Inhibitory effects of adenovirus-mediated shRNA targeting Akt1 and COX-2 on invasion and metastasis of SGC-7901 cells
10.3760/cma.j.issn.1007-631X.2009.09.017
- VernacularTitle:腺病毒介导的靶向蛋白激酶B1和环氧合酶2短发夹RNA抑制人胃腺癌细胞的侵袭和转移
- Author:
Jing ZHANG
;
Yanchao FU
;
Chunsheng KANG
;
Qingyu ZHANG
;
Tao WANG
;
Jie ZHANG
- Publication Type:Journal Article
- Keywords:
Stomach neoplasms;
Neoplasm metastasis;
Adenovirus;
human;
Protein kinases;
Cyclooxygenase-2
- From:
Chinese Journal of General Surgery
2009;24(9):736-739
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a short hairpin RNA (shRNA) adenovirus vector targeting Aktl (protein kinase B1, PKBI/Aktl) and cyclooxygenase-2 (COX-2) and study its effects on the invasion and metastasis of SGC-7901 human gastric adenocarcinoma cells. Methods Aktl and COX-2 shRNA expression frames were subcloned to pGSadeno adenovirus vector by homologous recombination technology to construct pGSadeno-Aktl+COX-2 (rAdS-A+C) vector. After screening and amplification, the recombinant adenovirus vector was digested with PacI and transfected into SGC-7901 cells, the titer and transfection efficiency were detected by fluorescent microscopy. Aktl and COX-2 mRNA and protein expression was identified by real-time PCR and Western blot. MMP-2 and MMP-9 contents in control group SGC-7901、rAd5-HK and treatment group rAdS-A+C were detected by ELISA assay and transwell assay analyzed cell invasion and metastasis ability. Results Adenovirus vector rAdS-A+C was successfully constructed and it dramatically down-regulated Aktl and COX-2 mRNA and protein expression in SGC-7901 gastric cancer cells. MMP-2 and MMP-9 contents in treatment group rAd5-A+C were respectively (39.7± 1.7) ng/ml, (31.3±3.6) ng/ml, and they were lower than those in control group SGC-7901 (278.4± 15.5) ng/ml, (225.4±15.1) ng/ml and rAd5-HK (275.5±2.1) ng/ml, (226.0±23.3) ng/ml (P= 0.01, P=0.021). Transwell assay showed treatment group rAd5-A+C significantly inhibited the invasion and metastasis of SGC-7901 gastric adenocacinoma cells. Conclusions Adenovirus-mediated targeting Aktl and COX-2 shRNA can inhibit the invasion and metastasis of SGC-7901 human gastric adenocarcinoma cells.
