中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2010年
10期
770-776
,共7页
郑建伟%薛新波%王从俊%张晓梅%陈堃%李雁%于愿%肖朝文%彭志海%易继林%吴在德
鄭建偉%薛新波%王從俊%張曉梅%陳堃%李雁%于願%肖朝文%彭誌海%易繼林%吳在德
정건위%설신파%왕종준%장효매%진곤%리안%우원%초조문%팽지해%역계림%오재덕
肝细胞癌%阿霉素%多药耐药%凋亡
肝細胞癌%阿黴素%多藥耐藥%凋亡
간세포암%아매소%다약내약%조망
Hepatocellular carcinoma%Adriamycin%Multidrug resistance%Apoptosis
目的 探讨黑色素瘤分化相关基因-7/白介素-24(MDA-7/IL-24)基因联合阿霉素杀伤肝癌细胞HepG2,并逆转HepG2多药耐药的机制.方法 以人肝癌细胞系HepG2为实验对象,使用MTT法和流式细胞仪比较Ad.mda-7联合阿霉素处理组与阿霉素组、Ad.mda-7组对肝癌细胞HepG2的作用差异.阐明MDA-7/IL-24对多药耐药的逆转作用的机制.实时定量PCR检测MDR-1、STAT-3、BCL-2、BAXmRNA的变化.Western blot检测gp-170、stat3、P-stat-3、PKB、bcl-2、hax蛋白的表达的变化.结果 低浓度的Ad.mda-7联合正常肝细胞半抑制浓度浓度的阿霉素(1.5 mg/L)使得细胞抑制率从阿霉素组的17.46%上升到79.5%,生长抑制逆转4.55倍(P＜0.05).联合化疗组MDR-1mRNA相对表达量从16.49±0.11下降至5.48±0.05.STAT-3 mRNA相对表达量从13.17±0.08上升至21.57±0.11.BCL-2及BAX表达与其他实验组相比较差异均有显著统计学意义(P＜0.05).联合实验组P-170蛋白的表达量较其他组明显降低,PKB蛋白表达量明显降低,磷酸化stat-3蛋白的表达量增加.结论 Ad.mda-7具有逆转肝癌细胞HepG2多药耐药的作用.其在下调MDR-1mRNA表达的同时,通过抑制PKB蛋白和活化STAT-3信号通路的表达降低促进肝癌细胞凋亡.
目的 探討黑色素瘤分化相關基因-7/白介素-24(MDA-7/IL-24)基因聯閤阿黴素殺傷肝癌細胞HepG2,併逆轉HepG2多藥耐藥的機製.方法 以人肝癌細胞繫HepG2為實驗對象,使用MTT法和流式細胞儀比較Ad.mda-7聯閤阿黴素處理組與阿黴素組、Ad.mda-7組對肝癌細胞HepG2的作用差異.闡明MDA-7/IL-24對多藥耐藥的逆轉作用的機製.實時定量PCR檢測MDR-1、STAT-3、BCL-2、BAXmRNA的變化.Western blot檢測gp-170、stat3、P-stat-3、PKB、bcl-2、hax蛋白的錶達的變化.結果 低濃度的Ad.mda-7聯閤正常肝細胞半抑製濃度濃度的阿黴素(1.5 mg/L)使得細胞抑製率從阿黴素組的17.46%上升到79.5%,生長抑製逆轉4.55倍(P＜0.05).聯閤化療組MDR-1mRNA相對錶達量從16.49±0.11下降至5.48±0.05.STAT-3 mRNA相對錶達量從13.17±0.08上升至21.57±0.11.BCL-2及BAX錶達與其他實驗組相比較差異均有顯著統計學意義(P＜0.05).聯閤實驗組P-170蛋白的錶達量較其他組明顯降低,PKB蛋白錶達量明顯降低,燐痠化stat-3蛋白的錶達量增加.結論 Ad.mda-7具有逆轉肝癌細胞HepG2多藥耐藥的作用.其在下調MDR-1mRNA錶達的同時,通過抑製PKB蛋白和活化STAT-3信號通路的錶達降低促進肝癌細胞凋亡.
목적 탐토흑색소류분화상관기인-7/백개소-24(MDA-7/IL-24)기인연합아매소살상간암세포HepG2,병역전HepG2다약내약적궤제.방법 이인간암세포계HepG2위실험대상,사용MTT법화류식세포의비교Ad.mda-7연합아매소처리조여아매소조、Ad.mda-7조대간암세포HepG2적작용차이.천명MDA-7/IL-24대다약내약적역전작용적궤제.실시정량PCR검측MDR-1、STAT-3、BCL-2、BAXmRNA적변화.Western blot검측gp-170、stat3、P-stat-3、PKB、bcl-2、hax단백적표체적변화.결과 저농도적Ad.mda-7연합정상간세포반억제농도농도적아매소(1.5 mg/L)사득세포억제솔종아매소조적17.46%상승도79.5%,생장억제역전4.55배(P＜0.05).연합화료조MDR-1mRNA상대표체량종16.49±0.11하강지5.48±0.05.STAT-3 mRNA상대표체량종13.17±0.08상승지21.57±0.11.BCL-2급BAX표체여기타실험조상비교차이균유현저통계학의의(P＜0.05).연합실험조P-170단백적표체량교기타조명현강저,PKB단백표체량명현강저,린산화stat-3단백적표체량증가.결론 Ad.mda-7구유역전간암세포HepG2다약내약적작용.기재하조MDR-1mRNA표체적동시,통과억제PKB단백화활화STAT-3신호통로적표체강저촉진간암세포조망.
Objective To explore the mechanism of melanoma differentiation associated gene-7(mda-7) in combination with adriamycin(ADM) killing the HCC HepG2 cells and reversing their multidrug resistance (MDR). Methods The experiment was conducted in three groups including the combined group, ADM group and mda-7 group. MTT assay and FCM were used to determine the differences among the 3 groups and clarify the reversing effect of combined treatment on multidrug resistance of the tumor cells. Expression levels of MDR-1, STAT-3, BCL-2, BAXmRNA were determined with real-time PCR. Western blotting was performed to observe the changes of proteins gp-l70, stat3,P-stat3, PKB, bcl-2,bax in all 3 groups. Result After transfection with 100VP/cell Ad. mda-7,the growth suppression rate of HepG2 treated by ADM (1.5 mg/L) rose from 17.46% to 79. 5%.According to the changes, killed HepG2 cells were increased by a factor of 4.55. times. MDR-1 mRNA was decreased from (16.49 ± 0. 11) to (5.48±0.05) and STAT-3 mRNA increased from (13.17±0. 08) to (21. 57±0. 11)(P＜0.05). Western blotting also showed that P-170 and PKB was decreased and the phosphorylation-stat-3 increased after the combined treatment. Conclusion Ad.mda-7 can reverse the multidrug resistance HepG2 cells. It inhibits the expression of MDR-1 mRNA,then arrests PKB protein and the signaling pathway of active stat-3 to induce apoptosis of HCC cells.
