癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2009年
4期
271-275,279
,共6页
姜英%饶凯敏%陈曦%汪倩%刘爱林%袁晶
薑英%饒凱敏%陳晞%汪倩%劉愛林%袁晶
강영%요개민%진희%왕천%류애림%원정
苯并(a)芘%DNA损伤%p63基因%p73基因
苯併(a)芘%DNA損傷%p63基因%p73基因
분병(a)비%DNA손상%p63기인%p73기인
benzo(a)pyrene%DNA damage%p63 gene%p73 gene
背景与目的:研究p63与p73的mRNA表达与BaP致人肺腺癌细胞(H1299)和人支气管上皮细胞(16HBE)DNA损伤的关系.材料与方法:分别用不同浓度BaP(8、16、32、64和128 μmol/L)处理H1299和16HBE两种细胞,在4 h和12 h时,使用相应的生化检测试剂盒分别测定细胞裂解液中MDA的水平和SOD、GSH-Px的活性,用qRT-PCR方法测定处理后细胞的p53、p63、p73、mdm2和mdm4的mRNA水平;用Comet实验评价细胞DNA损伤程度.结果:16、32和64 μmol/L BaP处理4 h时,两种细胞MDA水平显著性升高,SOD和GSH-Px活性显著性下降(P<0.05).用BaP处理H1299和16HBE细胞4 h和12 h时均观察到DNA损伤随浓度增加而加重,且呈剂量-效应关系(P<0.01),mdm2、mdm4 mRNA表达水平升高(P<0.01).不过仅在12 h时p53基因mRNA表达水平较对照组显著增加(P<0.01).在4 h和12 h时点,仅在H1299细胞的p63和p73 mRNA表达增加(P<0.05). 结论:在BaP致p53缺失的H1299细胞的DNA损伤中,BaP可能通过不依赖p53信号通路激活了p63和p73 mRNA的表达.
揹景與目的:研究p63與p73的mRNA錶達與BaP緻人肺腺癌細胞(H1299)和人支氣管上皮細胞(16HBE)DNA損傷的關繫.材料與方法:分彆用不同濃度BaP(8、16、32、64和128 μmol/L)處理H1299和16HBE兩種細胞,在4 h和12 h時,使用相應的生化檢測試劑盒分彆測定細胞裂解液中MDA的水平和SOD、GSH-Px的活性,用qRT-PCR方法測定處理後細胞的p53、p63、p73、mdm2和mdm4的mRNA水平;用Comet實驗評價細胞DNA損傷程度.結果:16、32和64 μmol/L BaP處理4 h時,兩種細胞MDA水平顯著性升高,SOD和GSH-Px活性顯著性下降(P<0.05).用BaP處理H1299和16HBE細胞4 h和12 h時均觀察到DNA損傷隨濃度增加而加重,且呈劑量-效應關繫(P<0.01),mdm2、mdm4 mRNA錶達水平升高(P<0.01).不過僅在12 h時p53基因mRNA錶達水平較對照組顯著增加(P<0.01).在4 h和12 h時點,僅在H1299細胞的p63和p73 mRNA錶達增加(P<0.05). 結論:在BaP緻p53缺失的H1299細胞的DNA損傷中,BaP可能通過不依賴p53信號通路激活瞭p63和p73 mRNA的錶達.
배경여목적:연구p63여p73적mRNA표체여BaP치인폐선암세포(H1299)화인지기관상피세포(16HBE)DNA손상적관계.재료여방법:분별용불동농도BaP(8、16、32、64화128 μmol/L)처리H1299화16HBE량충세포,재4 h화12 h시,사용상응적생화검측시제합분별측정세포렬해액중MDA적수평화SOD、GSH-Px적활성,용qRT-PCR방법측정처리후세포적p53、p63、p73、mdm2화mdm4적mRNA수평;용Comet실험평개세포DNA손상정도.결과:16、32화64 μmol/L BaP처리4 h시,량충세포MDA수평현저성승고,SOD화GSH-Px활성현저성하강(P<0.05).용BaP처리H1299화16HBE세포4 h화12 h시균관찰도DNA손상수농도증가이가중,차정제량-효응관계(P<0.01),mdm2、mdm4 mRNA표체수평승고(P<0.01).불과부재12 h시p53기인mRNA표체수평교대조조현저증가(P<0.01).재4 h화12 h시점,부재H1299세포적p63화p73 mRNA표체증가(P<0.05). 결론:재BaP치p53결실적H1299세포적DNA손상중,BaP가능통과불의뢰p53신호통로격활료p63화p73 mRNA적표체.
BACKGROUND AND AIM: The associations between the changes of p63 and p73 in mRNA levels and the BaP-induced DNA damage in H1299 cells and in 16HBE cells were investigated. MATERIALS AND METHODS: Both H1299 cells and 16HBE cells were treated with BaP at various concentrations (8, 16, 32, 64 and 128 μmol/L) for 4 h and 12 h.At the two time points, the levels of MDA、SOD and GSH-Px were measured using the test kits. The mRNA levels of p53,p63,p73,mdm2 and mdm4 genes were detected by RT-PCR assay. DNA damage in the cells were evaluated by Comet assay. RESULTS: At 4 h, enhanced MDA level was observed (P<0.05) . However, levels of SOD and GSH-Px were significantly decreased(P < 0.05 for both) in both kinds of cells. After 16HBE and HI299 cells were treated for 4 h and 12 h, DNA damage and mRNA expression levels of mdm2 and mdm4 genes increased in a dose-dependent manner(P < 0.01 for all) . But enhanced mRNA expression level of p53 was observed only at 12 h(P<0.01) . In addition, significant up-regulation of p63 and p73 genes in mRNA levels at the two time points were observed in H1299 cells(P < 0.05) . CONCLUSION: The BaP-induced DNA damage may be associated with enhanced mRNA levels of p63 and p73 genes in the p53-null H1299 cells.
