白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2011年
5期
261-265
,共5页
朱光荣%唐宇宏%刘佳%季建敏%章亚成%季鸥%朱红青%邵化敏%姜鹏君%沈群
硃光榮%唐宇宏%劉佳%季建敏%章亞成%季鷗%硃紅青%邵化敏%薑鵬君%瀋群
주광영%당우굉%류가%계건민%장아성%계구%주홍청%소화민%강붕군%침군
葛根总黄酮%K562细胞%NB4细胞%细胞凋亡
葛根總黃酮%K562細胞%NB4細胞%細胞凋亡
갈근총황동%K562세포%NB4세포%세포조망
Flavonoids of puerarin%K562 cell line%NB4 cell line%Apoptosis
目的 探讨葛根总黄酮(PR)对慢性粒细胞白血病(CML)细胞株K562和急性早幼粒细胞白血病(APL)细胞株NB4细胞增殖及凋亡的影响.方法 采用MTT法检测PR对K562细胞、NB4细胞的增殖抑制率;光学显微镜及荧光显微镜观察细胞形态改变;Hoechest33258荧光染色AnnexinV/PI双染法检测细胞凋亡率;DNA PI染色法分析细胞周期及亚二倍体峰.Western blot分别检测NB4细胞JNK、PARP、bcl-2、Caspase3,K562细胞bcr-abl、p53、bcl-2、Fas/FasL蛋白表达的变化.结果 12.5～200 μg/ml PR均能抑制K562、NB4细胞增殖.光学显微镜及荧光显微镜下观察到核固缩、凋亡小体等典型的细胞凋亡改变;Annexin V+/PI-细胞呈时间-剂量依赖性增加;DNA PI染色法发现细胞亚二倍体比例增加,G1期比例下降、S期比例增加.PR呈时间-剂量依赖性抑制K562细胞、NB4细胞增殖,诱导细胞凋亡.不同浓度PR干预后K562细胞bcr-abl蛋白水平呈浓度依赖性下调(F=18.74,P<0.05),而bcl-2则无明显变化;p53表达呈浓度依赖性上调;Fas/FasL表达无明显变化.NB4细胞JNK、PARP及Caspase 3蛋白表达与PR浓度呈正相关,与凋亡抑制蛋白bcl-2则呈负相关(F=42.32,P<0.05).结论 PR能有效抑制K562、NB4细胞增殖,阻滞细胞周期进程,诱导细胞凋亡,但分子机制不同.提示一定浓度PR具有较广谱的抗白血病效应.
目的 探討葛根總黃酮(PR)對慢性粒細胞白血病(CML)細胞株K562和急性早幼粒細胞白血病(APL)細胞株NB4細胞增殖及凋亡的影響.方法 採用MTT法檢測PR對K562細胞、NB4細胞的增殖抑製率;光學顯微鏡及熒光顯微鏡觀察細胞形態改變;Hoechest33258熒光染色AnnexinV/PI雙染法檢測細胞凋亡率;DNA PI染色法分析細胞週期及亞二倍體峰.Western blot分彆檢測NB4細胞JNK、PARP、bcl-2、Caspase3,K562細胞bcr-abl、p53、bcl-2、Fas/FasL蛋白錶達的變化.結果 12.5～200 μg/ml PR均能抑製K562、NB4細胞增殖.光學顯微鏡及熒光顯微鏡下觀察到覈固縮、凋亡小體等典型的細胞凋亡改變;Annexin V+/PI-細胞呈時間-劑量依賴性增加;DNA PI染色法髮現細胞亞二倍體比例增加,G1期比例下降、S期比例增加.PR呈時間-劑量依賴性抑製K562細胞、NB4細胞增殖,誘導細胞凋亡.不同濃度PR榦預後K562細胞bcr-abl蛋白水平呈濃度依賴性下調(F=18.74,P<0.05),而bcl-2則無明顯變化;p53錶達呈濃度依賴性上調;Fas/FasL錶達無明顯變化.NB4細胞JNK、PARP及Caspase 3蛋白錶達與PR濃度呈正相關,與凋亡抑製蛋白bcl-2則呈負相關(F=42.32,P<0.05).結論 PR能有效抑製K562、NB4細胞增殖,阻滯細胞週期進程,誘導細胞凋亡,但分子機製不同.提示一定濃度PR具有較廣譜的抗白血病效應.
목적 탐토갈근총황동(PR)대만성립세포백혈병(CML)세포주K562화급성조유립세포백혈병(APL)세포주NB4세포증식급조망적영향.방법 채용MTT법검측PR대K562세포、NB4세포적증식억제솔;광학현미경급형광현미경관찰세포형태개변;Hoechest33258형광염색AnnexinV/PI쌍염법검측세포조망솔;DNA PI염색법분석세포주기급아이배체봉.Western blot분별검측NB4세포JNK、PARP、bcl-2、Caspase3,K562세포bcr-abl、p53、bcl-2、Fas/FasL단백표체적변화.결과 12.5～200 μg/ml PR균능억제K562、NB4세포증식.광학현미경급형광현미경하관찰도핵고축、조망소체등전형적세포조망개변;Annexin V+/PI-세포정시간-제량의뢰성증가;DNA PI염색법발현세포아이배체비례증가,G1기비례하강、S기비례증가.PR정시간-제량의뢰성억제K562세포、NB4세포증식,유도세포조망.불동농도PR간예후K562세포bcr-abl단백수평정농도의뢰성하조(F=18.74,P<0.05),이bcl-2칙무명현변화;p53표체정농도의뢰성상조;Fas/FasL표체무명현변화.NB4세포JNK、PARP급Caspase 3단백표체여PR농도정정상관,여조망억제단백bcl-2칙정부상관(F=42.32,P<0.05).결론 PR능유효억제K562、NB4세포증식,조체세포주기진정,유도세포조망,단분자궤제불동.제시일정농도PR구유교엄보적항백혈병효응.
Objective To explore the effects and the possible molecular mechanism of flavonoids of puerarin (PR) on chronic myelogenous leukemia (CML) cell line K562 and acute promyelocytic leukemia (APL) cell line NB4 in vitro. Methods MTT assays were used to detect the inhibitory effects of cell proliferation. The apoptosis of K562 and NB4 cells was detected by flow cytometry marked with Annexin V/PI. The expression of bcr-abl, p53, bcl-2, Fas/FasL in K562 cells and JNK, PARP, bcl-2 and Caspase 3 in NB4 cells at protein level was detected by Western blot. Results PR could inhibit the proliferation of K562 and NB4 cells in a time-dose dependent manner. The expression of protein levels of bcr-abl fusion gene declined, while the p53 protein otherwise increased, and both were in a dose-dependent manner (F = 18.74, P <0.05). The application of PR had no effect on bcl-2 and Fas/FasL protein expression in K562 cells. The JNK, PARP and Caspase3 proteins were upregulated in NB4 cells, while bcl-2 was downregulated with the increasing concentrations of PR (F=42.32, P <0.05). Conclusion PR could inhibit leukemic cell proliferation, induce cell cycle block, and increase cell apoptosis through different molecular mechanisms. It suggestes that PR might potentially be a kind of broad spectrum anti-leukemia agent.
