Effect of overexpression of DNA methyltransferase 3B4 gene on proliferation of 293A cells

邹明新; 姜树原; 张姝; 闫少春; 邵国; 刘晓光

doi:10.3969/j.issn.1007-3969.2014.02.004

您当前的位置：

首页 >

文章列表页 >

Effect of overexpression of DNA methyltransferase 3B4 gene on proliferation of 293A cells

更新时间：2025-12-31

- Effect of overexpression of DNA methyltransferase 3B4 gene on proliferation of 293A cells
- China Oncology Vol. 24, Issue 2, Pages: 99-105(2014)
- 作者机构：
  
  1. 包头师范学院生命科学与技术学院,内蒙古,包头,014030
  2. 包头医学院生物医学研究中心及基础医学院,内蒙古,包头,014060
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.1007-3969.2014.02.004
  CLC：
- Published Online：07 March 2014，
  
  Published：07 March 2014
- 稿件说明：
移动端阅览
邹明新,姜树原,张姝,闫少春,邵国,刘晓光. DNA甲基转移酶3B4过表达对293A细胞增殖影响及其机制的探讨[J]. 中国癌症杂志, 2014, 24(2): 99-105.

邹明新, 姜树原, 张姝, et al. Effect of overexpression of DNA methyltransferase 3B4 gene on proliferation of 293A cells[J]. China Oncology, 2014, 24(2): 99-105.
邹明新,姜树原,张姝,闫少春,邵国,刘晓光. DNA甲基转移酶3B4过表达对293A细胞增殖影响及其机制的探讨[J]. 中国癌症杂志, 2014, 24(2): 99-105. DOI： 10.3969/j.issn.1007-3969.2014.02.004.

邹明新, 姜树原, 张姝, et al. Effect of overexpression of DNA methyltransferase 3B4 gene on proliferation of 293A cells[J]. China Oncology, 2014, 24(2): 99-105. DOI： 10.3969/j.issn.1007-3969.2014.02.004.

摘要

背景与目的：DNA甲基转移酶(DNA methyltransferase，DNMT)3B有近40种异构体，它们的表达有组织和疾病的特异性，细胞中这些异构体的功能尚不清楚。本研究旨在探讨外源性DNMT3B4基因过表达对人胚肾细胞株293A增殖的影响及其机制。方法：将携带DNMT3B4基因的质粒pCMV-DNMT3B4及空质粒pCMV-2B转染293A细胞(293A-vector)并形成稳定表达细胞系，培养300 d后用MTT法检测细胞的增殖；流式细胞仪检测细胞的周期分布；real-time PCR蛋白质印迹法(Western blot)检测细胞中p21表达情况；甲基化特异PCR(MS-PCR)检测p21基因启动子区甲基化状态。结果：在接种于96孔板的第4天，2株过表达DNMT3B4的293A细胞(293A-DNMT3B4-1和293A-DNMT3B4-2)吸光度(A)值分别是293A-vector细胞的(58.92±3.47)%和(68.82±5.64)%，过表达DNMT3B4抑制细胞增殖，差异有统计学意义(P0.05)；293A-DNMT3B4-1和293A-DNMT3B4-2的S期细胞比例分别为(35.88±2.00)%和(37.00±1.79)%，293A-vector细胞比例为(40.44±0.91)%，过表达DNMT3B4降低S期细胞比例，差异有统计学意义(P0.05)。过表达DNMT3B4增加p21的表达，但不改变p21基因启动子区甲基化状态。结论：DNMT3B4基因过表达可抑制293A细胞增殖并增加p21表达。

Abstract

Background and purpose: DNMT3B has nearly 40 known splice variants expressed in a tissueand disease-specific manner

but the roles of these splice variants in the cell are still unclear. The aim of this study was to investigate the effects of overexpression of DNA methyltransferase 3B4 (DNMT3B4) gene on proliferation of human embryo kidney 293A cells. Methods: 293A cells were transfected with plasmid pCMV-DNMT3B4 or pCMV-2B and then treated with G418 to get the stable cell line. The stable cell lines were determined for proliferation level by MTT method

and for cell cycle distribution by flow cytometry. The expression of p21 was detected by real-time PCR and Western blot. The methylation status of p21 gene promoter was detected by methylation-specific PCR (MS-PCR). Results: The absorbance value in DNMT3B4-1 and DNMT3B4-2 clone were (58.92±3.47)% and (68.82±5.64)% as compared to 293A-vector cells using MTT method. DNMT3B4 overexpression significantly decreased cell proliferation (P0.05). S phase fraction of 293A-vector cells was (40.44±0.91)%. While in DNMT3B4-1 and DNMT3B4-2 clone cells

the S phase fraction was (35.88±2.00)% and (37.00±1.79)% respectively. Overexpression of DNMT3B4 could significantly decrease S phase fraction (P0.05). The expression of p21 in DNMT3B4 overexpressed cells was increased

but the methylation status of p21 gene promoter was unchanged.Conclusion: Overexpression of DNMT3B4 can inhibit 293A cell proliferation and can facilitate p21 expression.

关键词

Keywords

references

Views

1565

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effects of SEC14L1P1 on proliferation and migration of oral squamous cell carcinoma cells

Study on the mechanism of DDX6 promoting proliferation and migration of nasopharyngeal carcinoma cells by regulating stability of CKMT1A mRNA

Expression of CDC20 in lung adenocarcinoma tissues and its effect on the proliferation and invasion of lung adenocarcinoma cells

Effect of long noncoding RNA FLJ30679 on proliferation and migration of oral squamous cell carcinoma cells

Effect of hsa_circ_0001573 on biological behaviors of breast cancer cells and its molecular mechanism

Related Author

Wentian ZHENG

Hui GONG

Xinyue ZHANG

Jiayi HAO

Yajie WANG

Yingying JIANG

Jiayan XIONG

Wei LEI

Related Institution

School of Stomatology, Shandong Second Medical University

Medical School of Nantong University

Research Institute of Otolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong University

Department of Otolaryngology Head and Neck Surgery, Second Affiliated Hospital of Soochow University

Department of Otolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong University

AI问答

⁰