中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2014年
6期
519-523
,共5页
宋艳秋%侯俊杰%康丽花%高素君
宋豔鞦%侯俊傑%康麗花%高素君
송염추%후준걸%강려화%고소군
20(S)-人参皂苷Rg3%多发性骨髓瘤%细胞凋亡%U266细胞系%血管内皮生长因子
20(S)-人參皂苷Rg3%多髮性骨髓瘤%細胞凋亡%U266細胞繫%血管內皮生長因子
20(S)-인삼조감Rg3%다발성골수류%세포조망%U266세포계%혈관내피생장인자
20(S)-ginsenoside Rg3%Multiple myeloma%Apoptosis%U266 cell%Vascular endothelial growth factor
目的 探讨20(S)-人参皂苷Rg3[简称20(S)-Rg3]体外对多发性骨髓瘤(MM)细胞增殖及其自分泌血管内皮生长因子(VEGF)的影响.方法 以MM细胞系U266细胞为研究对象,利用MTT法检测不同浓度20(S)-Rg3对细胞增殖的影响;流式细胞术检测细胞周期及凋亡率;ELISA法检测细胞培养上清VEGF水平;Westem blot法检测凋亡相关蛋白caspase-3、8和9的表达.结果 20(S)-Rg3在一定浓度范围内可抑制U266细胞增殖,并呈浓度依赖性(P＜0.05),作用24、48 h的IC50值分别为(71.07±2.63)μmol/L和(44.06±3.98)μmol/L;ELISA法检测结果显示20(S)-Rg3处理24 h细胞培养上清中VEGF的含量呈剂量依赖性减少(P＜0.05),未加药组和加近IC50浓度(80 μmol/L)组的细胞培养上清中VEGF的含量分别为(419.93±36.76)和(314.82±27.05)pg/106细胞.流式细胞术检测显示0、20、40、80 μmol/L 20(S)-Rg3处理后U266细胞凋亡率分别为(0.51±0.05)％、(8.32±0.83)％、(10.72±1.29)％和(15.27±2.26)％,呈剂量依赖性升高(P＞0.05);G0/G1期细胞比率分别为(49.11±1.71)％、(52.72±7.75)％、(60.29±5.76)％和(61.81±3.46)％,呈剂量依赖性增多(P＜ 0.05);Western blot检测显示随20(S)-Rg3浓度的增加,procaspase-3、8和9表达略下降,cleaved caspase-3、8和9显著表达上升(P＜0.05).结论 20(S)-Rg3可抑制U266细胞增殖,其机制可能是通过诱导细胞凋亡,上调cleaved cas-pase-3、8、9的表达及阻滞细胞周期进程实现的;20(S)-Rg3可抑制U266细胞分泌VEGF,潜在治疗MM的新制剂.
目的 探討20(S)-人參皂苷Rg3[簡稱20(S)-Rg3]體外對多髮性骨髓瘤(MM)細胞增殖及其自分泌血管內皮生長因子(VEGF)的影響.方法 以MM細胞繫U266細胞為研究對象,利用MTT法檢測不同濃度20(S)-Rg3對細胞增殖的影響;流式細胞術檢測細胞週期及凋亡率;ELISA法檢測細胞培養上清VEGF水平;Westem blot法檢測凋亡相關蛋白caspase-3、8和9的錶達.結果 20(S)-Rg3在一定濃度範圍內可抑製U266細胞增殖,併呈濃度依賴性(P＜0.05),作用24、48 h的IC50值分彆為(71.07±2.63)μmol/L和(44.06±3.98)μmol/L;ELISA法檢測結果顯示20(S)-Rg3處理24 h細胞培養上清中VEGF的含量呈劑量依賴性減少(P＜0.05),未加藥組和加近IC50濃度(80 μmol/L)組的細胞培養上清中VEGF的含量分彆為(419.93±36.76)和(314.82±27.05)pg/106細胞.流式細胞術檢測顯示0、20、40、80 μmol/L 20(S)-Rg3處理後U266細胞凋亡率分彆為(0.51±0.05)％、(8.32±0.83)％、(10.72±1.29)％和(15.27±2.26)％,呈劑量依賴性升高(P＞0.05);G0/G1期細胞比率分彆為(49.11±1.71)％、(52.72±7.75)％、(60.29±5.76)％和(61.81±3.46)％,呈劑量依賴性增多(P＜ 0.05);Western blot檢測顯示隨20(S)-Rg3濃度的增加,procaspase-3、8和9錶達略下降,cleaved caspase-3、8和9顯著錶達上升(P＜0.05).結論 20(S)-Rg3可抑製U266細胞增殖,其機製可能是通過誘導細胞凋亡,上調cleaved cas-pase-3、8、9的錶達及阻滯細胞週期進程實現的;20(S)-Rg3可抑製U266細胞分泌VEGF,潛在治療MM的新製劑.
목적 탐토20(S)-인삼조감Rg3[간칭20(S)-Rg3]체외대다발성골수류(MM)세포증식급기자분비혈관내피생장인자(VEGF)적영향.방법 이MM세포계U266세포위연구대상,이용MTT법검측불동농도20(S)-Rg3대세포증식적영향;류식세포술검측세포주기급조망솔;ELISA법검측세포배양상청VEGF수평;Westem blot법검측조망상관단백caspase-3、8화9적표체.결과 20(S)-Rg3재일정농도범위내가억제U266세포증식,병정농도의뢰성(P＜0.05),작용24、48 h적IC50치분별위(71.07±2.63)μmol/L화(44.06±3.98)μmol/L;ELISA법검측결과현시20(S)-Rg3처리24 h세포배양상청중VEGF적함량정제량의뢰성감소(P＜0.05),미가약조화가근IC50농도(80 μmol/L)조적세포배양상청중VEGF적함량분별위(419.93±36.76)화(314.82±27.05)pg/106세포.류식세포술검측현시0、20、40、80 μmol/L 20(S)-Rg3처리후U266세포조망솔분별위(0.51±0.05)％、(8.32±0.83)％、(10.72±1.29)％화(15.27±2.26)％,정제량의뢰성승고(P＞0.05);G0/G1기세포비솔분별위(49.11±1.71)％、(52.72±7.75)％、(60.29±5.76)％화(61.81±3.46)％,정제량의뢰성증다(P＜ 0.05);Western blot검측현시수20(S)-Rg3농도적증가,procaspase-3、8화9표체략하강,cleaved caspase-3、8화9현저표체상승(P＜0.05).결론 20(S)-Rg3가억제U266세포증식,기궤제가능시통과유도세포조망,상조cleaved cas-pase-3、8、9적표체급조체세포주기진정실현적;20(S)-Rg3가억제U266세포분비VEGF,잠재치료MM적신제제.
Objective To explore the effect of 20 (S)-ginsenoside Rg3 [20 (S)-Rg3] on the proliferation inhibition and secretion of vascular endothelial growth factor (VEGF) of multiple myeloma (MM) cell line U266.Methods The proliferation inhibition rate of U266 cells after treatment with different doses of 20 (S)-Rg3 was detected by MTT method,the cell cycle and apoptosis by flow cytometry,the expression of apoptosis related proteins of caspase-3,8 and 9 by Western blot,VEGF concentration in the culture supernatant by ELISA.Results It showed that 20 (S)-Rg3 could inhibit the proliferation of U266 in a dose-dependent manner (P＜0.05) with IC50 of (71.07±2.63) μmol/L and (44.06± 3.98) μmol/L at 24 h and 48 h,respectively.VEGF concentration in the culture supernatant showed a dose-dependent reduction (P＜0.05),decreased from (419.93 ± 36.76) pg/106cells in the control group to (314.82±27.05) pg/106cells in 80 μmol/L 20 (S)-Rg3 treated group by ELISA assay.Flow cytometry with Annexin-V/PI double staining revealed that 20 (S)-Rg3 may induce U266 cells apoptosis in a concentration-dependent manner from (0.51±0.05)％ at control group to (8.32±0.83)％,(10.72±1.29)％ and (15.27±2.26)％ at 20,40 and 80μmol/L treatment groups,respectively (P＜0.05).Flow cytometry with PI staining showed that the ratio of cells in G0/G1 phase increased from (49.11±1.71)％ to (52.72±7.75)％,(60.29± 5.76) ％ and (61.81±3.46) ％,respectively (P＜0.05).Western blot analysis indicated that the expression of caspase-3,8 and 9 declined,and that ofcleaved-caspase-3,8 and 9 significantly increased (P＜0.05) with 20 (S)-Rg3 concentration increased.Conclusion 20(S)-Rg3 can inhibit the proliferation ofU266 cells by cell cycle arrest in Gi phase and induce cell apoptosis by increasing the expressions of cleaved-caspase-3,-8 and-9.It can also inhibit VEGF secretion of U266 cells,which makes it a potential agent for multiple myeloma therapy.
