胃癌细胞RegⅠ基因第二内含子的调控功能

im To observe regulating function of RegⅠ gene intron 2 in gastric cancer cells. Materials and methods The RegⅠ gene intron 2 was cloned from human white blood cells, gastric cancer cell lines BGC823 and SGC7901, and then luciferase reporter vectors of RegⅠgene intron 2 were constructed, named as pGL4-Human-intron 2, pGL4-BGC-823-intron 2 and pGL4-SGC-7901-intron 2, respectively. After identification by XhoⅠ/Hind Ⅲ digestion and sequencing, the recombinant plasmids were transfected into gastric cancer cell lines BGC-823 and SGC-7901, respectively, and screened with G418. The cells were incubated with gastrin at 1×10-6mol/L final concentration for 12h. Luciferase activities were demonstrated with E1500 Kit and detected by Fluorescence Detector. Results The results of sequencing showed that the inserted sequences were corresponded to those in GenBank reported, except for 15 As from base 517 to base 531 of RegⅠ gene, 13 As in pGL4-Human-intron 2, 12 As in pGL4-BGC-823- intron 2, and 14 As in pGL4-SGC-7901-intron 2. The values of luciferase activity in both of BGC-823 and SGC-7901 cells transfected with recombinant plasmids were significantly higher than those in the cells transfected with pGL4 (P＜0.05); that in the cells transfected with pGL4-BGC-823-intron 2 or pGL4-SGC-7901-intron 2 was significantly lower than that in the cells transfected with pGL4-Human-intron 2 (P＜0.05) . After gastrin incubation, no significant difference was found in both of BGC-823 and SGC-7901 cells (P＞0.05).Conclusions RegⅠ gene intron 2 may has enhancing effect on gene expression in gastric cancer cells. The site of 15 As from base 517 to base 531 of RegⅠ gene may be a polymorphic repeat, and the length of A repeat may be associated with the enhancing effect of RegⅠ gene intron 2. Gastrin has no effect on regulating function of RegⅠ gene intron 2 in gastric cancer cells.