中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2010年
7期
622-625
,共4页
杨淋清%纪卫东%陶功华%张文娟%龚春梅%周丽%刘建军%柯跃斌%庄志雄
楊淋清%紀衛東%陶功華%張文娟%龔春梅%週麗%劉建軍%柯躍斌%莊誌雄
양림청%기위동%도공화%장문연%공춘매%주려%류건군%가약빈%장지웅
DNA甲基化%荧光免疫测定%细胞转化,肿瘤%硫化物%镍
DNA甲基化%熒光免疫測定%細胞轉化,腫瘤%硫化物%鎳
DNA갑기화%형광면역측정%세포전화,종류%류화물%얼
DNA methylation%Fluoroimmunoassay%Cell transformation,neoplastic%Sulfides%Nickel
目的 探讨结晶型硫化镍(NiS)对体外培养细胞基因组总体DNA甲基化水平的影响.方法 分别以0.25、0.50、1.00、2.00 μg/cm2结晶型NiS处理人支气管上皮细胞系(16HBE)24 h,隔天再次进行相同处理,共处理3次;以3μmol/L DNA甲基化转移酶抑制剂5-脱氧杂氮胞苷(DAC)处理72 h的正常细胞为阳性对照.应用免疫荧光法和SssI甲基转移酶法定性、定量分析结晶型NiS处理细胞和恶性转化细胞(NSTC)基因组DNA总体甲基化的变化.结果 结晶型NiS处理细胞DNA甲基化免疫荧光的强度均有不同程度降低,转化细胞DNA甲基化免疫荧光的强度明显降低.定量分析结果显示,对照组基因组总体甲基化率(mCpG%)为(81.9±7.3)%,0.25、0.50、1.00、2.00 μg/cm2的结晶型NiS处理3次的细胞(NiS0.25、NiS0.50、NiS1.00、NiS2.00)基因组总体mCpG%则分别为(77.9±6.2)%、(75.3±6.8)%、(59.5±4.9)%、(67.4±5.1)%.经单因素方差分析显示,不同组间总体mCpG%差异有统计学意义(F=124.95,P<0.01),两两比较发现NiS1.00和NiS2.00组与对照组相比,差异均有统计学意义(t值分别为7.64、4.89,P值均<0.01);恶性转化细胞(NSTC1、NSTC2)基因组总体mCpG%分别为(46.2 ±4.1)%、(43.6±4.3)%,低于对照组,差异有统计学意义(t值分别为12.79、13.56,P值均<0.01).结论 结晶型NiS诱发细胞恶性转化过程中,整体基因组DNA甲基化水平降低.
目的 探討結晶型硫化鎳(NiS)對體外培養細胞基因組總體DNA甲基化水平的影響.方法 分彆以0.25、0.50、1.00、2.00 μg/cm2結晶型NiS處理人支氣管上皮細胞繫(16HBE)24 h,隔天再次進行相同處理,共處理3次;以3μmol/L DNA甲基化轉移酶抑製劑5-脫氧雜氮胞苷(DAC)處理72 h的正常細胞為暘性對照.應用免疫熒光法和SssI甲基轉移酶法定性、定量分析結晶型NiS處理細胞和噁性轉化細胞(NSTC)基因組DNA總體甲基化的變化.結果 結晶型NiS處理細胞DNA甲基化免疫熒光的彊度均有不同程度降低,轉化細胞DNA甲基化免疫熒光的彊度明顯降低.定量分析結果顯示,對照組基因組總體甲基化率(mCpG%)為(81.9±7.3)%,0.25、0.50、1.00、2.00 μg/cm2的結晶型NiS處理3次的細胞(NiS0.25、NiS0.50、NiS1.00、NiS2.00)基因組總體mCpG%則分彆為(77.9±6.2)%、(75.3±6.8)%、(59.5±4.9)%、(67.4±5.1)%.經單因素方差分析顯示,不同組間總體mCpG%差異有統計學意義(F=124.95,P<0.01),兩兩比較髮現NiS1.00和NiS2.00組與對照組相比,差異均有統計學意義(t值分彆為7.64、4.89,P值均<0.01);噁性轉化細胞(NSTC1、NSTC2)基因組總體mCpG%分彆為(46.2 ±4.1)%、(43.6±4.3)%,低于對照組,差異有統計學意義(t值分彆為12.79、13.56,P值均<0.01).結論 結晶型NiS誘髮細胞噁性轉化過程中,整體基因組DNA甲基化水平降低.
목적 탐토결정형류화얼(NiS)대체외배양세포기인조총체DNA갑기화수평적영향.방법 분별이0.25、0.50、1.00、2.00 μg/cm2결정형NiS처리인지기관상피세포계(16HBE)24 h,격천재차진행상동처리,공처리3차;이3μmol/L DNA갑기화전이매억제제5-탈양잡담포감(DAC)처리72 h적정상세포위양성대조.응용면역형광법화SssI갑기전이매법정성、정량분석결정형NiS처리세포화악성전화세포(NSTC)기인조DNA총체갑기화적변화.결과 결정형NiS처리세포DNA갑기화면역형광적강도균유불동정도강저,전화세포DNA갑기화면역형광적강도명현강저.정량분석결과현시,대조조기인조총체갑기화솔(mCpG%)위(81.9±7.3)%,0.25、0.50、1.00、2.00 μg/cm2적결정형NiS처리3차적세포(NiS0.25、NiS0.50、NiS1.00、NiS2.00)기인조총체mCpG%칙분별위(77.9±6.2)%、(75.3±6.8)%、(59.5±4.9)%、(67.4±5.1)%.경단인소방차분석현시,불동조간총체mCpG%차이유통계학의의(F=124.95,P<0.01),량량비교발현NiS1.00화NiS2.00조여대조조상비,차이균유통계학의의(t치분별위7.64、4.89,P치균<0.01);악성전화세포(NSTC1、NSTC2)기인조총체mCpG%분별위(46.2 ±4.1)%、(43.6±4.3)%,저우대조조,차이유통계학의의(t치분별위12.79、13.56,P치균<0.01).결론 결정형NiS유발세포악성전화과정중,정체기인조DNA갑기화수평강저.
Objective To observe the effect of crystalline NiS on genome DNA methylation profile in in vitro cultured cells.Methods 16HBE Cells were treated with crystalline NiS at 0.25,0.50,1.00 and 2.00 μg/cm2 for 24 h and three times at total.DAC treatment was given at 3 μmol/L for 72 h.5-mC immunofluorescence and SssI emthyltrasferase assay methods were applied to investigate if the hypomethylation of genome DNA involved.Results The results of 5-mC immunofluorescence showed that the fluorescence intensity of NiS-treated cells were decreased in some degree, and transformed cells were decreased dramatically.By the SssI methylase assay, an average of (81.9 ± 7.3 )% methylated CpG were found in negative control cells.By contrast, ( 77.9 ± 6.2) %, ( 75.3 ± 6.8 ) %, ( 59.5 ± 4.9 ) %, ( 67.4 ±5.1 ) % methylated CpG were observed in cells treated with NiS for three times at dosage of 0.25,0.50,1.00and 2.00 μg/cm2 which were abbreviated as NiS0.25, NiS0.50, NiS1.00, NiS2.00 respectively.The ANOVA analysis results showed that there was a significant difference in the 5 groups above ( F = 124.95,P <0.01 ).The results of Dunnett-t test showed that the methylated CpG of both group NiS1.00 and NiS2.00were significantly decreased compared with the negative control group(t values were 7.64,4.89 respectively,P <0.01 ).For methylated CpG, (46.2 ±4.1 ) % and (43.6% ±4.3)% were observed in NiS-transformed cells (NSTC1 and NSTC2) which were dramatically decreased compared with the negative control group(t values were 12.79,13.56 respectively, P < 0.01 ).Conclusion Genomic DNA methylation levels were decreased during NiS induced malignant transformation.