p53通过调控长链非编码RNA lncRNAp53urlnc对舌鳞癌细胞的影响
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摘要
目的研究长链非编码RNA(lncRNA)与p53的表达之间的关系以及敲低lncRNAp53urlnc对舌鳞癌细胞(SCC3)的影响。方法在SCC3细胞中分别敲低和过表达p53后,分别提取各组RNA和蛋白,Western blot法检测p53蛋白的表达,实时定量PCR(qRT-PCR)技术检测lncRNAp53urlnc的表达。构建shlncRNAp53urlnc载体,包装病毒,并感染SCC3细胞,qRT-PCR检测转染效率,采用MTT法、集落形成试验检测shlncRNAp53urlnc对SCC3细胞生长影响。结果 p53敲低使lncRNAp53urlnc表达明显下调,p53过表达使lncRNAp53urlnc上调;敲低lncRNAp53urlnc表达促进SCC3细胞增殖。结论 p53通过正调控lncRNAp53urlnc抑制SCC3细胞增殖。
Objective To investigate the relationship between the long non-coding RNA( lnc RNA) and the expression of p53,and the effect of knocking down lncRNAp53 urlnc in tongue squamous cell carcinoma( SCC3). Methods The expression of p53 in SCC3 cells was knocked down and overexpressed,and then the RNA and protein of each group was collected. The expression of p53 protein was detected by Western blot and the expression of lncRNAp53 urlnc was detected by real-time quantitative PCR( qRT-PCR). shlncRNAp53 urlnc lentiviral vector was constructed,and then shlncRNAp53 urlnc lentiviral vector was transfected into SCC3 cells,the efficiency of the vector was detected by qRT-PCR. The proliferation of SCC3 cells was dectected by MTT and colony forming assay. Results p53 knockdown reduced the expression of lncRNAp53 urlnc significantly and p53 overexpression significantly increased lncRNAp53 urlnc. Knockdown the expression of lncRNAp53 urlnc can promote the proliferation of SCC3 cell. Conclusion lncRNAp53 urlnc inhibits the proliferation of SCC3 cell,which is positive regulated by p53.
Objective To investigate the relationship between the long non-coding RNA( lnc RNA) and the expression of p53,and the effect of knocking down lncRNAp53 urlnc in tongue squamous cell carcinoma( SCC3). Methods The expression of p53 in SCC3 cells was knocked down and overexpressed,and then the RNA and protein of each group was collected. The expression of p53 protein was detected by Western blot and the expression of lncRNAp53 urlnc was detected by real-time quantitative PCR( qRT-PCR). shlncRNAp53 urlnc lentiviral vector was constructed,and then shlncRNAp53 urlnc lentiviral vector was transfected into SCC3 cells,the efficiency of the vector was detected by qRT-PCR. The proliferation of SCC3 cells was dectected by MTT and colony forming assay. Results p53 knockdown reduced the expression of lncRNAp53 urlnc significantly and p53 overexpression significantly increased lncRNAp53 urlnc. Knockdown the expression of lncRNAp53 urlnc can promote the proliferation of SCC3 cell. Conclusion lncRNAp53 urlnc inhibits the proliferation of SCC3 cell,which is positive regulated by p53.
引文
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[2]Sanchez Y,Huarte M.Long non-coding RNAs:challenges for diagnosis and therapies[J].Nucleic Acid Ther,2013,23(1):15-20.
[3]Guttman M,Amit I,Garber M,et al.Chromatin signature reveals over a thousand highly conserved large non-coding RNAs in mammals[J].Nature,2009,458(7235):223-7.
[4]Appella E,Anderson C W.Post-translational modifications and activation of p53 by genotoxic stresses[J].Eur J Biochem,2001,268(10):2764-72.
[5]Bode A M,Dong Z.Post-translational modification of p53 in tumorigenesis[J].Nat Rev Cancer,2004,4(10):793-805.
[6]Kruse J P,Gu W.Snap Shot:p53 posttranslational modifications[J].Cell,2008,133(5):930.
[7]Hermeking H.MicroRNAs in the p53 network:micromanagement of tumour suppression[J].Nat Rev Cancer,2012,12(9):613-26.
[8]Guttman M,Donagyey J,Carey B W,et al.LincRNAs act in the circuitry controlling pluripotency and differentiation[J].Nature,2011,477(7364):295-300.
[9]Oda K,Arakawa H,Tanaka T,et al.p53AIP1,a potential mediator of p53-dependent apoptosis,and its regulation by Ser-46-phosphorylated p53[J].Cell,2000,102(6):849-62.
[10]Bulavin D V,Saito S,Hollander M C,et al.Phosphorylation of human p53 by p38 kinase coordinates N-terminal phosphorylation and apoptosis in response to UV radiation[J].EMBO J,1999,18(23):6845-54.
[11]Schaukowitch K,Kim T K.Emerging epigenetic mechanisms of long non-coding RNAs[J].Neuroscience,2014,264:25-38.
[12]Kim K,Choi J,Heo K,et al.Isolation and characterization of a novel H1.2 complex that acts as a repressor of p53-mediated transcription[J].J Biol Chem,2008,283(14):9113-26.
[13]Snchez Y,Segura V,Marín-Béjar O,et al.Genome-wide analysis of the human p53 transcriptional network unveils a lncRNA tumour suppressor signature[J].Nat Commun,2014,5:5812.