基于TALE技术的哈萨克族食管上皮DNA甲基转移酶1高表达细胞株模型的构建

doi:10.3969/j.issn.1004-616x.2016.06.003

Abstract

Abstract: OBJECTIVE: To construct esophageal epithelial cells which express high levels of DNA methyltransferase 1 (DNMT1). METHODS: Plasmids WV0133,WV0132 and WV0132 were constructed and transfected into esophageal epithelial cells. Transfection of the cells was observed under the inverted fluorescence microscope. Cells that expressed high levels of DNMT1 were selected by puromycin. Expressions of DNMT1 mRNA and protein were determined by using quantitative real-time PCR and Western blot. RESULTS: Compared with the normal cell group, the mRNA expression levels were 1.786 times and 2.721 times in the WV0133- and WV0132-transfected cells, respectively,and their protein expression levels were 2.734 times and 3.100 times,respectively. They were significantly higher than those in the normal cell group (P<0.01). CONCLUSION: An esophageal epithelial cell line which expressed high levels of DNMT1 was successfully constructed. The cell line can be a valuable tool to study mechanisms for esophageal cancer development.

Key words: DNA methyltransferase 1, esophageal epithelial cell, Kazakh, TALEs

CLC Number:

Q7
R73-35+4

CHEN Yan, ZHANG Huixia. Construction of esophageal epithelial cell with high DNA methyltransferase 1 expression[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(6): 428-431,437.

References

[1] Pennathur A,Gibson MK,Jobe BA,et al. Oesophageal carcinoma[J]. Lancet,2013,381(9864):400-412.
[2] Xiang W,Shi JF,Li P,et al. Estimation of cancer cases and deaths attributable to infection in China[J]. Cancer Cause Control,2011,22(8):1153-1161.
[3] Parkin DM,Bray F,Ferlay J,et al. Global cancer statistics, 2002[J]. CA Cancer J Clin,2005,55(2):74-108.
[4] Cui XB,Pang XL,Li S,et al. Elevated expression patterns and tight correlation of the PLCE1 and NF-κB signaling in Kazakh patients with esophageal carcinoma[J]. Med Oncol, 2014,31(1):791.
[5] Cui XB,Chen YZ,Pang XL,et al. Multiple polymorphisms within the PLCE1 are associated with esophageal cancer via promoting the gene expression in a Chinese Kazakh population[J]. Gene,2013,530(2):315-322.
[6] Shen T,Aneas I,Sakabe NJ,et al. Tbx20 regulates a genetic program essential to adult mouse cardiomyocyte function[J]. J Clin Invest,2011,121(12):4640-4654.
[7] Jones PA. Functions of DNA methylation:islands,start sites,gene bodies and beyond[J]. Nat Rev Genet,2012, 13(7):484-492.
[8] Rhee I,Jair KW,Yen RW,et al. CpG methylation is maintained in human cancer cells lacking DNMT1[J]. Nature, 2000,404(6781):1003-1007.
[9] Kim HJ,Kim JH,Chie EK,et al. DNMT (DNA methyltrans-ferase) inhibitors radiosensitize human cancer cells by suppressing DNA repair activity[J]. Radiat Oncol,2012,7:39.
[10] Zhang F,Cong L,Lodato S,et al. Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription[J]. Nat Biotechnol,2011,29(2):149-153.
[11] 黄思语,阿尔孜古丽·吐尔逊,邓彦超,等. 哈萨克族食管上皮细胞的原代培养方法[J]. 新疆医科大学学报,2014, 37(3):277-279.
[12] Carroll SL. Molecular mechanisms promoting the pathogenesis of Schwann cell neoplasms[J]. Acta Neuropathol,2012,123(3): 321-348.
[13] Wainfan E,Poirier LA. Methyl groups in carcinogenesis: effects on DNA methylation and gene expression[J]. Cancer Res,1992,52(Sup7):2071-2077.
[14] Okano M,Bell DW,Haber DA,et al. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development[J]. Cell,1999,99(3):247-257.
[15] Chen T,Li E. Structure and function of eukaryotic DNA methyltransferases[J]. Curr Top Dev Biol,2004,60:55-89.
[16] Manoharan M,Ramachandran K,Soloway MS,et al. Epigenetic targets in the diagnosis and treatment of prostate cancer[J]. Int Braz J Urol,2007,33(1):11-18.

Construction of esophageal epithelial cell with high DNA methyltransferase 1 expression

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