与肺癌侵袭转移相关的miR-424-5p靶基因的预测及鉴定
发布时间：2018-10-11 17:43
【摘要】：为研究miR-424-5p参与肺癌侵袭转移的分子机制,筛选与肺癌侵袭转移相关的miR-424-5p靶基因,采用文献调研分析miR-424-5p与肺癌侵袭转移相关的靶基因,并用生物信息学方法筛选miR-424-5p与靶基因3′UTR结合分数较高的靶基因;结合二者的交集筛选出部分靶基因,构建含靶分子3′UTR的荧光素酶报告基因质粒;分别将miR-424-5p模拟物(mimic)与含有靶分子3′UTR的荧光素酶报告基因质粒共转染293T细胞,进行荧光素酶报告基因实验鉴定靶基因。生物信息学分析表明,与肺癌侵袭转移信号通路相关的基因之一是细胞因子信号传导抑制子5(Suppressor of cytokine signaling 5,SOCS5)。在miR-424-5p mimic或阴性对照(negative control,NC)与野生型靶分子SOCS5 3′UTR双荧光素酶报告基因载体共转入293T细胞后,荧光素酶活性值降低至NC组的46%(P0.01)。而miR-424-5p mimic与靶分子3′UTR突变型报告基因载体共转后,荧光素酶的活性与对照组相比几乎无改变。表明SOCS5可能是miR-424-5p的肺癌侵袭转移相关的一个靶分子。
[Abstract]:In order to study the molecular mechanism of miR-424-5p involved in invasion and metastasis of lung cancer and to screen miR-424-5p target genes associated with invasion and metastasis of lung cancer, the target genes associated with miR-424-5p and invasion and metastasis of lung cancer were analyzed by literature. The target genes with high binding fraction of miR-424-5p and target gene 3'UTR were screened by bioinformatics method, and some target genes were screened by combining the intersection of the two genes, and the plasmids of luciferase reporter gene containing target molecule 3'UTR were constructed. The miR-424-5p mimic (mimic) and the luciferase reporter gene plasmid containing target 3'UTR were cotransfected into 293T cells, and the target gene was identified by luciferase reporter gene experiment. Bioinformatics analysis showed that one of the genes associated with the invasion and metastasis signaling pathway of lung cancer was cytokine signal transduction suppressor 5 (Suppressor of cytokine signaling 5 (SOCS5). After miR-424-5p mimic or negative control (negative control,NC) and wild-type target SOCS5 3'UTR double luciferase reporter gene vector were transfected into 293T cells, luciferase activity decreased to 46% of NC group (P0.01). However, the luciferase activity of miR-424-5p mimic was almost unchanged compared with that of the control group. The results suggest that SOCS5 may be a target molecule associated with invasion and metastasis of miR-424-5p lung cancer.
【作者单位】： 火箭军总医院中心实验室;
【基金】：国家自然科学基金(81172130)
【分类号】：R734.2
[Abstract]:In order to study the molecular mechanism of miR-424-5p involved in invasion and metastasis of lung cancer and to screen miR-424-5p target genes associated with invasion and metastasis of lung cancer, the target genes associated with miR-424-5p and invasion and metastasis of lung cancer were analyzed by literature. The target genes with high binding fraction of miR-424-5p and target gene 3'UTR were screened by bioinformatics method, and some target genes were screened by combining the intersection of the two genes, and the plasmids of luciferase reporter gene containing target molecule 3'UTR were constructed. The miR-424-5p mimic (mimic) and the luciferase reporter gene plasmid containing target 3'UTR were cotransfected into 293T cells, and the target gene was identified by luciferase reporter gene experiment. Bioinformatics analysis showed that one of the genes associated with the invasion and metastasis signaling pathway of lung cancer was cytokine signal transduction suppressor 5 (Suppressor of cytokine signaling 5 (SOCS5). After miR-424-5p mimic or negative control (negative control,NC) and wild-type target SOCS5 3'UTR double luciferase reporter gene vector were transfected into 293T cells, luciferase activity decreased to 46% of NC group (P0.01). However, the luciferase activity of miR-424-5p mimic was almost unchanged compared with that of the control group. The results suggest that SOCS5 may be a target molecule associated with invasion and metastasis of miR-424-5p lung cancer.
【作者单位】： 火箭军总医院中心实验室;
【基金】：国家自然科学基金(81172130)
【分类号】：R734.2
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