CAJ | 학술논문

目的检测人乳腺癌MCF-7细胞株经吉西他滨处理时胞质-5'-核苷酸酶-Ⅱ(CN-Ⅱ)、脱嘌呤/脱嘧啶核酸内切酶(APE)/氧化还原因子-1(Ref-1) mRNA和蛋白质表达的变化,探讨两者在乳腺癌化疗耐药中的作用.方法分别以3.125、6.250、12.500、25.000 mg/L吉西他滨诱导MCF-7细胞株24h,以0mg/L吉西他滨做空白对照,应用实时定量聚合酶链反应(Real-time PCR)及Westem blot方法测定用药后CN-Ⅱ、APE/Ref-1的mRNA及蛋白表达.结果吉西他滨作用于人乳腺癌MCF-7细胞株24h后,CN-Ⅱ、APE/Ref-1的mRNA表达水平均明显上升,与吉西他滨的浓度呈正相关,其中6.250、12.500、25.000 mg/L吉西他滨干预后CN-Ⅱ相对表达量分别是1.75±0.31、2.50±0.36、2.87±0.57;APE/Ref-l相对表达水平分别是1.65±0.22、2.08±0.39、2.86±0.44(r =0.997,P＜0.05).蛋白表达与吉西他滨的浓度呈正相关,12.500、25.000 mg/L吉西他滨干预后CN-Ⅱ蛋白相对表达量为1.70±0.37、2.21±0.61;APE/Re f-1蛋白相对表达水平分别是1.95±0.46、2.87±0.29(r =0.989,P＜0.05).结论随着吉西他滨的浓度增加与乳腺癌细胞中APE/Ref-1和CN-Ⅱ表达均升高,为吉西他滨诱导乳腺癌化疗过程中对耐药机制有关.
목적 검측인유선암MCF-7세포주경길서타빈처리시포질-5'-핵감산매-Ⅱ(CN-Ⅱ)、탈표령/탈밀정핵산내절매(APE)/양화환원인자-1(Ref-1) mRNA화단백질표체적변화,탐토량자재유선암화료내약중적작용.방법 분별이3.125、6.250、12.500、25.000 mg/L길서타빈유도MCF-7세포주24h,이0mg/L길서타빈주공백대조,응용실시정량취합매련반응(Real-time PCR)급Westem blot방법측정용약후CN-Ⅱ、APE/Ref-1적mRNA급단백표체.결과 길서타빈작용우인유선암MCF-7세포주24h후,CN-Ⅱ、APE/Ref-1적mRNA표체수평균명현상승,여길서타빈적농도정정상관,기중6.250、12.500、25.000 mg/L길서타빈간예후CN-Ⅱ상대표체량분별시1.75±0.31、2.50±0.36、2.87±0.57;APE/Ref-l상대표체수평분별시1.65±0.22、2.08±0.39、2.86±0.44(r =0.997,P＜0.05).단백표체여길서타빈적농도정정상관,12.500、25.000 mg/L길서타빈간예후CN-Ⅱ단백상대표체량위1.70±0.37、2.21±0.61;APE/Re f-1단백상대표체수평분별시1.95±0.46、2.87±0.29(r =0.989,P＜0.05).결론 수착길서타빈적농도증가여유선암세포중APE/Ref-1화CN-Ⅱ표체균승고,위길서타빈유도유선암화료과정중대내약궤제유관.
Objective To investigate the changeof cytosoli5'-nucleotidase Ⅱ (CN-Ⅱ) and apurinic/apyrimidiniendonuclease (APE)/redox factor-1 (Ref-1) expression in human breascancecell line MCF-7 afteinduction of gemcitabine.MethodMCF-7 cellwere incubated with 3.125, 6.250, 12.500, 25.000 mg/L of gemcitabine respectively fo24 h, and 0 mg/L ablank control.The mRNand protein expression of CN-Ⅱ and APE/Ref-1 wadetected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting, respectively.ResultTwenty-fouh aftetreatmenwith gemcitabine, the expression levelof CN-Ⅱ and APE/Ref-1 mRNand protein in MCF-7 cellwere elevated significantly, CN-Ⅱ expression were 1.75 ±0.31, 2.50 ±0.36, 2.87 ± 0.57 afteincubated with 6.250, 12.500, 25.000 mg/L of gemcitabine,and APE/Ref-1 expression were 1.65 ± 0.22, 2.08 ± 0.39, 2.86 ± 0.44 (=0.997, P ＜ 0.05).The expression of protein of CN-Ⅱ and APE/Ref-1 in drug resistancellhave statistical significance.CN-Ⅱ expression were 1.70 ± 0.37,2.21 ±0.61 afteincubated with 12.500, 25.000 mg/L1 of gemcitabine, and APE/Ref-1 expression were 1.95 ± 0.46, 2.87 ± 0.29 (=0.989, P ＜ 0.05).Conclusion The obviouup-regulated expression of CN-Ⅱ and APE/Ref-1 with increaseof gemcitabine may be an adaptive response contributing to the overall chemoresistance in breascancer.