miR-103/KLF4在肺癌A549细胞及耐药细胞中的表达检测及靶向关系验证

Detection of expressing of miR-103/KLF4 in A549 lung cancer cells and A549/DDP resistant cells and validation of the targeted relationship

目的研究miR-103/KLF4(Krüppel样因子4)在肺癌A549细胞及耐药细胞中的表达及其靶向关系。方法培养肺癌细胞系A549及耐药细胞系A549/DDP(顺铂)。采用噻唑蓝(MTT)检测细胞的存活率,RT-PCR和Western blot等方法测定两细胞株中miR-103/KLF4的表达情况,双荧光素酶报告基因实验对二者的靶向关系进行验证。结果 miR-103在耐药株细胞A549/DDP中存在高表达,KLF4在耐药株细胞A549/DDP中表达明显降低。在肺癌细胞中,miR-103可以直接靶向KLF4-3'非翻译区(3'UTR)。结论在A549/DDP中miR-103可以直接靶向调控KLF4。

Objective To study the expression and targeted relationship of miR-103/KLF4 （ Kruppel like factor） in A549 and resistant cell lines of lung cancer. Methods To culture the A549 cell lines and A549/DDP （cisplatin） resistant cell lines in vitro and detect survival rates by methyl thiazolyl tetrazolium （MTF） method. Real-time polymerase chain reaction （RT-PCR） and Western blotting were used to test the expression of miR-103/KLF4 of these cell lines. Dual-luciferase reporter gene experiment to detect the targeted relationship. Results A higher expression of miR-103 and a obvious lower expression of KLF4 were observed in A549/DDP resistant cell lines. MiR-103 target KLF4-3′ UTR （3′untranslated regions） directly in lung cancer cells. Conclusions In A549/DDP resistant cells, miR-103 can regulate KLF4 at target.