天津医科大学学报
天津醫科大學學報
천진의과대학학보
Journal of Tianjin Medical University
2015年
6期
474-479
,共6页
5-aza-2′-deoxycytidine%YTMLC-9%细胞增殖%DNA甲基化%CK13
5-aza-2′-deoxycytidine%YTMLC-9%細胞增殖%DNA甲基化%CK13
5-aza-2′-deoxycytidine%YTMLC-9%세포증식%DNA갑기화%CK13
5-aza-2′-deoxycytidine%human lung squamous-carcinoma YTMLC-9%cell proliferation%DNA methylation%CK13
目的：探讨DNA甲基转移酶抑制剂5-杂氮2′-脱氧胞苷（5-aza-2′-deoxycytidine）对人肺鳞癌YTMLC-9细胞株增殖的影响及对细胞角蛋白13（CK13）表达和CK13基因甲基化的影响。方法：采用四甲基偶氮唑蓝（MTT）法检测不同浓度（1、2、5、10，20、40μmol/L）5-aza-2′-deoxycytidine处理1～4 d后的5-aza-2′-deoxycytidine对人肺鳞癌YTMLC-9细胞株生长的影响；采用半定量PCR及Western blot检测5μmol/L的5-aza-2′-deoxycytidine作用YTMLC-91～4 d CK13的表达；采用甲基化PCR检测不同浓度（1、2、5、10μmol/L）5-aza-2′-deoxycytidine 处理48 h后CK13甲基化和非甲基化的变化。结果：5-aza-2′-deoxycytidine对人肺鳞癌YTMLC-9细胞株的生长具有明显地抑制作用，当用药浓度高于20μmol/L后产生明显的细胞毒性；5-aza-2′-deoxycytidine能够上调CK13基因表达，呈时间依赖性。结论：5-aza-2′-deoxycytidine抑制人肺鳞癌YTMLC-9细胞的生长，抑制作用呈浓度、时间依赖性。5-aza-2′-deoxycytidine可以使CK13重新表达。其生物学效应可能与CK13启动子甲基化状态改变有关。
目的：探討DNA甲基轉移酶抑製劑5-雜氮2′-脫氧胞苷（5-aza-2′-deoxycytidine）對人肺鱗癌YTMLC-9細胞株增殖的影響及對細胞角蛋白13（CK13）錶達和CK13基因甲基化的影響。方法：採用四甲基偶氮唑藍（MTT）法檢測不同濃度（1、2、5、10，20、40μmol/L）5-aza-2′-deoxycytidine處理1～4 d後的5-aza-2′-deoxycytidine對人肺鱗癌YTMLC-9細胞株生長的影響；採用半定量PCR及Western blot檢測5μmol/L的5-aza-2′-deoxycytidine作用YTMLC-91～4 d CK13的錶達；採用甲基化PCR檢測不同濃度（1、2、5、10μmol/L）5-aza-2′-deoxycytidine 處理48 h後CK13甲基化和非甲基化的變化。結果：5-aza-2′-deoxycytidine對人肺鱗癌YTMLC-9細胞株的生長具有明顯地抑製作用，噹用藥濃度高于20μmol/L後產生明顯的細胞毒性；5-aza-2′-deoxycytidine能夠上調CK13基因錶達，呈時間依賴性。結論：5-aza-2′-deoxycytidine抑製人肺鱗癌YTMLC-9細胞的生長，抑製作用呈濃度、時間依賴性。5-aza-2′-deoxycytidine可以使CK13重新錶達。其生物學效應可能與CK13啟動子甲基化狀態改變有關。
목적：탐토DNA갑기전이매억제제5-잡담2′-탈양포감（5-aza-2′-deoxycytidine）대인폐린암YTMLC-9세포주증식적영향급대세포각단백13（CK13）표체화CK13기인갑기화적영향。방법：채용사갑기우담서람（MTT）법검측불동농도（1、2、5、10，20、40μmol/L）5-aza-2′-deoxycytidine처리1～4 d후적5-aza-2′-deoxycytidine대인폐린암YTMLC-9세포주생장적영향；채용반정량PCR급Western blot검측5μmol/L적5-aza-2′-deoxycytidine작용YTMLC-91～4 d CK13적표체；채용갑기화PCR검측불동농도（1、2、5、10μmol/L）5-aza-2′-deoxycytidine 처리48 h후CK13갑기화화비갑기화적변화。결과：5-aza-2′-deoxycytidine대인폐린암YTMLC-9세포주적생장구유명현지억제작용，당용약농도고우20μmol/L후산생명현적세포독성；5-aza-2′-deoxycytidine능구상조CK13기인표체，정시간의뢰성。결론：5-aza-2′-deoxycytidine억제인폐린암YTMLC-9세포적생장，억제작용정농도、시간의뢰성。5-aza-2′-deoxycytidine가이사CK13중신표체。기생물학효응가능여CK13계동자갑기화상태개변유관。
O bjective:To explore the effect of the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine on human lung squamous-carcinoma YTMLC-9 cell lines and the methylation of CK13. Methods:Human lung squamous-carcinoma YTMLC-9 cell lines were treated for 1 to 4 days by 5-aza-2′-deoxycytidine with 1, 2, 5, 10, 20, 40μmol/L, respectively. Consequently, the inhibition rate of the cells were detected by MTT assay, and morphological pattern were observed. CK13 mRNA and protein levels were measured by semi-quantitative RT-PCR and Western blot, respectively, after cells were treated with 5μmol/L 5-aza-2′-deoxycytidine for one to four days. The methylation level of CK13 were detected by methylation-specific polymerase chain reaction (MSP). Results:MTT results showed that the proliferation ability of YTMLC-9 cells was inhibited by 5-aza-2′-deoxycytidine from 1 μmol/L to 10 μmol/L. Furthermore, concentration-dependent and time-dependent manner were shown by(P<0.05).But higher concentration drug derived cells especially under 40μmol/L, were with higher cytoxicity;CK13 expression was reactivated at mRNA and protein level. Incubation time and dose of 5-aza-2′-deoxycytidine also functioned were discovered as factors for the demethylation status of CK13 promoter DNA and CK13 expression. The methylation of CK13 gene promoter was down-regulated by 5-aza-2′-deoxycytidine in a concentration-dependent manner. Conclusion:The proliferation ability of YTMLC-9 cells can be inhibited by 5-aza-2′-deoxycytidine in a concentration-dependent and time-dependent manner. CK13 expression can be reactivated in cells incubated with 5-aza-2′-deoxycytidine. These may associate with the down-regulation of CK13 gene promoter methylation.
