PI3K p85α蛋白表达缺失联合5-FU诱导大肠癌细胞凋亡的分子机制

The Proapoptotic Mechanism of Combination PI3K p85α Depletion and 5-FU Treatment on Colorectal Cancer Cells

目的:探讨PI3Kp85α蛋白表达缺失联合5-FU对大肠癌SW480细胞促凋亡作用的机制。方法:复苏培养PI3Kp85α干扰组SW480细胞(PI3Kp85ɑ/RNAi-SW480)和SW480对照组细胞,Western blot鉴定干扰效果,采用Western blot方法比较两组细胞中凋亡蛋白Bax、Bcl-6、Bim表达的差异。结果:Western blot结果显示SW480干扰组细胞PI3Kp85α蛋白抑制率为84%,5-FU刺激48h后,PI3Kp85ɑ/RNAi-SW480细胞组凋亡蛋白Bax、Bcl-6、Bim的表达较SW480对照组细胞显著增加(P=0.000)。结论:PI3Kp85α蛋白表达缺失联合5-FU可通过增加促凋亡蛋白的表达促进大肠癌SW480细胞凋亡,为进一步阐明大肠癌细胞恶性增值的分子机制打下基础。

Objective： To investigate the proapoptotic mechanism of combination PI3K p85α depletion via RNA interference and 5-FU treatment on colorectal cancer cells.Methods： PI3Kp85ɑ/RNAi-SW480 cells and SW480 control cells were recovered and cultured in RPMI1640,supplemented with 10% fetal calf serum and 500μg/mL G418.Western blot analysis were used to determine the RNA interference effect and to analyze the expression of apoptotic proteins Bax,Bcl-6 and Bim.Results： Western blot analysis showed that the inhibited rate of PI3K p85α protein was 84%.After treated with 5-FU for 48h,the expression of apoptotic protein Bax,Bcl-6 and Bim were significantly higher in PI3K p85α/RNAi-SW480 cells than in SW480 cells（P=0.000）.Conclusion： Combination PI3K p85α depletion and 5-FU treatment could promoteapoptosis of SW480 cells by increasing the expression of proapoptotic proteins.It may be a foundation to further clarify the malignant growth molecular mechanism of colorectal cancer cells.