肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2008年
10期
654-657
,共4页
徐妍%胡婷%王春芝%林东军%肖若芝%潘祥林%刘加军
徐妍%鬍婷%王春芝%林東軍%肖若芝%潘祥林%劉加軍
서연%호정%왕춘지%림동군%초약지%반상림%류가군
冬凌草甲素%THP-1细胞%细胞凋亡
鼕凌草甲素%THP-1細胞%細胞凋亡
동릉초갑소%THP-1세포%세포조망
Oridonin%THP-1 cell%Apoptosis
目的 探讨冬凌草甲素对急性单核细胞白血病THP-1细胞的诱导凋亡作用及其作用机制.方法 以不同浓度的冬凌草甲素(16～56 μmol/L)作用于体外培养的THP-1细胞0、24、48及72 h,应用MTT法检测细胞生长抑制率,用Annexin V/PI双染法检测细胞凋亡,Hoechst 33258荧光染色法观察细胞凋亡时的形态学变化,用免疫印迹法(Western Blotting)检测Caspase-3及其裂解底物多聚(ADP-核糖)聚合酶PARP[(poly(ADP-ribose)polymerase)]表达水平的变化.结果 32 μmol/L以上的冬凌草甲素可显著抑制细胞的生长及诱导细胞发生凋亡,呈现出明显的量效与时效关系,在Hoechst 33258荧光染色后,细胞出现核浓缩及核碎裂等典型的凋亡特征.Western Blotting检测结果表明,Caspase-3被活化出现相对分子质量为20×103的裂解片段,PARP被裂解后出现相对分子质量为89×103的亚单位片段.结论 冬凌草甲素能显著抑制THP-1细胞的生长并诱导细胞发生凋亡,通过激活Caspase-3途径可能是冬凌草甲素诱导细胞发生凋亡的重要作用机制;这为冬凌草甲素用于白血病的辅助治疗提供了有力的实验依据.
目的 探討鼕凌草甲素對急性單覈細胞白血病THP-1細胞的誘導凋亡作用及其作用機製.方法 以不同濃度的鼕凌草甲素(16～56 μmol/L)作用于體外培養的THP-1細胞0、24、48及72 h,應用MTT法檢測細胞生長抑製率,用Annexin V/PI雙染法檢測細胞凋亡,Hoechst 33258熒光染色法觀察細胞凋亡時的形態學變化,用免疫印跡法(Western Blotting)檢測Caspase-3及其裂解底物多聚(ADP-覈糖)聚閤酶PARP[(poly(ADP-ribose)polymerase)]錶達水平的變化.結果 32 μmol/L以上的鼕凌草甲素可顯著抑製細胞的生長及誘導細胞髮生凋亡,呈現齣明顯的量效與時效關繫,在Hoechst 33258熒光染色後,細胞齣現覈濃縮及覈碎裂等典型的凋亡特徵.Western Blotting檢測結果錶明,Caspase-3被活化齣現相對分子質量為20×103的裂解片段,PARP被裂解後齣現相對分子質量為89×103的亞單位片段.結論 鼕凌草甲素能顯著抑製THP-1細胞的生長併誘導細胞髮生凋亡,通過激活Caspase-3途徑可能是鼕凌草甲素誘導細胞髮生凋亡的重要作用機製;這為鼕凌草甲素用于白血病的輔助治療提供瞭有力的實驗依據.
목적 탐토동릉초갑소대급성단핵세포백혈병THP-1세포적유도조망작용급기작용궤제.방법 이불동농도적동릉초갑소(16～56 μmol/L)작용우체외배양적THP-1세포0、24、48급72 h,응용MTT법검측세포생장억제솔,용Annexin V/PI쌍염법검측세포조망,Hoechst 33258형광염색법관찰세포조망시적형태학변화,용면역인적법(Western Blotting)검측Caspase-3급기렬해저물다취(ADP-핵당)취합매PARP[(poly(ADP-ribose)polymerase)]표체수평적변화.결과 32 μmol/L이상적동릉초갑소가현저억제세포적생장급유도세포발생조망,정현출명현적량효여시효관계,재Hoechst 33258형광염색후,세포출현핵농축급핵쇄렬등전형적조망특정.Western Blotting검측결과표명,Caspase-3피활화출현상대분자질량위20×103적렬해편단,PARP피렬해후출현상대분자질량위89×103적아단위편단.결론 동릉초갑소능현저억제THP-1세포적생장병유도세포발생조망,통과격활Caspase-3도경가능시동릉초갑소유도세포발생조망적중요작용궤제;저위동릉초갑소용우백혈병적보조치료제공료유력적실험의거.
Objective To investigate the apoptosis inducing effects of oridonin on leukemic THP-1 cells and its mechanisms of action. Methods THP-1 cells in culture medium in vitro were given different concentrations of oridonin (16～56 μmol/L) for 24, 48 and 72 h. The inhibitory rate of the cells were measured by MTT assay, apoptotic morphology was observed by Hoechst 33258 staining, and Annexin V/PI staining was used to detect cell apoptosis by flow cytometry (FCM). Caspase-3 and poly (ADP-ribose) polymerase (PARP) expression were detected by Western blotting. Results Oridonin (over 32 μmol/L) could inhibit the growth of THP-1 cells and cause apoptosis remarkably, the suppression was both in time-and dose-dependentmanner. Marked morphological changes of cell apoptosis such as condensation of chromatin was clearly observed by Hoechst 33258 staining, and Annexin V/PI staining showed that apoptotic cells gradually increased after the cells treated with oridinon. Western blotting showed cleavage of the caspase-3 zymogen protein (32×103), with the appearance of its 20×103 subunit, and a cleaved 89×103 fragment of 116×103 PARP was also found. Conclusion Oridonin can inhibit cell growth and induce apoptosis in THP-1 cells via activation of caspase-3. The results indicated that oridonin might be an important potential anti-leukemia reagent.
