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目的观察海兔属Ras同源物I(ARHI)对SW480人结肠癌细胞凋亡的影响。方法采用pc DNA3.1-FLAG-ARHI质粒转染和G418筛选法建立稳定表达ARHI的SW480细胞,采用反转录PCR验证ARHI mRNA和蛋白的表达。流式细胞术检测过表达ARHI对结肠癌细胞凋亡的影响;Western blot法检测蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、p53和Bcl-2的蛋白水平。结果成功建立过表达ARHI的SW480细胞,过表达ARHI的SW480细胞凋亡率明显增加,Bcl-2和p-Akt蛋白水平明显降低、P53蛋白表达水平增加,而Akt蛋白水平无明显变化。结论过表达ARHI通过抑制Akt水平增加SW480人结肠癌细胞凋亡。
Objective To observe the effect of Haitian Ras homolog I (ARHI) on the apoptosis of human colorectal cancer cell line SW480. Methods SW480 cells stably expressing ARHI were established by pcDNA3.1-FLAG-ARHI plasmid transfection and G418 selection. ARHI mRNA and protein expression was verified by reverse transcription PCR. Flow cytometry was used to detect the effect of ARHI overexpression on the apoptosis of colon cancer cells. Western blot was used to detect the protein levels of Akt, p-Akt, p53 and Bcl-2. Results ARHI-overexpressing SW480 cells were successfully established. The apoptosis rate of SW480 cells overexpressing ARHI was significantly increased, the protein levels of Bcl-2 and p-Akt were significantly decreased, the expression of P53 protein was increased, while the Akt protein level was unchanged. Conclusion Overexpression of ARHI can increase apoptosis of SW480 human colon cancer cells by inhibiting Akt level.