor were observed, and the expression levels of Beclin 1 mRNA and protein in xenograft tumor were determined by RFQ-PCR and immunohistochemistry, respectively. Results： The expression levels of Berlin 1 mRNA and protein were increased in A549 cells transfected with recombinant vector pLenex-Beclin 1, and the cell proliferation in vitro was inhibited. The expression levels of Beclin 1 mRNA and protein were decreased in A549 cells transfected with recombinant vector pRNAT-U6.2/Lenti-si423. As compared with Beclin 1 -silencing group, the growth rate of subcutaneous xenograft was slowing down with lower weight and smaller volume and the relative expression levels of Beclin 1 mRNA and protein were both higher in Beclin 1-overexpression group. Conclusion： Autophagy-related gene Beclin 1 can inhibit the growth of xenograft of human lung adenocarcinoma A549 cells in nude mice, and it may become a new target for tumor therapy.
出处
《肿瘤》
CAS
CSCD
北大核心
2011年第12期1061-1066,共6页
Tumor
基金
河南省科技创新人才工程资助项目(编号:0623060900)
