中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
11期
2685-2687
,共3页
吉西他滨%胞质-5'-核苷酸酶-Ⅱ%乳腺癌%化疗耐药性
吉西他濱%胞質-5'-覈苷痠酶-Ⅱ%乳腺癌%化療耐藥性
길서타빈%포질-5'-핵감산매-Ⅱ%유선암%화료내약성
Gemcitabine%Cytosolic 5'-nucleotidase Ⅱ%Breast cancer%Chemoresistance
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)mRNA和蛋白表达的变化,探讨其在乳腺癌化疗耐药中所起的作用.方法 以50 nmol/L吉西他滨干预MCF-7细胞,流式细胞仪测定0、3、7、10、13、16d细胞凋亡比例,并以实时定量聚合酶链反应(Real-time PCR)及Western blot方法测定存活细胞CN-Ⅱ的mRNA及蛋白表达.结果 干预后第0、3、7、10、13、16 d细胞中CN-Ⅱ的mRNA表达水平分别为1.00±0.21、1.06±0.31、1.11 ±0.29、2.15±0.41、2.46±0.72、3.58±1.34,与干预天数呈正相关(r=0.979),蛋白表达水平分别为1.00±0.31、0.99±0.46、1.03 ±0.42、3.56 ±0.71、4.10±1.25、7.02±1.67,与干预天数呈正相关(r=0.971),两者与凋亡细胞比例的变化趋势一致,相对耐药细胞中的CN-ⅡmRNA和蛋白表达水平与非耐药细胞比较,差异有统计学意义(P＜0.05).结论 CN-Ⅱ在乳腺癌化疗时表达明显增强,可能与化疗耐药性的产生有关.
目的 檢測人乳腺癌MCF-7細胞株經吉西他濱處理時胞質-5'-覈苷痠酶-Ⅱ(CN-Ⅱ)mRNA和蛋白錶達的變化,探討其在乳腺癌化療耐藥中所起的作用.方法 以50 nmol/L吉西他濱榦預MCF-7細胞,流式細胞儀測定0、3、7、10、13、16d細胞凋亡比例,併以實時定量聚閤酶鏈反應(Real-time PCR)及Western blot方法測定存活細胞CN-Ⅱ的mRNA及蛋白錶達.結果 榦預後第0、3、7、10、13、16 d細胞中CN-Ⅱ的mRNA錶達水平分彆為1.00±0.21、1.06±0.31、1.11 ±0.29、2.15±0.41、2.46±0.72、3.58±1.34,與榦預天數呈正相關(r=0.979),蛋白錶達水平分彆為1.00±0.31、0.99±0.46、1.03 ±0.42、3.56 ±0.71、4.10±1.25、7.02±1.67,與榦預天數呈正相關(r=0.971),兩者與凋亡細胞比例的變化趨勢一緻,相對耐藥細胞中的CN-ⅡmRNA和蛋白錶達水平與非耐藥細胞比較,差異有統計學意義(P＜0.05).結論 CN-Ⅱ在乳腺癌化療時錶達明顯增彊,可能與化療耐藥性的產生有關.
목적 검측인유선암MCF-7세포주경길서타빈처리시포질-5'-핵감산매-Ⅱ(CN-Ⅱ)mRNA화단백표체적변화,탐토기재유선암화료내약중소기적작용.방법 이50 nmol/L길서타빈간예MCF-7세포,류식세포의측정0、3、7、10、13、16d세포조망비례,병이실시정량취합매련반응(Real-time PCR)급Western blot방법측정존활세포CN-Ⅱ적mRNA급단백표체.결과 간예후제0、3、7、10、13、16 d세포중CN-Ⅱ적mRNA표체수평분별위1.00±0.21、1.06±0.31、1.11 ±0.29、2.15±0.41、2.46±0.72、3.58±1.34,여간예천수정정상관(r=0.979),단백표체수평분별위1.00±0.31、0.99±0.46、1.03 ±0.42、3.56 ±0.71、4.10±1.25、7.02±1.67,여간예천수정정상관(r=0.971),량자여조망세포비례적변화추세일치,상대내약세포중적CN-ⅡmRNA화단백표체수평여비내약세포비교,차이유통계학의의(P＜0.05).결론 CN-Ⅱ재유선암화료시표체명현증강,가능여화료내약성적산생유관.
Objective To investigate the changes of 5'-nucleotidase Ⅱ (CN-]Ⅱ) expression in human breast cancer cell line MCF-7 after induction of gemcitabine and study the effects of gemcitabine on chemoresistance in breast cancer.Methods After MCF-7 cells were incubated with 50 nmol/L gemcitabine for 0, 3, 7, 10, 13 and 16 days, the apoptosis ratio was determined by flow cytometry, and the mRNA and protein levels of CN-Ⅱ in the surival cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively.Results After MCF-7 cells were incubated with 50 nmol/L gemcitabine for 0, 3, 7, 10, 13 and 16 days, the expression of CN-lⅡ was 1.00 ± 0.21, 1.06 ± 0.31, 1.11 ± 0.29, 2.15 ± 0.41, 2.46 ± 0.72, and 3.58 ± 1.34 at mRNA level, which was positively correlated with incubation time (r =0.979) and consistent with the propprtion of apototic cells.The expression of CN-Ⅱ was 1.00 ±0.31, 0.99 ±0.46, 1.03 ±0.42, 3.56 ±0.71, 4.10 ± 1.25, and 7.02 ± 1.67 at protein level, which was positively correlated with incubation time (r =0.971) and consistent with the propprtion of apototic cells.When the drug resistant cells were compared with the non-drug resistant cells, the expression of mRNA and protein of CN-Ⅱ and APE/Ref-1 showed statistically significant difference.(P ＜ 0.05).Conclusion The obviously up-regulated expression of CN-Ⅱ and APE/Ref-1 may be an adaptive response contributing to the overall chemoresistance in breast cancer.
