[1]陈柯,李炳坤,许凯,等.稳定低表达DNA甲基转移酶3b基因对膀胱癌细胞生长和凋亡的影响[J].南方医科大学学报,2015,(11):1524.
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稳定低表达DNA甲基转移酶3b基因对膀胱癌细胞生长和凋亡的影响()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2015年11期
页码:
1524
栏目:
出版日期:
2015-11-15

文章信息/Info

Title:
Effect of stable DNA methyltransferase 3bknockdown on proliferation and apoptosis in bladder cancer cells in vitro
作者:
陈柯; 李炳坤; 许凯; 徐啊白; 刘春晓; 郑少波; 徐亚文; 贾晨尧; 刘奇; 李虎林
关键词:
膀胱癌; DNMT3b; 稳定细胞株; 甲基化
Keywords:
bladder cancer;  DNMT3b;  stable cells;  methylation
摘要:
目的探讨稳定低表达DNA 甲基转移酶3b（DNMT3b）基因对人膀胱癌细胞生长和凋亡的影响。方法包装表达
DNMT3b siRNA和阴性对照siRNA的慢病毒，通过慢病毒感染的方法获得稳定低表达DNMT3b的膀胱癌细胞BIU-87及对照
细胞。MTT和流式细胞仪检测DNMT3b稳定低表达对细胞增殖和凋亡的影响；体内裸鼠成瘤实验观察DNMT3b稳定低表达
对肿瘤的抑制效果；荧光定量PCR和Western blot检测DNMT3b稳定低表达对细胞生长和凋亡相关基因表达的影响；甲基化特
异性PCR检测对细胞生长和凋亡相关基因甲基化的影响。结果荧光定量PCR和Western blot检测结果显示，DNMT3b mRNA
和蛋白水平在BIU-87稳定细胞株中稳定低表达；DNMT3b稳定低表达可以抑制BIU-87细胞的生长，并且抑制裸鼠皮下瘤体的
生长；DNMT3b稳定低表达可以促进BIU-87细胞的凋亡；DNMT3b稳定低表达可以增加凋亡及细胞生长相关基因DAPK，Bax
和RASSF1A mRNA和蛋白水平的表达；DNMT3b稳定低表达可以减少凋亡及细胞生长相关基因DAPK，Bax和RASSF1A启
动子区域的甲基化水平。结论DNMT3b稳定低表达可能通过调控凋亡相关基因以及细胞生长相关基因启动子区域的甲基化
水平来影响基因的表达，从而抑制BIU-87细胞的生长，诱导细胞凋亡。
Abstract:
Objective To investigate the effect of stable knockdown of DNA methyltransferase 3b (DNMT3b) on the proliferation
and apoptosis of bladder cancer cells. Methods Lentivirus expressing DNMT3b siRNA or the negative control siRNA was
infected in human bladder cancer BIU-87 cells. MTT assay and flow cytometry were used to detect cell proliferation and
apoptosis, respectively. The inhibitory effect of DNMT3b knockdown on xenograft tumors in nude mice was observed.
Real-time PCR and Western blotting were carried out to investigate the expression level of cell apoptosis related genes.
Methylation specific PCR was used to examine the methylation in the promoter region of the cell apoptosis related genes.
Results The results of real-time PCR and Western blotting showed that DNMT3b mRNA and protein level were stably
knocked down in BIU-87 cells. Stable DNMT3b knockdown suppressed BIU-87 cell growth and the tumor formation ability of
the cells in nude mice. DNMT3b knockdown promoted the apoptosis of BIU-87 cells, increased the mRNA and protein
expression of the cell growth and apoptosis related genes including DAPK, Bax and RASSF1A, and significantly decreased the
methylation of these genes. Conclusion Stable DNMT3b knockdown can affect the methylation of the cell growth and
apoptosis related genes to regulate their expression, which might be a possible mechanism for suppressed cell growth and
enhanced apoptosis of BIU-87 cells.

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更新日期/Last Update: 1900-01-01