外阴鳞状细胞癌患者癌组织p16和DAPK基因组蛋白H3K27三甲基化状态水平研究
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摘要
目的:研究外阴鳞状细胞癌患者癌组织p16和DAPK基因组蛋白H3K27三甲基化状态及mRNA表达水平。方法:采用染色质免疫共沉淀-实时定量聚合酶联反应(Ch IP-qPCR)技术对8例外阴鳞状细胞癌患者癌组织和毗邻正常组织p16和DAPK基因组蛋白H3K27三甲基化状态进行定量分析;随后采用定量反转录聚合酶联反应(qRT-PCR)检测其基因的mRNA表达水平。结果:外阴鳞状细胞癌患者癌组织p16和DAPK基因组蛋白H3K27三甲基化水平增高;其三甲基化水平较毗邻正常组织有显著性差异(P<0.05);进一步表达分析显示p16和DAPK基因其mRNA表达降低。结论:外阴鳞状细胞癌患者癌组织p16和DAPK基因组蛋白H3K27三甲基化水平增高,组蛋白H3K27三甲基化异常参与外阴鳞状细胞癌的发生,极可能成为外阴鳞状细胞癌新的表观治疗靶点。
Objective: To investigate the histone H3lysine 27 trimethylation( H3K27me3) of p16 and DAPK gene in vulvar squamous cell carcinoma( VSCC) tissue and mRNA expression. Methods: Chromatin immunoprecipitation-quantitative PCR was used to detect the histone H3lysine 27 trimethylation status of p16 and DAPK gene in 8 cases of VSCC and the matched normal tissue. Expression analysis by qRT-PCR is performed on the p16 and DAPK gene.Results: Compared to that of normal tissue,p16 and DAPK gene in VSCC patients displayed increased H3K27me3.Increased H3K27me3 in p16 and DAPK gene lead to decreased mRNA expression. Conclusion: There are significant differences in H3K27me3 alterations between VSCC tissue and the matched normal tissue. H3K27me3 abnormalities in p16 and DAPK gene may be related with tumorigenesis of VSCC. Such novel findings may be become potential biomarker or promising target for epigenetic-based VSCC therapies.
Objective: To investigate the histone H3lysine 27 trimethylation( H3K27me3) of p16 and DAPK gene in vulvar squamous cell carcinoma( VSCC) tissue and mRNA expression. Methods: Chromatin immunoprecipitation-quantitative PCR was used to detect the histone H3lysine 27 trimethylation status of p16 and DAPK gene in 8 cases of VSCC and the matched normal tissue. Expression analysis by qRT-PCR is performed on the p16 and DAPK gene.Results: Compared to that of normal tissue,p16 and DAPK gene in VSCC patients displayed increased H3K27me3.Increased H3K27me3 in p16 and DAPK gene lead to decreased mRNA expression. Conclusion: There are significant differences in H3K27me3 alterations between VSCC tissue and the matched normal tissue. H3K27me3 abnormalities in p16 and DAPK gene may be related with tumorigenesis of VSCC. Such novel findings may be become potential biomarker or promising target for epigenetic-based VSCC therapies.
引文
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[2]Livak KJ,Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))method[J].Methods,2001,25(4):402-408.
[3]Lakshminarasimhan R,Liang G.The role of DNA methylation in cancer[J].Adv Exp Med Biol,2016,945:151-172.
[4]Bhat S,Kabekkodu SP,Varghese VK,et al.Aberrant gene-specific DNA methylation signature analysis in cervical cancer[J].Tumour Biol,2017,39(3):1010428317694573.
[5]Zhong Z,Ye Y,Guo W,et al.Relationship between DLK1 gene promoter region DNA methylation and non-small cell lung cancer biological behavior[J].Oncol Lett,2017,13(6):4123-4126.
[6]Sugai T,Yoshida M,Eizuka M,et al.Analysis of the DNA methylation level of cancer-related genes in colorectal cancer and the surrounding normal mucosa[J].Clin Epigenetics,2017,9:55.
[7]Hyun K,Jeon J,Park K,et al.Writing,erasing and reading histone lysine methylations[J].Exp Mol Med,2017,49(4):e324.
[8]Chen Y,Zhu WG.Biological function and regulation of histone and non-histone lysine methylation in response to DNA damage[J].Acta Biochim Biophys Sin(Shanghai),2016,48(7):603-616.
[9]Ngollo M,Lebert A,Daures M,et al.Global analysis of H3K27me3as an epigenetic marker in prostate cancer progression[J].BMC Cancer,2017,17(1):261.
[10]Tang J,Xie Y,Xu X,et al.Bidirectional transcription of Linc00441 and RB1 via H3K27 modification-dependent way promotes hepatocellular carcinoma[J].Cell Death Dis,2017,8(3):e2675.
[11]Song X,Gao T,Wang NL,et al.Selective inhibition of EZH2 by ZLD1039 blocks H3K27 methylation and leads to potent antitumor activity in breast cancer[J].Sci Rep,2016,6:20864.
[12]Yu Q,Liu Y,Zheng X,et al.Histone H3 lysine 4 trimethylation,lysine27 trimethylation,and lysine 27 acetylation contribute to the transcriptional repression of solute carrier family 47 member 2 in renal cell carcinoma[J].Drug Metab Dispos,2017,45(1):109-117.
[13]Nichol JN,Dupéré-Richer D,Ezponda T,et al.H3K27 methylation:A focal point of epigenetic deregulation in cancer[J].Adv Cancer Res,2016,131:59-95.
[14]Kamb A,Gruis NA,Weaver-Feldhaus J,et al.A cell cycle regulator potentially involved in genesis of many tumor types[J].Science,1994,264(5157):436-440.
[15]Don KR,Ramani P,Ramshankar V,et al.Promoter hypermethylation patterns of P16,DAPK and MGMT in oral squamous cell carcinoma:A systematic review and meta-analysis[J].Indian J Dent Res,2014,25(6):797-805.
[16]Ciesielska U,Zatonski T,Nowinska K,et al.Expression of cell cycle-related proteins p16,p27 and ki-67 proliferating marker in laryngeal squamous cell carcinomas and in laryngeal papillomas[J].Anticancer Res,2017,37(5):2407-2415.
[17]Das M,Sharma SK,Sekhon GS,et al.p16 gene silencing along with p53 single-nucleotide polymorphism and risk of esophageal cancer in Northeast India[J].Tumour Biol,2017,39(5):1010428317698384.
[18]Jiang Y,Xu P,Yao D,et al.CD33,CD96 and death associated protein kinase(dapk)expression are associated with the survival rate and/or response to the chemotherapy in the patients with acute myeloid leukemia(AML)[J].Med Sci Monit,2017,23:1725-1732.
[19]Gade P,Kimball AS,Di Nardo AC,et al.Death-associated protein kinase-1 expression and autophagy in chronic lymphocytic leukemia are dependent on activating transcription factor-6 and CCAAT/enhancer-binding protein-β[J].J Biol Chem,2016,291(42):22030-22042.
[20]Bialik S,Kimchi A.DAP-kinase as a target for drug design in cancer and diseases associated with accelerated cell death[J].Semin Cancer Biol,2004,14(4):283-294.