摘要
目的研究缺氧诱导因子-1α（HIF-1α）对低氧状态下前列腺癌PC-3细胞株增殖及侵袭的影响。方法利用转染试剂Fermentas将人HIF-1α重组表达质粒pcDNA3．1—HIF--1α转染PC-3细胞株后，低氧环境培养，采用G418筛选，建立稳定表达HIF-1α基因的细胞株，分别命名为pcD—NA3.1-HIF-1α-PC-3、pcDNA3．1-PC-3及PC-3组。采用RT—PCR和蛋白质印迹法检测3组细胞HIF-1α mRNA和蛋白的表达情况；噻唑盐法测定细胞生长；transwell小室检测侵袭能力。结果与pcDNA3．1-PC-3组和PC-3组相比，pcDNA3．1-HIF--1α-PC-3组细胞内HIF-1αmRNA条带增强不明显，pcDNA3．1-HIF-1α—PC-3组细胞内HIF-1α蛋白的条带明显增强，HIF-1α过表达的PC-3细胞增殖速度明显增快，侵袭细胞数明显增多。结论HIF--1α过表达对PC-3细胞株的增殖及侵袭具有促进作用。
Objective WE transfected the recombinant expression plasmid of pcDNA3. 1-HIF-1α into the prostate cancer cells, to research the effect of HIF-1α on proliferation of prostate cancer cell PC-3. Methods We selected a stable expression cell line with G418 were selected by transfection of the recombi- nant expression plasmid of pcDNA3.1-HIF-1α into the prostate cancer. The protein and mRNA expression of HIF-1α was assayed by western -blot and RT-PCR. The cells growth curves were described by MTT and the ability of invasion was assayed by Transwell. Results The expression of HIF-1α mRNA was not obviously increased compared to the untransfected prostate cancer cell by RT-PCR, but the expression of HIF-1 ct protein was up-regulated by western-blot after the recombinant expression plasmid transfected into PC-3. The ability of cell proliferation and invasion was significantly enhanced by MTT and Transwell assays. Conclusion The stable expression cell model of HIF-1α was successfully constructed, which enhanced the proliferation and invasion of prostate cancer cell PC-3.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2011年第9期599-602,共4页
Chinese Journal of Urology
基金
厦门市科技局科研基金资助（3502220084002）
