LncRNA FQ223609促进血管平滑肌细胞增殖

LncRNA FQ223609 promote vascular smooth muscle cell proliferation

[目的]构建lncRNA FQ223609真核表达载体,并且探讨其促进血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖的调控机制。[方法]通过RT-PCR扩增得到735 bp的lncRNA FQ223609基因序列,以质粒p CMV-CHA为骨架构建FQ223609真核表达载体,通过HindⅢ/XhoⅠ酶切及测序进行鉴定;将FQ223609真核表达载体及其对照空载体p CMV-C-HA分别转染VSMCs,CCK-8分析细胞活力,Western Blot检测脂蛋白酯酶(lipoprotein lipase,LPL)和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达水平。[结果]双酶切及测序鉴定显示插入lncRNA FQ223609 c DNA序列正确; CCK-8结果表明过表达lncRNA FQ223609使VSMCs增殖活力提高22%; Western Blot结果表明LPL和PCNA的表达水平显著上调。[结论]lncRNA FQ223609真核表达载体成功构建; lncRNA FQ223609通过上调LPL的表达,促进VSMCs增殖。

[Objective]To construct the recombinant eukaryotic expression vector of lncRNA FQ223609,and explore the function of lncRNA FQ223609 in the vascular smooth muscle cell(VSMC)proliferation.[Method]The lncRNA FQ223609 gene about 735 bp was amplified by RT-PCR and then cloned into p CMV-C-HA plasmid to construct lncRNA FQ223609 recombinant vector.After confirmed by HindⅢ/XhoⅠdigestion analysis and sequencing,lncRNA FQ223609 and its empty control vector(p CMV-C-HA)were transfected into VSMCs,the cell viability was detected by CCK-8,the protein levels of lipoprotein lipase(LPL)and proliferating cell nuclear antigen(PCNA)were analyzed by Western Blot.[Result]Restriction enzyme digestion and sequencing showed that the inserted lncRNA FQ223609 cDNA was correct;after overexpressing lncRNA FQ223609 in VSMCs,cell viability was increased by 22%,and the protein levels of LPL and PCNA were upregulated significantly.[Conclusion]The lncRNA FQ223609 recombinant vector was successfully constructed;lncRNA FQ223609 could promote the proliferation of VSMCs via upregulating LPL.