Yu Feng, Li Hua, Bu Xuefeng, Zhang Yongjun
Department of General Surgery, Jiangsu University, Jiangsu Province, China.
Hepatogastroenterology. 2012 Mar-Apr;59(114):561-4. doi: 10.5754/hge10760.
BACKGROUND/AIMS: To investigate the effects of integrin-β1 gene expression inhibited by shRNA on invasion of pancreatic carcinoma PANC-1 cells in vitro.
The eukaryotic expression plasmid of short hairpin RNA (shRNA) targeting integrin-β1 gene (integrin-β1-shRNA) was constructed and transfected into PANC-1 cells. The expressions of integrin-β1 mRNA and protein were detected by real-time quantitative polymerase chain reaction (PCR) and western blot assay, respectively. The invasive ability of PANC-1 cells was observed with a transwell cell culture chamber and the expressions of MMP-2 and MMP-9 were assayed.
Compared to the untransfected group, recombinant expression plasmid integrin-β1-shRNA resulted in reduction of integrin-β1 mRNA and protein by 78.58%±7.24% and 92.88%±3.18%, respectively and the average number of invading PANC-1 cells were decreased from 52±5 to 21±4 (p<0.01) and the expressions of MMP-2 and MMP-9 were inhibited.
Recombinant expression plasmid integrin-β1- shRNA can inhibit the expression of integrin-β1 gene and suppress the invasion of PANC-1 cells in vitro significantly.
背景/目的：研究shRNA抑制整合素β1基因表达对胰腺癌PANC-1细胞体外侵袭能力的影响。
构建靶向整合素β1基因的短发夹RNA（shRNA）真核表达质粒（整合素β1-shRNA），并转染至PANC-1细胞。分别采用实时定量聚合酶链反应（PCR）和蛋白质印迹法检测整合素β1 mRNA和蛋白的表达。使用Transwell细胞培养小室观察PANC-1细胞的侵袭能力，并检测基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）的表达。
与未转染组相比，重组表达质粒整合素β1-shRNA使整合素β1 mRNA和蛋白分别降低了78.58%±7.24%和92.88%±3.18%，侵袭的PANC-1细胞平均数从52±5降至21±4（p<0.01），且MMP-2和MMP-9的表达受到抑制。
重组表达质粒整合素β1-shRNA能够抑制整合素β1基因的表达，并显著抑制PANC-1细胞的体外侵袭。
