CAJ | 학술논문

目的：研究RNA干扰（RNAi）人肺癌A549细胞中期因子（MK）基因表达及对肿瘤细胞凋亡的影响。方法构建含MK siRNA真核表达载体，将其转染到人肺癌A549细胞株中（实验组），另有阴性对照组（转染阴性对照质粒）和空白对照组。应用RT‐PCR的方法检测各组细胞中MK mRNA的表达情况；采用Western blot法检测MK蛋白的表达情况；用流式细胞仪检测各组细胞凋亡情况。结果实验组MK蛋白和mRNA表达都下降，分别下降了89．3％和83．3％。转染实验组A549细胞有自发性凋亡的发生，早期凋亡比阴性对照组和空白组分别提高了15．5倍和9．34倍。实验组caspase‐3活性明显高于阴性对照组和空白对照组（P＜0．05）。结论 RNAi下调人肺癌A549细胞株MK的表达，可诱导肿瘤细胞的自身凋亡。
목적：연구RNA간우（RNAi）인폐암A549세포중기인자（MK）기인표체급대종류세포조망적영향。방법구건함MK siRNA진핵표체재체，장기전염도인폐암A549세포주중（실험조），령유음성대조조（전염음성대조질립）화공백대조조。응용RT‐PCR적방법검측각조세포중MK mRNA적표체정황；채용Western blot법검측MK단백적표체정황；용류식세포의검측각조세포조망정황。결과실험조MK단백화mRNA표체도하강，분별하강료89．3％화83．3％。전염실험조A549세포유자발성조망적발생，조기조망비음성대조조화공백조분별제고료15．5배화9．34배。실험조caspase‐3활성명현고우음성대조조화공백대조조（P＜0．05）。결론 RNAi하조인폐암A549세포주MK적표체，가유도종류세포적자신조망。
Objective To study the effects of midkine(MK) by RNAi on gene expression and apoptosis in A549 cell .Methods Construct MK siRNA expression vector ,and then was transfected that into A549 cells(positive group ,negative group ,blank cell group) .Expression of MK was determined by RT‐PCR and Western blot in each group ;apoptosis was evaluated by flow cytometry . Results The expression of MK protein declined 89 .3% and expression of mRNA declinded 83 .3% in positive group (positive group) .Spontaneous apoptosis was deteted in positive group ,early apoptosis increased by 15 .5 times and 9 .34 times than the nega‐tive control group and blank group .Activities of caspase‐3 was higher than other two groups(P<0 .05) .Conclusion RNAi can in‐hibit expression of MK and induce apoptosis of A549 cells .