CAJ | 학술논문

目的探讨铁调节蛋白质-2(IRP2)在肺癌铁代谢中的调节作用.方法培养肺腺癌A549细胞`随机分成脂质体组(含脂质体20 mg/L)、对照寡核苷酸组和反义寡核苷酸组3组,每组设10个平行样.通过脂质体介导转染,以脂质体组和对照寡核苷酸组为对照.利用逆转录-PCR、Western印迹检测转铁蛋白、转铁蛋白受体、铁蛋白等铁代谢相关基因mRNA及蛋白的表达.结果转染48 h后,转铁蛋白mRNA在3组之间表达差异无统计学意义(F=2.18,P=0.078);反义寡核苷酸组转铁蛋白受体mRNA表达显著低于脂质体组和对照寡核苷酸组(均P<0.05);反义寡核苷酸组铁蛋白mRNA(0.56±0.06)表达高于脂质体组(0.36±0.05)和对照寡核苷酸组(0.39±0.03)(均P<0.05).转染48 h后,反义寡核苷酸组IRP2蛋白表达明显低于脂质体组和对照寡核苷酸组(P<0.05);转铁蛋白在3组之间表达差异无统计学意义(F=2.668,P=0.088);反义寡核苷酸组转铁蛋白受体蛋白表达低于脂质体组和对照寡核苷酸组(P<0.05);反义寡核苷酸组铁蛋白表达高于脂质体组和对照寡核苷酸组(P<0.05).结论 IRP2可能通过蛋白量的改变来影响肺腺癌A549细胞转铁蛋白受体和铁蛋白的表达,调节铁代谢.
목적 탐토철조절단백질-2(IRP2)재폐암철대사중적조절작용.방법 배양폐선암A549세포`수궤분성지질체조(함지질체20 mg/L)、대조과핵감산조화반의과핵감산조3조,매조설10개평행양.통과지질체개도전염,이지질체조화대조과핵감산조위대조.이용역전록-PCR、Western인적검측전철단백、전철단백수체、철단백등철대사상관기인mRNA급단백적표체.결과 전염48 h후,전철단백mRNA재3조지간표체차이무통계학의의(F=2.18,P=0.078);반의과핵감산조전철단백수체mRNA표체현저저우지질체조화대조과핵감산조(균P<0.05);반의과핵감산조철단백mRNA(0.56±0.06)표체고우지질체조(0.36±0.05)화대조과핵감산조(0.39±0.03)(균P<0.05).전염48 h후,반의과핵감산조IRP2단백표체명현저우지질체조화대조과핵감산조(P<0.05);전철단백재3조지간표체차이무통계학의의(F=2.668,P=0.088);반의과핵감산조전철단백수체단백표체저우지질체조화대조과핵감산조(P<0.05);반의과핵감산조철단백표체고우지질체조화대조과핵감산조(P<0.05).결론 IRP2가능통과단백량적개변래영향폐선암A549세포전철단백수체화철단백적표체,조절철대사.
Objective To discuss the regulating mechanism of iron regulatory protein-2 (IRP2) in the iron metabolism of lung cancer. Methods The cultured A549 cells were divided into 3 groups: liposome group (including liposomes 20 mg/L), random oligonucleotide group (SCODN group) and antisense oligonucleotide group (ASODN group). And the liposome-mediated transfection was employed with the liposome and SCODN groups as controls. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to examine the mRNA and protein expressions of iron metabolism-related transferring (Tf), transferrin receptor (TfR) and ferritin (Fn) genes, etc. Results After a 48-hour transfection, the mRNA expression of Tf had no statistically significant difference among three groups (F=2.18, P=0.078); the mRNA expression of TfR in the ASODN group was significantly lower than that in the liposome and SCODN groups (P＜0.05). The expression of Fn mRNA in the ASODN group (0.56±0.06) was higher than that in the liposome (0.36±0.05) and SCODN groups (0.39±0.03) (P＜0.05). After a 48-hour transfection, the IRP2 protein expression of the ASODN group was significantly lower than those of the liposome and SCODN groups (P<0.05). The Tf protein expression was not statistically different in three groups (F=2.67, P=0.088). The TfR protein expression of the ASODN group was lower than those of the liposome and SCODN groups (P<0.05). And the Fn protein expression of the ASODN group was higher than those of the liposome and SCODN groups (P<0.05). Conclusion IRP2 may affect the expressions of TfR and Fn in lung adenocarcinoma A549 cells by changing the amount of protein and regulating the iron metabolism.