siRNA抑制真核翻译延长因子1A1表达对结肠癌细胞凋亡和药物敏感性的影响

Effect of eEF1A1 inhibition by siRNA on apoptosis and the chemosensitivity of colon carcinoma cells HCT-116

目的研究特异性siRNA抑制结肠癌细胞HCT-116真核翻译延长因子1A1(e EF1A1)基因及其对结肠癌细胞HCT-116凋亡和药物敏感性的影响。方法设计针对人源结肠癌细胞HCT-116 e EF1A1基因的siRNA,转染至对数生长期的结肠癌细胞HCT-116,分别用反转录-聚合酶链反应和蛋白质印迹法鉴定e EF1A1基因沉默的效果,用流式细胞仪检测e EF1A1基因表达抑制对结肠癌细胞HCT-116凋亡的影响,用噻唑蓝法(MTT)检测e EF1A1基因表达抑制后结肠癌细胞HCT-116药物敏感性的变化。结果 e EF1A1靶向siRNA能显著降低结肠癌细胞HCT-116中e EF1A1的表达,e EF1A1表达抑制后结肠癌细胞HCT-116的增值能力显著低于对照组(P<0.05),5-氟尿嘧啶和顺铂对e EF1A1表达抑制的结肠癌细胞HCT-116的半抑制浓度显著低于对照组(P<0.05)。结论 e EF1A1靶向siRNA能有效抑制e EF1A1基因表达,并能协同增强5-氟尿嘧啶和顺铂对结肠癌细胞HCT-116的杀伤作用和诱导凋亡作用。

Objective To investigate the effect of eukaryotic translation elongation factor 1A1（ e EF1A1） inhibition by siRNA on the apoptotic susceptibility, the chemosensitivity of colon carcinoma cells.Methods Four siRNA specifically targeting e EF1A1 were designed and synthesized in vitro and were transfected into colon carcinoma HCT- 116 cells. Reverse transcription- polymerase chain reaction（ RT- PCR）and Western blot assay were used to validate gene- silencing efficiency of e EF1A1. The cell apoptosis was assessed by flow cytometry（ FCM）.Cell growth state and 50% inhibition concentration（ IC50） of5- fluorouracil（ 5- FU） and cisplatin was determined by MTT assay.Results siRNA specific to e EF1A1 can efficiently decrease the expression of e EF1A1 in colon carcinoma HCT- 116 cells. When e EF1A1 was inhibited, the reproductive activity of colon carcinoma HCT- 116 cells was significantly lower than the control groups（ P 0. 05）. The study also showed that IC50 of 5- FU and cisplatin on colon carcinoma HCT- 116 cells treated by siRNA was decreased（ P 〈0. 05）. Conclusion The siRNA targeting e EF1A1 gene can effectively inhibit the expression of e EF1A1 gene,resulting in enhancing the apoptotic susceptibility,the chemosensitivity of 5- FU and cisplatin on colon carcinoma HCT- 116 cells.