CAJ | 학술논문

目的探讨唑来膦酸诱导结肠癌HCT116细胞凋亡的线粒体途径.方法分别采用浓度25μmol/L和50 μmol/L的唑来膦酸孵育HCT116细胞48 h,流式细胞仪分析凋亡细胞比例；将标志线粒体膜电位的染料JC-1与唑来膦酸作用24、48、72 h的HCT116细胞避光孵育15 min,荧光显微镜和流式细胞仪观察其红色荧光强度的变化；Western blot分析唑来膦酸作用后48、72 h细胞线粒体和胞质细胞色素C(Cyclin C)含量的改变,细胞线粒体和细胞质Cyclin C的改变.结果 HCT 116细胞和唑来膦酸25μmol/L和50 μmol/L作用48 h后,出现凋亡,凋亡比例分别为(11.6±0.5)％、(49.2±3.4)％(P＜0.01)；唑来膦酸浓度25 μmol/L时,24、48、72 h后JC-1被激发出红色荧光的细胞比例为(96.49±2.10)％、(86.13±3.20)％、(74.23 ±5.30)％ (P ＜0.01)；Western blot显示唑来膦酸作用下,Cyclin C从线粒体内释放至细胞质.结论线粒体途径是唑来膦酸介导的结肠癌HCT116细胞凋亡的信号途径之一.
목적 탐토서래련산유도결장암HCT116세포조망적선립체도경.방법 분별채용농도25μmol/L화50 μmol/L적서래련산부육HCT116세포48 h,류식세포의분석조망세포비례；장표지선립체막전위적염료JC-1여서래련산작용24、48、72 h적HCT116세포피광부육15 min,형광현미경화류식세포의관찰기홍색형광강도적변화；Western blot분석서래련산작용후48、72 h세포선립체화포질세포색소C(Cyclin C)함량적개변,세포선립체화세포질Cyclin C적개변.결과 HCT 116세포화서래련산25μmol/L화50 μmol/L작용48 h후,출현조망,조망비례분별위(11.6±0.5)％、(49.2±3.4)％(P＜0.01)；서래련산농도25 μmol/L시,24、48、72 h후JC-1피격발출홍색형광적세포비례위(96.49±2.10)％、(86.13±3.20)％、(74.23 ±5.30)％ (P ＜0.01)；Western blot현시서래련산작용하,Cyclin C종선립체내석방지세포질.결론 선립체도경시서래련산개도적결장암HCT116세포조망적신호도경지일.
Objective To study the mechanism of zoledronate (ZOL) triggering colorectal cancer cell line HCT116 apoptosis.Methods Flow cytometry detected the apoptosis of colorectal cancer cell line HCT116 treated by ZOL with 25 μmol/L or 50 μmol/L for 48 h; Fluorescent labeling and flow cytometry detected the JC-1 fluorescence changes in mitochondria; Western blotting analyzed the distribution of cytochrome C in mitochondria and cytosol.Results After 48 h of drug treatment,there was a significant apoptosis group in HCT116 cells [(11.6 ± 0.5) ％,25 μmol/L; (49.2 ± 3.4) ％,50 μmol/L; P ＜ 0.01].The decreased fluorescence of JC-1 proved that the decreased mitochondrial membrane potential and dysfunction of mitochondria membrane [(96.49 ± 2.10)％,25 μmol/L,24 h; (86.13 ± 3.20)％,25 μ mol/L,48 h; (74.23 ± 5.30) ％,25 μmol/L,72 h; P ＜ 0.01].Meanwhile,Western blotting analysis showed that cytochrome C was decreased in mitochondria and increased in cytosol.It indicated that the cytochrome C was released from mitochondria after treatment with ZOL.Conclusion ZOL triggers HCTll6 colorectal cancer cell line apoptosis by mitochondria pathway.