裸鼠荧光原位移植非小细胞肺癌模型转移相关microRNAs的研究

Metastasis-associated microRNAs in non-small cell lung cancer using fluorescent orthotopic implantation mouse modle

目的:筛选与非小细胞肺癌(NSCLC)转移相关的microRNAs,为研究其在转移过程中发挥的作用提供理论基础。方法:用荧光转染的人NSCLC细胞株H460皮下成瘤后原位移植于裸鼠左肺获取原发灶与转移灶组织标本。通过mi-croRNA芯片检测和RT-PCR验证筛选与转移相关的差异microRNAs,并对其做生物信息学分析。结果:NSCLC有转移原发灶比无转移原发灶(A组)上调的microR-NAs共17个,下调的7个;转移灶比转移原发灶(B组)上调的microRNAs共20个,下调的16个。A组中表达上调的有miR-10b和miR-144,下调的有miR-9、miR-31和miR-34b;B组中表达下调的有miR-25、miR-92a、miR-202和miR-326,它们可能受转录因子AP-1、p53、STATs和NF-κB等调控,作用于RARβ、RASSF1和E2F-1等靶基因,发挥调控细胞增殖周期、细胞发育过程、DNA和RNA代谢及信号传导通路的作用,促进肿瘤生长和转移。结论:在NSCLC有转移和无转移组织中microRNA的表达谱存在差异,它们通过多种途径对肿瘤细胞的增殖、黏附、迁移和浸润起调控作用。

OBJECTIVE：To identify the different expression of metastasis-associated microRNAs in non-small cell lung cancer（NSCLC） for further study on the fuction of them.METHODS：The plasmid was transfected into human NSCLC cell line H460 to get the green fluorescent protein （GFP）-expressing cells.The nude mice were performed with orthotopic transplantation of GFP-H460 cells and killed to obtain the primary and metastasis tumors.The microRNAs were screened by using microarray and RT-PCR,and analyzeed by bioinformatics.RESULTS：The result of microarray showed that there were 17 microRNAs up-regulated and 7 microRNAs down-regulated in the group of primary tissue with metastasis vs primary tissue with non-metastasis （A group）;20 microRNAs up-regulated and 16 microRNAs down-regulated in the group of metastasis tissue vs primary tissue with metastasis （B group）.MiR-10b,miR-144 etc.were up-regulated and miR-9,miR-31,miR-34b etc.were down-regulated in A group,while miR-25,miR-92a,miR-202 and miR-326 etc.were down-regulated in B group.These microRNAs probably were controlled by the transcription factors,such as AP-1,p53,STATs,NF-κB etc.,thus promoted the tumor progression and invasion by targeting RARβ,RASSF1,E2F-1 and so on.CONCLUSIONS：There are differences of the expression of microRNAs between the metastasis and non-metastasis NSCLC tissues.These microRNAs play an important role in tumor proliferation,adhesion,transfer,invasion and metastasis.