中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2014年
5期
307-311
,共5页
胰腺肿瘤%浅蓝菌素%吉西他滨%药物协同作用%细胞凋亡
胰腺腫瘤%淺藍菌素%吉西他濱%藥物協同作用%細胞凋亡
이선종류%천람균소%길서타빈%약물협동작용%세포조망
Pancreatic neoplasms%Cerulenin%Gemcitabine%Drug synergism%Apoptosis
目的 观察浅蓝菌素联合吉西他滨对胰腺癌细胞BxPC3生长抑制的协同作用,探讨其机制.方法 以10μg/ml浅蓝菌素、20μmol/L吉西他滨、10 μg/ml浅蓝菌素+20μg/L吉西他滨分别处理对数生长期的人胰腺癌BxPC3细胞48 h,以不经药物处理的细胞作为对照.应用CCK-8法检测各组细胞增殖,AnnexinV/PI双染法检测细胞早期凋亡率,RT-PCR法检测Bcl-2、Bax mRNA表达,蛋白质印迹法检测Bcl-2、Bax蛋白表达.结果 对照组、浅蓝菌素组、吉西他滨组、联合处理组48 h的生长抑制率分别为0、(51.28±1.84)％、(53.59±1.62)％、(84.57 ±1.01)％;细胞早期凋亡率为(0.83±0.31)％、(31.37±1.04)％、(38.33±0.75)％、(69.43±0.83)％;BxPC3细胞Bcl-2 mRNA表达量分别为0.67±0.01、0.44±0.01、0.36±0.08、0.27 ±0.07;Bax mRNA表达量为0.14±0.01、0.31±0.02、0.32±0.03、0.91±0.06;Bcl-2 mRNA/Bax mRNA比值为4.78±0.13、1.39±0.04、1.15±0.02、0.30±0.02;Bcl-2蛋白表达量分别为1.24±0.04、0.51±0.02、0.42±0.02、0.13±0.01;Bax蛋白表达量为0.20±0.05、0.47±0.01、0.54±0.01、1.21±0.03;Bcl-2/Bax比值为6.00±0.11、1.11±0.01、0.77±0.03、0.10±0.06.各处理组与对照组,联合处理组与单药处理组的差异均有统计学意义(P值均＜0.01).结论 浅蓝菌素与吉西他滨对人胰腺癌BxPC3细胞的生长抑制具有协同作用,其机制可能通过上调Bax基因表达,下调Bcl-2基因表达,降低Bcl-2/Bax比值而促进细胞凋亡所致.
目的 觀察淺藍菌素聯閤吉西他濱對胰腺癌細胞BxPC3生長抑製的協同作用,探討其機製.方法 以10μg/ml淺藍菌素、20μmol/L吉西他濱、10 μg/ml淺藍菌素+20μg/L吉西他濱分彆處理對數生長期的人胰腺癌BxPC3細胞48 h,以不經藥物處理的細胞作為對照.應用CCK-8法檢測各組細胞增殖,AnnexinV/PI雙染法檢測細胞早期凋亡率,RT-PCR法檢測Bcl-2、Bax mRNA錶達,蛋白質印跡法檢測Bcl-2、Bax蛋白錶達.結果 對照組、淺藍菌素組、吉西他濱組、聯閤處理組48 h的生長抑製率分彆為0、(51.28±1.84)％、(53.59±1.62)％、(84.57 ±1.01)％;細胞早期凋亡率為(0.83±0.31)％、(31.37±1.04)％、(38.33±0.75)％、(69.43±0.83)％;BxPC3細胞Bcl-2 mRNA錶達量分彆為0.67±0.01、0.44±0.01、0.36±0.08、0.27 ±0.07;Bax mRNA錶達量為0.14±0.01、0.31±0.02、0.32±0.03、0.91±0.06;Bcl-2 mRNA/Bax mRNA比值為4.78±0.13、1.39±0.04、1.15±0.02、0.30±0.02;Bcl-2蛋白錶達量分彆為1.24±0.04、0.51±0.02、0.42±0.02、0.13±0.01;Bax蛋白錶達量為0.20±0.05、0.47±0.01、0.54±0.01、1.21±0.03;Bcl-2/Bax比值為6.00±0.11、1.11±0.01、0.77±0.03、0.10±0.06.各處理組與對照組,聯閤處理組與單藥處理組的差異均有統計學意義(P值均＜0.01).結論 淺藍菌素與吉西他濱對人胰腺癌BxPC3細胞的生長抑製具有協同作用,其機製可能通過上調Bax基因錶達,下調Bcl-2基因錶達,降低Bcl-2/Bax比值而促進細胞凋亡所緻.
목적 관찰천람균소연합길서타빈대이선암세포BxPC3생장억제적협동작용,탐토기궤제.방법 이10μg/ml천람균소、20μmol/L길서타빈、10 μg/ml천람균소+20μg/L길서타빈분별처리대수생장기적인이선암BxPC3세포48 h,이불경약물처리적세포작위대조.응용CCK-8법검측각조세포증식,AnnexinV/PI쌍염법검측세포조기조망솔,RT-PCR법검측Bcl-2、Bax mRNA표체,단백질인적법검측Bcl-2、Bax단백표체.결과 대조조、천람균소조、길서타빈조、연합처리조48 h적생장억제솔분별위0、(51.28±1.84)％、(53.59±1.62)％、(84.57 ±1.01)％;세포조기조망솔위(0.83±0.31)％、(31.37±1.04)％、(38.33±0.75)％、(69.43±0.83)％;BxPC3세포Bcl-2 mRNA표체량분별위0.67±0.01、0.44±0.01、0.36±0.08、0.27 ±0.07;Bax mRNA표체량위0.14±0.01、0.31±0.02、0.32±0.03、0.91±0.06;Bcl-2 mRNA/Bax mRNA비치위4.78±0.13、1.39±0.04、1.15±0.02、0.30±0.02;Bcl-2단백표체량분별위1.24±0.04、0.51±0.02、0.42±0.02、0.13±0.01;Bax단백표체량위0.20±0.05、0.47±0.01、0.54±0.01、1.21±0.03;Bcl-2/Bax비치위6.00±0.11、1.11±0.01、0.77±0.03、0.10±0.06.각처리조여대조조,연합처리조여단약처리조적차이균유통계학의의(P치균＜0.01).결론 천람균소여길서타빈대인이선암BxPC3세포적생장억제구유협동작용,기궤제가능통과상조Bax기인표체,하조Bcl-2기인표체,강저Bcl-2/Bax비치이촉진세포조망소치.
Objective To investigate the effect and mechanism of action in pancreatic cancer BxPC3 cell treated by cerulenin in combination with gemcitabine.Methods BxPC3 cells were cultivated with 10 μg/ml cerulenin,20 μmol/L gemcitabine or 10 μg/ml cerulenin + 20 μmol/L gemcitabine for 48 h,and cells without treatment were control.Cell proliferation was detected by CCK-8 assay,and early apoptosis was detected by AnnexinV/PI double staining method.The expression of Bcl-2 mRNA and Bax mRNA were detected by RT-PCR,and the protein level of Bcl-2,Bax were detected by Western Blot.Results The inhibition rate of BxPC3 cells were 0,(51.28 ± 1.84) ％,(53.59 ± 1.62) ％,(84.57 ± 1.01) ％ in control,cerulenin group,gemcitabine group,combination group; and the rate of early apoptosis was (0.83 ± 0.31) ％,(31.37 ± 1.04) ％,(38.33 ± 0.75) ％,(69.43 ± 0.83) ％,and the expression of Bcl-2 mRNA was 0.67 ± 0.01,0.44 ±0.01,0.36 ±0.08,0.27 ±0.07,and the expression of Bax mRNA was 0.14 ±0.01,0.31 ± 0.02,0.32 ± 0.03,0.91 ± 0.06 ; while the ratio of Bcl-2 mRNA/Bax mRNA was 4.78 ± 0.13,1.39 ± 0.04,1.15 ± 0.02,0.30 ± 0.02 ; the expression of Bcl-2 protein was 1.24 ± 0.04,0.51 ± 0.02,0.42 ± 0.02,0.13 ±0.01 ; and the expression of Bax protein was 0.20 ± 0.05,0.47 ± 0.01,0.54 ± 0.01,1.21 ± 0.03 ; while the ratio of Bcl-2/Bax was 6.00 ± 0.11,1.11 ± 0.01,0.77 ± 0.03,0.10 ± 0.06.And the difference among the groups was statistically significant (P ＜ 0.01).Conclusions Cerulenin combined with gemcitabin has synergistic effect on the inhibition of BxPC cells proliferation.Its mechanisms may be up-regulation of Bax,down-regulation of Bcl-2,and promoting apoptosis of cells through the decrease of Bcl-2/Bax ratio.
