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目的探讨miR-544a对肺癌细胞的侵袭和转移能力的影响及其分子机制。方法用miRNA芯片对非小细胞肺癌(NSCLC)细胞系95C和95D进行miRNA谱系分析,并用荧光定量PCR验证其miR-544a表达水平;用Targetscan软件预测miR-544a的靶基因;Transwell实验观察细胞的侵袭转移能力的变化;western blot验证其表达。结果 miRNA芯片及荧光定量PCR结果显示,与95C细胞(1.00±0.00)比较,miR-544a在95D细胞中表达上调(2.95±0.26,t=12.94,P<0.05);Transwell实验结果显示,95C转染miR-544a mimic后增加(32.00±1.00,q=18.67,P<0.01),95D转染miR-544a inhibitor后,侵袭和转移能力降低(11.00±1.00,q=13.67,P<0.01);Targetscan软件预测E-钙粘连素(CDH1)可能为miR-544a的靶基因;western blot显示,95C转染miR-544a mimic后,CDH1表达下调,vimentin表达上调(0.202±0.017,0.574±0.009,q分别为63.86,9.45,P均<0.01);而95D转染miR-544a inhibitor后,CDH1表达上调,而vimentin表达下调(0.769±0.014,0.320±0.020,q分别为18.67,5.99,P均<0.01)。结论 miR-544a可能通过下调CDH1和上调vimentin的表达来促进肺癌细胞的侵袭和转移。
Objective To investigate the effect of miR-544a on invasion and metastasis of lung cancer cells and its molecular mechanism. Methods miRNA profiling of non-small cell lung cancer (NSCLC) cell lines 95C and 95D was performed using miRNA microarray. The miR-544a expression was verified by real-time PCR. Targetscan software was used to predict the target gene of miR-544a. Invasion and metastasis of changes; western blot verify the expression. Results The results of miRNA microarray and real-time PCR showed that miR-544a was up-regulated in 95D cells compared with that of 95C cells (2.95 ± 0.26, t = 12.94, P <0.05) After transfected with miR-544a inhibitor, miR-544a mimic increased miR-544a mimic expression (32.00 ± 1.00, q = 18.67, P <0.01) The protein predicted that E-cadherin (CDH1) may be the target gene of miR-544a. Western blot analysis showed that CDH1 expression was down-regulated and the expression of vimentin was up-regulated after miR-544a mimic transfected by 95C (0.202 ± 0.017,0.574 ± 0.009, q (63.86,9.45, P <0.01). However, the expression of CDH1 was up-regulated and the expression of vimentin was down-regulated in 95D transfected miR-544a inhibitor group (0.769 ± 0.014,0.320 ± 0.020, q = 18.67, . Conclusion miR-544a may promote the invasion and metastasis of lung cancer cells by down-regulating the expression of CDH1 and up-regulating vimentin.