CAJ | 학술논문

目的：探讨 N-甲基-N′-硝基-N-亚硝基胍（MNNG）对哈萨克族（哈族）正常食管上皮细胞 LRRFIP1、ALDH1L1基因甲基化及其表达的影响。方法将体外培养的哈族正常食管上皮细胞暴露于含0、0．75、1．5、3μg／mL MNNG 的培养基中24 h，分别为对照组、0．75μg／mL 组、1．50μg／mL 组、3．00μg／mL 组。用甲基化特异性聚合酶链（MSP）法检测 LRRFIP1、ALDH1L1基因甲基化状态，用 RT-PCR 和 Western Blot 法检测 LRRFIP1、ALDH1L1基因 mRNA 和蛋白表达。结果与对照组比较，各浓度组细胞 LRRFIP1基因甲基化状态没有改变；各组细胞 LRRFIP1基因 mRNA 表达水平分别为（1．021±0．237）、（2．828±0．350）、（1．453±0．179）、（1．952±0．223）；与对照组比较，0．75μg／mL 组和3．00μg／mL 组表达均升高，差异均具有统计学意义（P ＜0．05）；与对照组比较，1．50μg／mL 和3．00μg／mL 组 LRRFIP1蛋白表达水平均升高，分别为（4．233±0．048）、（4．519±0．033）、（5．377±0．057），差异有统计学意义（P ＜0．05）；高浓度组细胞 ALDH1L1基因有从部分甲基化向完全甲基化转变的趋势，各组 ALDH1L1基因 mRNA 表达水平分别为（1．023±0．254）、（5．046±0．603）、（2．259±0．030）、（7．616±1．910），与对照组比较各浓度组均升高，差异均具有统计学意义（P ＜0．05）；各组细胞 ALDH1L1蛋白表达水平分别为（0．725±0．014）、（0．913±0．033）、（1．142±0．010）、（1．334±0．047），与对照组比较各浓度组均升高，差异均具有统计学意义（P ＜0．05）。结论 MNNG 可促进哈族正常食管上皮细胞 ALDH1L1基因甲基化及 LRRFIP1、ALDH1L1表达的升高。
목적：탐토 N-갑기-N′-초기-N-아초기고（MNNG）대합살극족（합족）정상식관상피세포 LRRFIP1、ALDH1L1기인갑기화급기표체적영향。방법장체외배양적합족정상식관상피세포폭로우함0、0．75、1．5、3μg／mL MNNG 적배양기중24 h，분별위대조조、0．75μg／mL 조、1．50μg／mL 조、3．00μg／mL 조。용갑기화특이성취합매련（MSP）법검측 LRRFIP1、ALDH1L1기인갑기화상태，용 RT-PCR 화 Western Blot 법검측 LRRFIP1、ALDH1L1기인 mRNA 화단백표체。결과여대조조비교，각농도조세포 LRRFIP1기인갑기화상태몰유개변；각조세포 LRRFIP1기인 mRNA 표체수평분별위（1．021±0．237）、（2．828±0．350）、（1．453±0．179）、（1．952±0．223）；여대조조비교，0．75μg／mL 조화3．00μg／mL 조표체균승고，차이균구유통계학의의（P ＜0．05）；여대조조비교，1．50μg／mL 화3．00μg／mL 조 LRRFIP1단백표체수평균승고，분별위（4．233±0．048）、（4．519±0．033）、（5．377±0．057），차이유통계학의의（P ＜0．05）；고농도조세포 ALDH1L1기인유종부분갑기화향완전갑기화전변적추세，각조 ALDH1L1기인 mRNA 표체수평분별위（1．023±0．254）、（5．046±0．603）、（2．259±0．030）、（7．616±1．910），여대조조비교각농도조균승고，차이균구유통계학의의（P ＜0．05）；각조세포 ALDH1L1단백표체수평분별위（0．725±0．014）、（0．913±0．033）、（1．142±0．010）、（1．334±0．047），여대조조비교각농도조균승고，차이균구유통계학의의（P ＜0．05）。결론 MNNG 가촉진합족정상식관상피세포 ALDH1L1기인갑기화급 LRRFIP1、ALDH1L1표체적승고。
Objective To investigate the effects of MNNG on methylation and expressions of LRRFIP1 and ALDH1L1 on Kazakh normal esophageal epithelial cells.Methods Methylation of LRRFIP1 and AL-DH1L1 were detected by MSP,the mRNA and protein level of LRRFIP1 and ALDH1L1 were detected by RT-PCR and Western Blot.Results Compared with the control group,each group of cells with LR-RFIP1 gene methylation status did not change,the expressions of LRRFIP1 mRNA level in each groups were (1.021±0.237),(2.828 ±0.350),(1.453±0.179),(1.952±0.223),the LRRFIP1 mRNA level in low concentration group and high concentration group of cells were significantly higher than the control group (P < 0.05),the expressions of LRRFIP1 protein level in control group and high concentration group were (4.233±0.048),(5.377±0.057),the LRRFIP1 protein level in high concentration group of cells were significantly higher than the control group (P <0.05),high concentration group of cells with ALDH1L1 gene was trended from partial methylation to completely methylation,the expressions of AL-DH1L1 mRNA level in each groups were (1.023±0.254),(5.046±0.603),(2.259 ±0.030),(7.616 ± 1.910),the ALDH1L1 mRNA level in each group was significantly higher than the control group (P <0.05),the expressions of ALDH1L1 protein level in each groups were (0.725±0.014),(0.913±0.033), (1.142±0.010),(1.334±0.047),the ALDH1L1 protein level in each concentration groups were signifi-cantly increased (P < 0.05).Conclusion At some concentration point,MNNG significantly affects the methylation of ALDH1L1 and the expressions of p16 and FHIT on Kazakh normal esophageal epithelial cells.