Effects of hFRNK on human colon cancer cell invasion induced by gastrin

Objective To study the effects of hFRNK gene transfected by adenoviral vector on human colon cancer cell invasion induced by gastrin. Methods The subjects were divided into three groups： control, G17 and hFRNK group. The G17 group was treated with 100 μmol/L gastrin 17 for 12 h to induce Colo320WT cells. As to hFRNK group, Colo320WT cells were infected by pAdhFRNK（MOI：100） for 48 h after transient transfection of pCR3. 1-CAR for 48 h and subsequently treated with gastrinl7 for 12 h, and the control group was untreated Colo320WT cells. Expression of phosphorylated FAK （ py397 ） were assayed by western blot. FAK （ py397 ） at lamellipoda was observed with a confocal microscope. The influence of hFRNK on formation of signal complex of FAK-Src-p130Cas-DocklS0 was assayed with coimmunoprecipitation and immunity blotting. Activity of Rac-GTPase was determined by pull down. Results Phosphorylated FAKTyr397 drastically increased with the induction of gastrin. Compared with that of G17 group, FAK （ py397 ） expression decreased and little FAK（ py397 ） was found at lamellipoda, at the same time, the signal complex of FAK-Src-pl30Cas-Dock180 did not form, and the activity of Rac decreased. Conclusion hFRNK gene may block gastrin-induced FAK phosphorylation, prevent formation of the signal complex, FAK-Src-DocklS0-pl30Cas, and may also downregnlate Rac activation. It could be the theoretical basis of prevention and treatment of tumor invasion and metastasis. Key words: Colon cancer;  Invasiveness, neoplasm;  Genes, hFRNK;  Gastrins