肝脏
肝髒
간장
Chinese Hepatology
2015年
10期
786-790
,共5页
张懿%倪鎏达%周丰%杨峻%程明亮%傅青春%陈成伟%吴银霞
張懿%倪鎏達%週豐%楊峻%程明亮%傅青春%陳成偉%吳銀霞
장의%예류체%주봉%양준%정명량%부청춘%진성위%오은하
还原型谷胱甘肽%多柔比星%人肝癌细胞株%HepG2
還原型穀胱甘肽%多柔比星%人肝癌細胞株%HepG2
환원형곡광감태%다유비성%인간암세포주%HepG2
Reduced glutathione%Doxorubicin%Human hepatoma cell line%HepG2
目的:研究还原型谷胱甘肽(GSH)对多柔比星(DOX)处理 HepG2细胞株增殖及凋亡的影响;探索核转录因子(NF)-κB p65、Bcl-2在 DOX 单药及联合 GSH 诱导 HepG2细胞凋亡后的表达及其意义。方法实验分4组,空白组:培养液,对照组:培养液+HepG2细胞+RPMl l640培养基,DOX 组:培养液+HepG2细胞+DOX,DOX+GSH 组:培养液+HepG2细胞+DOX+GSH。MTT 法检测 HepG2细胞生长,流式细胞仪检测细胞早期凋亡率,实时荧光定量 PCR 法检测 NF-κB p65 mRNA 的表达,Western 印迹检测 NF-κB p65及 Bcl-2的表达。结果(1)DOX 组和 DOX+GSH 组作用细胞24 h、48 h、72 h 均能显著抑制 HepG2细胞增殖,DOX 组抑制率显著高于 DOX+GSH 组(P <0.05),且抑制率具有时间和剂量依赖性。(2)对照组凋亡率为(0.733±0.153)%,DOX 组凋亡率为(28.400±0.007)%,DOX+GSH 组凋亡率为(15.500±0.006)%;DOX 组、DOX+GSH 组分别与对照组相比,差异有统计学意义(P <0.01);DOX 组与 DOX+GSH 组相比,差异有统计学意义(P <0.01)。(3)两组在处理细胞24 h 后,DOX+GSH 组 NF-κB p65mRNA 表达显著高于 DOX组(P <0.05)。(4)DOX 组 NF-κB p65、Bcl-2表达较对照组增高,DOX+GSH 组表达较 DOX 组表达增高。结论 GSH 与DOX 联合使用可导致 DOX 化疗效果下降,其机制可能是通过进一步上调 NF-κB p65和 Bcl-2的表达实现的。临床上使用DOX 化疗的肿瘤患者应避免同时使用 GSH。
目的:研究還原型穀胱甘肽(GSH)對多柔比星(DOX)處理 HepG2細胞株增殖及凋亡的影響;探索覈轉錄因子(NF)-κB p65、Bcl-2在 DOX 單藥及聯閤 GSH 誘導 HepG2細胞凋亡後的錶達及其意義。方法實驗分4組,空白組:培養液,對照組:培養液+HepG2細胞+RPMl l640培養基,DOX 組:培養液+HepG2細胞+DOX,DOX+GSH 組:培養液+HepG2細胞+DOX+GSH。MTT 法檢測 HepG2細胞生長,流式細胞儀檢測細胞早期凋亡率,實時熒光定量 PCR 法檢測 NF-κB p65 mRNA 的錶達,Western 印跡檢測 NF-κB p65及 Bcl-2的錶達。結果(1)DOX 組和 DOX+GSH 組作用細胞24 h、48 h、72 h 均能顯著抑製 HepG2細胞增殖,DOX 組抑製率顯著高于 DOX+GSH 組(P <0.05),且抑製率具有時間和劑量依賴性。(2)對照組凋亡率為(0.733±0.153)%,DOX 組凋亡率為(28.400±0.007)%,DOX+GSH 組凋亡率為(15.500±0.006)%;DOX 組、DOX+GSH 組分彆與對照組相比,差異有統計學意義(P <0.01);DOX 組與 DOX+GSH 組相比,差異有統計學意義(P <0.01)。(3)兩組在處理細胞24 h 後,DOX+GSH 組 NF-κB p65mRNA 錶達顯著高于 DOX組(P <0.05)。(4)DOX 組 NF-κB p65、Bcl-2錶達較對照組增高,DOX+GSH 組錶達較 DOX 組錶達增高。結論 GSH 與DOX 聯閤使用可導緻 DOX 化療效果下降,其機製可能是通過進一步上調 NF-κB p65和 Bcl-2的錶達實現的。臨床上使用DOX 化療的腫瘤患者應避免同時使用 GSH。
목적:연구환원형곡광감태(GSH)대다유비성(DOX)처리 HepG2세포주증식급조망적영향;탐색핵전록인자(NF)-κB p65、Bcl-2재 DOX 단약급연합 GSH 유도 HepG2세포조망후적표체급기의의。방법실험분4조,공백조:배양액,대조조:배양액+HepG2세포+RPMl l640배양기,DOX 조:배양액+HepG2세포+DOX,DOX+GSH 조:배양액+HepG2세포+DOX+GSH。MTT 법검측 HepG2세포생장,류식세포의검측세포조기조망솔,실시형광정량 PCR 법검측 NF-κB p65 mRNA 적표체,Western 인적검측 NF-κB p65급 Bcl-2적표체。결과(1)DOX 조화 DOX+GSH 조작용세포24 h、48 h、72 h 균능현저억제 HepG2세포증식,DOX 조억제솔현저고우 DOX+GSH 조(P <0.05),차억제솔구유시간화제량의뢰성。(2)대조조조망솔위(0.733±0.153)%,DOX 조조망솔위(28.400±0.007)%,DOX+GSH 조조망솔위(15.500±0.006)%;DOX 조、DOX+GSH 조분별여대조조상비,차이유통계학의의(P <0.01);DOX 조여 DOX+GSH 조상비,차이유통계학의의(P <0.01)。(3)량조재처리세포24 h 후,DOX+GSH 조 NF-κB p65mRNA 표체현저고우 DOX조(P <0.05)。(4)DOX 조 NF-κB p65、Bcl-2표체교대조조증고,DOX+GSH 조표체교 DOX 조표체증고。결론 GSH 여DOX 연합사용가도치 DOX 화료효과하강,기궤제가능시통과진일보상조 NF-κB p65화 Bcl-2적표체실현적。림상상사용DOX 화료적종류환자응피면동시사용 GSH。
Objective To study the effects of reduced glutathione (GSH)on doxorubicin (DOX)induced proliferation and apoptosis in HepG2,and to investigate the expression of NF-κB p65,Bcl-2 in apoptostic HepG2 induced by DOX monotherapy and DOX combination with GSH,respectively.Methods Four groups were devided,including blank group:RPMl l640 medium only;control group:HepG2 cells were cultured in the same volume of medium;DOX group:cells were cultured in medium containing DOX;DOX+GSH group:cells were cultured in medium containing DOX and GSH.MTT assay was performed to probe proliferation of HepG2 cell.Flow cytometry was carried out to determine early apoptosis rate of HepG2 cells.Real-time polymerase chain reaction was administered to investigate NF-κB p65 mRNA expression. Western blotting was used to document protein expression of NF-κB p65 and Bcl-2.The experimental data was analyzed by SPSS version 19.0 software.Results (1)HepG2 cell proliferation was inhibited time-and dose-dependently in both DOX group and DOX + GSH group at 24 h or 48 h or 72 h.Additionally,the proliferation rate was higher in DOX group than that in DOX+GSH group(P <0.05).(2)The apoptosis rate of HepG2 was (0.733±0.153)% in control group,(28.400 ±0.007)% in DOX group,(15.500±0.006)% in DOX+GSH group,respectively,which showed statistically significant difference between the later two groups and control group (P <0.01 ),as well as between DOX group and DOX+ GSH group (P <0.01).(3)The mRNA expression of NF-κB p65 in DOX+GSH group was higher than that in DOX group (P <0.05).(4)The protein expression of NF-κB p65 and Bcl-2 in DOX+GSH group was higher than that in DOX group (P <0.05 ).Conclusion DOX and GSH combination therapy could attenuate chemotherapeutic effect though a potential mechanism by increasing expression of NF-κB p65 and Bcl-2.The combination of DOX and GSH in chemotherapy for cancer patients should be avoided.