中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2014年
4期
424-428
,共5页
蒲华清%王秉翔%杜爱玲%李英杰%张志勉
蒲華清%王秉翔%杜愛玲%李英傑%張誌勉
포화청%왕병상%두애령%리영걸%장지면
癌,肝细胞%抗肿瘤联合化疗方案%细胞凋亡%细胞低氧
癌,肝細胞%抗腫瘤聯閤化療方案%細胞凋亡%細胞低氧
암,간세포%항종류연합화료방안%세포조망%세포저양
Carcinoma,hepatocellular%Antineoplastic combined chemotherapy protocols%Apoptosis%Cell hypoxia
目的 对比6-姜酚在正常和低氧、低糖模式下对于人肝癌细胞株(HepG-2)细胞的杀伤和化疗增敏作用. 方法 通过浓度为5、10、15、20、40、60 mg/L阿霉素(多柔比星,ADM)和25、50、100、200 μmol/L 6-姜酚刺激培养至对数期的HepG-2细胞,采用CCK-8试剂盒检测HepG-2细胞增殖反应.通过流式细胞术结合细胞凋亡检测试剂盒双标染色法检测药物作用后细胞的凋亡情况.采用实时聚合酶链式反应(real-time PCR)检测bel-2、bax及birc-5 mRNA的表达情况. 结果 6-姜酚、ADM作用于HepG-2细胞后,细胞生长受到抑制.两种模式下6-姜酚组、阿霉素组的抑制率随浓度的升高而增强,抑制率具有浓度依赖性.正常培养条件下对照组、6-姜酚组、ADM组及6-姜酚+ADM组HepG-2细胞凋亡百分比分别为(7.98±0.76)％、(9.63±1.00)％、(12.70±2.13)％、(19.92±1.41)％;低氧低糖条件下对照组、6-姜酚组、ADM组及6-姜酚+ADM组HepG-2细胞凋亡百分比分别为(13.92±2.02)％、(19.36±1.22)％、(27.87±0.99)％、(38.63±2.25)％,两种培养条件下各浓度组细胞凋亡百分比均较对照组增高,差异有统计学意义(t值分别为7.250、5.259、12.185、8.140、15.000、47.576,P值分别为0.019、0.034、0.007、0.015、0.004、0.000),两药联用组的凋亡数最高,且低氧、低糖各浓度组的凋亡数高于正常培养各浓度组.real-time PCR检测结果表明,正常培养条件下与对照组比较,实验组bcl-2、birc-5表达降低,bax表达无变化.低氧低糖条件下与对照组比较实验组bcl-2、birc-5表达降低,bax表达增高.在低氧低糖环境下bcl-2的表达较正常培养条件下的表达增高,而bax、birc-5表达减少,bcl-2/bax高于正常培养组. 结论 6-姜酚可能通过下调birc-5的表达,降低Survivin蛋白抑制肿瘤细胞的凋亡的能力对HepG-2细胞产生杀伤和化疗增敏作用,在低氧低糖环境中这种作用表现地更为明显.
目的 對比6-薑酚在正常和低氧、低糖模式下對于人肝癌細胞株(HepG-2)細胞的殺傷和化療增敏作用. 方法 通過濃度為5、10、15、20、40、60 mg/L阿黴素(多柔比星,ADM)和25、50、100、200 μmol/L 6-薑酚刺激培養至對數期的HepG-2細胞,採用CCK-8試劑盒檢測HepG-2細胞增殖反應.通過流式細胞術結閤細胞凋亡檢測試劑盒雙標染色法檢測藥物作用後細胞的凋亡情況.採用實時聚閤酶鏈式反應(real-time PCR)檢測bel-2、bax及birc-5 mRNA的錶達情況. 結果 6-薑酚、ADM作用于HepG-2細胞後,細胞生長受到抑製.兩種模式下6-薑酚組、阿黴素組的抑製率隨濃度的升高而增彊,抑製率具有濃度依賴性.正常培養條件下對照組、6-薑酚組、ADM組及6-薑酚+ADM組HepG-2細胞凋亡百分比分彆為(7.98±0.76)％、(9.63±1.00)％、(12.70±2.13)％、(19.92±1.41)％;低氧低糖條件下對照組、6-薑酚組、ADM組及6-薑酚+ADM組HepG-2細胞凋亡百分比分彆為(13.92±2.02)％、(19.36±1.22)％、(27.87±0.99)％、(38.63±2.25)％,兩種培養條件下各濃度組細胞凋亡百分比均較對照組增高,差異有統計學意義(t值分彆為7.250、5.259、12.185、8.140、15.000、47.576,P值分彆為0.019、0.034、0.007、0.015、0.004、0.000),兩藥聯用組的凋亡數最高,且低氧、低糖各濃度組的凋亡數高于正常培養各濃度組.real-time PCR檢測結果錶明,正常培養條件下與對照組比較,實驗組bcl-2、birc-5錶達降低,bax錶達無變化.低氧低糖條件下與對照組比較實驗組bcl-2、birc-5錶達降低,bax錶達增高.在低氧低糖環境下bcl-2的錶達較正常培養條件下的錶達增高,而bax、birc-5錶達減少,bcl-2/bax高于正常培養組. 結論 6-薑酚可能通過下調birc-5的錶達,降低Survivin蛋白抑製腫瘤細胞的凋亡的能力對HepG-2細胞產生殺傷和化療增敏作用,在低氧低糖環境中這種作用錶現地更為明顯.
목적 대비6-강분재정상화저양、저당모식하대우인간암세포주(HepG-2)세포적살상화화료증민작용. 방법 통과농도위5、10、15、20、40、60 mg/L아매소(다유비성,ADM)화25、50、100、200 μmol/L 6-강분자격배양지대수기적HepG-2세포,채용CCK-8시제합검측HepG-2세포증식반응.통과류식세포술결합세포조망검측시제합쌍표염색법검측약물작용후세포적조망정황.채용실시취합매련식반응(real-time PCR)검측bel-2、bax급birc-5 mRNA적표체정황. 결과 6-강분、ADM작용우HepG-2세포후,세포생장수도억제.량충모식하6-강분조、아매소조적억제솔수농도적승고이증강,억제솔구유농도의뢰성.정상배양조건하대조조、6-강분조、ADM조급6-강분+ADM조HepG-2세포조망백분비분별위(7.98±0.76)％、(9.63±1.00)％、(12.70±2.13)％、(19.92±1.41)％;저양저당조건하대조조、6-강분조、ADM조급6-강분+ADM조HepG-2세포조망백분비분별위(13.92±2.02)％、(19.36±1.22)％、(27.87±0.99)％、(38.63±2.25)％,량충배양조건하각농도조세포조망백분비균교대조조증고,차이유통계학의의(t치분별위7.250、5.259、12.185、8.140、15.000、47.576,P치분별위0.019、0.034、0.007、0.015、0.004、0.000),량약련용조적조망수최고,차저양、저당각농도조적조망수고우정상배양각농도조.real-time PCR검측결과표명,정상배양조건하여대조조비교,실험조bcl-2、birc-5표체강저,bax표체무변화.저양저당조건하여대조조비교실험조bcl-2、birc-5표체강저,bax표체증고.재저양저당배경하bcl-2적표체교정상배양조건하적표체증고,이bax、birc-5표체감소,bcl-2/bax고우정상배양조. 결론 6-강분가능통과하조birc-5적표체,강저Survivin단백억제종류세포적조망적능력대HepG-2세포산생살상화화료증민작용,재저양저당배경중저충작용표현지경위명현.
Objective To compare the cell-killing and sensitization effect of 6-gingerol on human hepatoma carcinoma (HepG-2) cell in normal mode versus hypoxia-hypoglycemia mode in chemotherapy.Methods The HepG-2 cells was cultured to logarithmic phase and treated with adriamyein doxorubicin hydrochloride (ADM) (5,10,15,20,40,60 mg/L) and 6-gingerol(25,50,100,200 μmol/L)in different concentrations.Then the cell counting kit (CCK-8) assay kit was used to determine the proliferation inhibition of HepG-2 cells.Cell apoptosis was detected by combining flow cytometry and AnnexinV-FITC PI double staining after treated with different drugs.The expressions of bcl-2,bax and birc-5 mRNA in HepG-2 cells was detected by real time fluorescence quantitative polymerase chain reaction(RT-PCR)assay.Results 6-gingerol and ADM had a certain degree of growth inhibition on HepG-2 cells.In two modes,the inhibition ratios of the 6-gingerol and ADM were both increased along with the increase of the concentration,which showed a dose-dependent manner.Flow cytometry analysis demonstrated that the apoptosis rate in the control group,6-gingerol group,ADM group and the 6-gingerol+ADM group in the normal mode was (7.98±0.76)％,(9.63 ± 1.00) ％,(12.70 ± 2.13) ％ and (19.92 ± 1.41) ％ respectively.The apoptosis rate in the control group,6-gingerol group,ADM group and the 6-gingerol+ ADM group in the hypoxia-hypoglycemia mode was (13.92 ± 2.02)％,(19.36 ±-1.22)％,(27.87 ± 0.99)％ and (38.63 ± 2.25)％ respectively.It demonstrated that the apoptosis rate was increased in the experimental groups as compared to the control group under the two culture conditions(the normal mode and the hypoxiahypoglycemia mode)(t=7.250,5.259,12.185,8.140,15.000,47.576,respectively,all P＜0.05,0.01 or 0.001).The combination group had the highest number of apoptosis cells,and the number of apoptosis cells was higher in hypoxia-hypoglycemia group than in normal culture group.Real-time PCR analysis showed that,compared with the control group,the expressions of bcl-2 and birc-5 mRNA were decreased and the expression of bax mRNA had no significant changes in experimental group under the normal culture conditions.The expressions of bcl-2 and birc-5 mRNA were significantly decreased and the expression of bax mRNA was increased in experimental group as compared with the control group under the hypoxia-hypoglycemia conditions.Under the hypoxiahypoglycemia environment,the expression of bcl-2 mRNA was significantly increased,the expressions of bax and birc-5 mRNA was significantly reduced,and the ratio of bcl-2 and bax was significantly increased as compared with the normal culture conditions.Conclusions 6 gingerol may decrease the inhibitory effect of survivin protein on tumor cells apoptosis by reduced the expression of birc-5,which generates the cell-killing and sensitizing effect on HepG-2 cell in chemotherapy.This performance is more obvious in the hypoxia-hypoglycemia environment.
