癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2014年
5期
353-356,360
,共5页
哈萨克族%食管上皮细胞%HIT
哈薩剋族%食管上皮細胞%HIT
합살극족%식관상피세포%HIT
Kazakhs%esophageal epithelial cells%p16%FHIT
目的：探讨N-甲基-N'-硝基-N-亚硝基胍(MNNG)对哈萨克族人群正常食管上皮细胞p16和脆性组氨酸三联体(FHIT)基因甲基化状态及其表达的影响。方法：体外培养哈萨克族人正常食管上皮细胞，用含MNNG浓度分别为0.75、1.50和3.00μg/mL的培养基培养24 h，同时设立空白对照组。采用甲基化特异性聚合酶链(MSP)法检测p16和FHIT基因甲基化状态，实时荧光定量PCR(real time PCR)、Western blot法分别检测p16和FHIT mRNA及蛋白表达水平。结果：与对照组比较，各浓度MNNG处理组细胞p16和FHIT基因甲基化状态均保持未甲基化状态不变。各组细胞p16 mRNA和蛋白表达水平与对照组比较均升高，差异均具有统计学意义(P<0.05或P<0.01)；0.75μg/mL MNNG处理组细胞理组细胞FHIT mRNA表达水平高于对照组(P<0.05)，且3.00μg/mL MNNG处理组FHIT蛋白表达水平较对照组明显升高，差异有统计学意义(P<0.05)。结论：MNNG可促进哈族人群正常食管上皮细胞p16、FHIT mRNA和蛋白的表达。
目的：探討N-甲基-N'-硝基-N-亞硝基胍(MNNG)對哈薩剋族人群正常食管上皮細胞p16和脆性組氨痠三聯體(FHIT)基因甲基化狀態及其錶達的影響。方法：體外培養哈薩剋族人正常食管上皮細胞，用含MNNG濃度分彆為0.75、1.50和3.00μg/mL的培養基培養24 h，同時設立空白對照組。採用甲基化特異性聚閤酶鏈(MSP)法檢測p16和FHIT基因甲基化狀態，實時熒光定量PCR(real time PCR)、Western blot法分彆檢測p16和FHIT mRNA及蛋白錶達水平。結果：與對照組比較，各濃度MNNG處理組細胞p16和FHIT基因甲基化狀態均保持未甲基化狀態不變。各組細胞p16 mRNA和蛋白錶達水平與對照組比較均升高，差異均具有統計學意義(P<0.05或P<0.01)；0.75μg/mL MNNG處理組細胞理組細胞FHIT mRNA錶達水平高于對照組(P<0.05)，且3.00μg/mL MNNG處理組FHIT蛋白錶達水平較對照組明顯升高，差異有統計學意義(P<0.05)。結論：MNNG可促進哈族人群正常食管上皮細胞p16、FHIT mRNA和蛋白的錶達。
목적：탐토N-갑기-N'-초기-N-아초기고(MNNG)대합살극족인군정상식관상피세포p16화취성조안산삼련체(FHIT)기인갑기화상태급기표체적영향。방법：체외배양합살극족인정상식관상피세포，용함MNNG농도분별위0.75、1.50화3.00μg/mL적배양기배양24 h，동시설립공백대조조。채용갑기화특이성취합매련(MSP)법검측p16화FHIT기인갑기화상태，실시형광정량PCR(real time PCR)、Western blot법분별검측p16화FHIT mRNA급단백표체수평。결과：여대조조비교，각농도MNNG처리조세포p16화FHIT기인갑기화상태균보지미갑기화상태불변。각조세포p16 mRNA화단백표체수평여대조조비교균승고，차이균구유통계학의의(P<0.05혹P<0.01)；0.75μg/mL MNNG처리조세포리조세포FHIT mRNA표체수평고우대조조(P<0.05)，차3.00μg/mL MNNG처리조FHIT단백표체수평교대조조명현승고，차이유통계학의의(P<0.05)。결론：MNNG가촉진합족인군정상식관상피세포p16、FHIT mRNA화단백적표체。
OBJECTIVE: To investigate the effects of MNNG on methylation and expressions ofp16 and FHIT genes in normal esophageal epithelial cells in Kazakhs. METHODS:Kazakh people’s normal esophageal epithelial cells were culture in vitro in medium containing MNNG concentrations of 0.00,0.75,1.50,and 3.00 mg/mL for 24 h. Methylation ofp16 andFHIT were analyzed by methylmion specific PCR(MSP),the mRNA and protein level ofp16 and FHIT were measured by real time PCR (RT-PCR) and Western blot. RESULTS:Compared with the control group,p16 andFHIT gene methylation status did not change in treated groups,but the expressions ofp16 mRNA and protein level in treated groups were significantly higher than the control group(P<0.05 orP<0.01). The expressions ofFHIT mRNA level in 0.75μg/mL group was significantly decreased compared with the control group(P<0.01),but in 3.00μg/mL groupFHIT mRNA and protein levels were significantly increased(P<0.05).CONCLUSION:At certain concentration, MNNG could increase the expressions ofp16 andFHIT in normal esophageal epithelial cells in Kazakhs.
