與肺癌侵襲轉移相關的miR-424-5p靶基因的預測及鑒定
發(fā)布時間：2018-10-11 17:43
【摘要】：為研究miR-424-5p參與肺癌侵襲轉移的分子機制,篩選與肺癌侵襲轉移相關的miR-424-5p靶基因,采用文獻調(diào)研分析miR-424-5p與肺癌侵襲轉移相關的靶基因,并用生物信息學方法篩選miR-424-5p與靶基因3′UTR結合分數(shù)較高的靶基因;結合二者的交集篩選出部分靶基因,構建含靶分子3′UTR的熒光素酶報告基因質(zhì)粒;分別將miR-424-5p模擬物(mimic)與含有靶分子3′UTR的熒光素酶報告基因質(zhì)粒共轉染293T細胞,進行熒光素酶報告基因?qū)嶒炶b定靶基因。生物信息學分析表明,與肺癌侵襲轉移信號通路相關的基因之一是細胞因子信號傳導抑制子5(Suppressor of cytokine signaling 5,SOCS5)。在miR-424-5p mimic或陰性對照(negative control,NC)與野生型靶分子SOCS5 3′UTR雙熒光素酶報告基因載體共轉入293T細胞后,熒光素酶活性值降低至NC組的46%(P0.01)。而miR-424-5p mimic與靶分子3′UTR突變型報告基因載體共轉后,熒光素酶的活性與對照組相比幾乎無改變。表明SOCS5可能是miR-424-5p的肺癌侵襲轉移相關的一個靶分子。
[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.
【作者單位】： 火箭軍總醫(yī)院中心實驗室;
【基金】：國家自然科學基金(81172130)
【分類號】：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.
【作者單位】： 火箭軍總醫(yī)院中心實驗室;
【基金】：國家自然科學基金(81172130)
【分類號】：R734.2
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