CAJ | 학술논문

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（eEF1A1）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 eEF1A1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eEF1 A1基因沉默的效果，用流式细胞仪检测eEF1 A1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eEF1A1基因表达抑制后结肠癌细胞HCT－116药物敏感性的变化。结果 eEF1A1靶向siRNA能显著降低结肠癌细胞HCT－116中eEF1A1的表达，eEF1A1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eEF1 A1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 eEF1A1靶向siRNA能有效抑制eEF1A1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。
목적：연구특이성siRNA억제결장암세포HCT－116진핵번역연장인자1A1（eEF1A1）기인급기대결장암세포HCT－116조망화약물민감성적영향。방법설계침대인원결장암세포HCT－116 eEF1A1기인적siRNA，전염지대수생장기적결장암세포HCT－116，분별용반전록－취합매련반응화단백질인적법감정eEF1 A1기인침묵적효과，용류식세포의검측eEF1 A1기인표체억제대결장암세포HCT－116조망적영향，용새서람법（MTT）검측eEF1A1기인표체억제후결장암세포HCT－116약물민감성적변화。결과 eEF1A1파향siRNA능현저강저결장암세포HCT－116중eEF1A1적표체，eEF1A1표체억제후결장암세포HCT－116적증치능력현저저우대조조（P＜0.05），5－불뇨밀정화순박대eEF1 A1표체억제적결장암세포 HCT －116적반억제농도현저저우대조조（P＜0.05）。결론 eEF1A1파향siRNA능유효억제eEF1A1기인표체，병능협동증강5－불뇨밀정화순박대결장암세포HCT－116적살상작용화유도조망작용。
Objective To investigate the effect of eukaryotic translation elongation factor 1A1 (eEF1A1) inhibition by siRNA on the apoptotic susceptibility , the chemosensitivity of colon carcinoma cells.Methods Four siRNA specifically targeting eEF1A1 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 eEF1A1.The cell apoptosis was assessed by flow cytometry ( FCM ).Cell growth state and 50% inhibition concentration ( IC50 ) of 5-fluorouracil ( 5 -FU ) and cisplatin was determined by MTT assay.Results siRNA specific to eEF1A1 can efficiently decrease the expression of eEF1 A1 in colon carcinoma HCT -116 cells.When eEF1A1 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 IC 50 of 5-FU and cisplatin on colon carcinoma HCT -116 cells treated by siRNA was decreased ( P<0.05).Conclusion The siRNA targeting eEF1A1 gene can effectively inhibit the expression of eEF1A1 gene, resulting in enhancing the apoptotic susceptibility , the chemosensitivity of 5-FU and cisplatin on colon carcinoma HCT -116 cells.