摘要
Abstract
Objective To observe the effects of silencing P-REX2a on the proliferation and sensibility to doxorubicin of gastric cancer cells SGC7901 and to investigate its mechanism .Methods SGC7901 cells were randomly divided into the normal group (NM group), negative control group (NC group), and interference group (siRNA group).The cells in the NM group were cultured as normal , the cells in the NC group were transfected with negative control sequences , and cells in the siRNA group were transfected with P-REX2a siRNA.We detected the mRNA of P-REX2a by real-time fluores-cent quantitative PCR at 24 h after the transfection .MTT was used to observe cell proliferation in each group ( OD570 ) at 24, 48, and 72 h after the transfection.Some other gastric cancer SGC7901 cells were divided into three groups as before , after 48-hour transfection, they were treated with 0.3 mol/L doxorubicin for 24 h, then we measured the apoptotic rate by using flow cytometry and detected the Bcl-2, Bax, phosphatase and tensin homologue deleted on chromosome 10 (PTEN), Akt, and p-Akt protein by using Western blotting .PTEN phosphatase activity detected by phosphorus PIP 3 substrates-based colorimetric method .Results P-REX2a mRNA of the siRNA group was lower than that of NC group and NM group (P<0.05), but the OD570 was higher than that of the NC group and NM group at 24, 48, and 72 h after the transfection (all P<0.05).After adding doxorubicin, the apoptosis rate was higher in the siRNA group than in the NC group and NM group (P<0.05);the Bax protein expression and phosphatase activity of PTEN was higher but the Bcl -2 and p-Akt protein expression was lower in the siRNA group than in the NC group and NM group (all P<0.05).No significant difference was found in the protein expression of PTEN and Akt between the three groups (both P>0.05).Conclusions Silencing P-REX2a can inhibit the proliferation and improve the sensitivity to adriamycin of gastric cancer SGC 7901 cells.This may be due to that silencing P-REX2a can improve the phosphatase activity of PTEN , inhibit the activation of PI3K/Akt pathway and the expression of Bcl-2, promote the expression of Bax , and thus promote the apoptosis .关键词
胃癌/P-REX2a/基因沉默/小干扰RNA/阿霉素/第10号染色体同源缺失性张力蛋白Key words
gastric carcinoma/P-REX2a/gene silencing/siRNA/doxorubicin/phosphatase and tensin homologue deleted on chromosome 10 ( PTEN)分类
医药卫生
