中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2011年
3期
194-197
,共4页
杨淋清%刘庆成%龚春梅%陶功华%刘建军%胡恭华%黄海燕%王昆鹏%庄志雄
楊淋清%劉慶成%龔春梅%陶功華%劉建軍%鬍恭華%黃海燕%王昆鵬%莊誌雄
양림청%류경성%공춘매%도공화%류건군%호공화%황해연%왕곤붕%장지웅
DNA修饰甲基酶类%DNA甲基化%细胞周期
DNA脩飾甲基酶類%DNA甲基化%細胞週期
DNA수식갑기매류%DNA갑기화%세포주기
DNA modification methylases%DNA methylation%Cell cycle
目的 观察人支气管上皮细胞(16HBE)的DNA甲基转移酶1(DNMT1)基因低表达细胞株细胞周期和基因组整体甲基化水平的改变.方法 用慢病毒介导的RNA干扰方法将4个不同的短发夹RNA片段转染16HBE细胞,并用免疫印迹法(Western blotting)检测该细胞DNMT1蛋白表达水平,用流式细胞仪和5-甲基胞嘧啶(5-mC)免疫荧光法检测该细胞的细胞周期和细胞基因组整体甲基化水平.结果 16HBE-shDNMT1-4细胞株的DNMT1蛋白表达与对照组相比平均降低约44%,差异有统计学意义(P＜0.05),但细胞周期和基因组整体甲基化水平无明显改变.结论 成功建立DNMT1基因低表达的16HB细胞株,其细胞周期和基因组整体甲基化水平无明显改变.
目的 觀察人支氣管上皮細胞(16HBE)的DNA甲基轉移酶1(DNMT1)基因低錶達細胞株細胞週期和基因組整體甲基化水平的改變.方法 用慢病毒介導的RNA榦擾方法將4箇不同的短髮夾RNA片段轉染16HBE細胞,併用免疫印跡法(Western blotting)檢測該細胞DNMT1蛋白錶達水平,用流式細胞儀和5-甲基胞嘧啶(5-mC)免疫熒光法檢測該細胞的細胞週期和細胞基因組整體甲基化水平.結果 16HBE-shDNMT1-4細胞株的DNMT1蛋白錶達與對照組相比平均降低約44%,差異有統計學意義(P＜0.05),但細胞週期和基因組整體甲基化水平無明顯改變.結論 成功建立DNMT1基因低錶達的16HB細胞株,其細胞週期和基因組整體甲基化水平無明顯改變.
목적 관찰인지기관상피세포(16HBE)적DNA갑기전이매1(DNMT1)기인저표체세포주세포주기화기인조정체갑기화수평적개변.방법 용만병독개도적RNA간우방법장4개불동적단발협RNA편단전염16HBE세포,병용면역인적법(Western blotting)검측해세포DNMT1단백표체수평,용류식세포의화5-갑기포밀정(5-mC)면역형광법검측해세포적세포주기화세포기인조정체갑기화수평.결과 16HBE-shDNMT1-4세포주적DNMT1단백표체여대조조상비평균강저약44%,차이유통계학의의(P＜0.05),단세포주기화기인조정체갑기화수평무명현개변.결론 성공건립DNMT1기인저표체적16HB세포주,기세포주기화기인조정체갑기화수평무명현개변.
Objective To construct DNA methyltransferase 1 (DNMT1) low expression 16HBE cell line and observe the variation of cell cycle and global genomic DNA methylation. Methods The method of Lenti-virus induced RNA interference was applied to introduce four different shRNA fragment into 16HBE cells. Flow cytometry and 5-mC immunofluorescence methods were used to observe the cell cycle and global DNA methylation status of DNMT1 low expression 16HBE cells. Results The DNMT1 protein relative expression level of 16HBE-shDNMT1-4 cell line was down regulated about 44%(P＜0.05 ) compared with the control. No obvious differences of cell cycle and global genome DNA methylation status were observed between the 16HBE and 16HBE-shDNMT1. Conclusion The DNMT1 gene low expression cell is successfully constructed, and there are no obvious changes happened on the cell cycle and global genomic DNA methylation.
