INTRODUCTIONThe treatment of human epithelial malignancies islimited by drug resistance and toxic and side effects,which results in the failure in the treatment ofmajority of advanced cancer victims.To seek for anew,a...INTRODUCTIONThe treatment of human epithelial malignancies islimited by drug resistance and toxic and side effects,which results in the failure in the treatment ofmajority of advanced cancer victims.To seek for anew,and specific antineoplastic therapy willprovide hope for tumor treatment.Althoughdisordered intermediary metabolism in cancer cellshas been known for many years,much of the展开更多
目的:探讨脂肪酸合成酶(FAS)的抑制剂浅蓝菌素对前列腺癌细胞株PC-3的生长抑制作用。方法:采用人前列腺癌细胞株PC-3在体外实验中用MTT法观察浅蓝菌素对细胞株的生长抑制作用;流式细胞术(FCM)对细胞株的凋亡及细胞周期的变化进行检测;...目的:探讨脂肪酸合成酶(FAS)的抑制剂浅蓝菌素对前列腺癌细胞株PC-3的生长抑制作用。方法:采用人前列腺癌细胞株PC-3在体外实验中用MTT法观察浅蓝菌素对细胞株的生长抑制作用;流式细胞术(FCM)对细胞株的凋亡及细胞周期的变化进行检测;蛋白免疫印迹(W estern b lotting)法检测浅蓝菌素对PC-3细胞中FAS蛋白水平表达的影响。结果:PC-3细胞在浅蓝菌素的作用下,细胞增殖被明显阻遏,并呈现剂量效应关系,在浓度为2.5mg/L、5.0mg/L、10.0mg/L、20.0mg/L的浅蓝菌素作用下,PC-3细胞24h的抑制率分别为(34.78±2.47)%、(46.76±3.18)%、(58.13±3.55)%、(65.31±2.81)%,与对照组比较有统计学意义(P<0.05)。流式细胞仪显示,细胞经浅蓝菌素作用后,细胞凋亡增多,PC-3细胞在浅蓝菌素浓度5.0mg/L三个时间组(24h,48h,72h)的细胞凋亡率分别为(28.29±1.88)%、(44.73±2.69)%、(61.25±1.43)%;且G0/G1期细胞比例增加,S期和G2/M期细胞比例减少。蛋白免疫印迹实验中,随着浅蓝菌素作用时间的延长,PC-3细胞中的FAS蛋白水平表达量明显减少,药物作用时间与细胞中蛋白水平表达量呈负相关。结论:浅蓝菌素能抑制PC-3细胞生长,且其呈现时间和浓度依赖性。展开更多
Objective： To determine fatty acid synthase （FAS） expression in human multiple myeloma and verify its potential as a therapeutic target in multiple myeloma. Methods： FAS expression was determined by immunohistoche...Objective： To determine fatty acid synthase （FAS） expression in human multiple myeloma and verify its potential as a therapeutic target in multiple myeloma. Methods： FAS expression was determined by immunohistochemistry, reverse-transcription polymerase chain reaction （RT-PCR） and immunoblot analysis in bone marrow samples obtained from 27 patients with multiple myeloma （MM patients） and peripheral blood mononuclear cells （PBMCs） obtained from 12 healthy donors In parallel, additional analyses were performed on 2 human multiple myeloma cell lines, U266 and RPM18226. U266 cells were treated with cerulenin at various concentrations （5 to 320 μg/ml） for 24 h, and metabolic activity was measured by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assays. Apoptosis was evaluated by dual Annexin V/Pl （propidium iodide） labeling and flow cytometry （FCM） in U266 cells treated with 20 μg/ml cerulenin for 12 h or 24 h. Results： By immunohistochemistry, we found that 19 of 27 bone marrow samples obtained from MM patients expressed significantly high levels of FAS. Similarly, by RT-PCR, 22 of 27 bone marrow samples obtained from MM patients, U266 and RPM18226 showed FAS expression, whereas PBMC samples from 12 healthy donors did not express detectable level of FAS. FAS protein expression was confirmed by immunoblot analysis in 16 of 27 bone marrow samples obtained from MM patients, U266 and RPM18226 cell lines, and no FAS protein expression was detected in PBMC samples from 12 healthy donors. U266 cells were highly sensitive to cerulenin treatment, with a dosage-related effect on metabolic activity, as a measure for cell proliferation. U266 cells treated with 20 μg/ml cerulenin for 12 and 24 h also showed early sign of apoptosis with 56.9% and 69.3% Annexin V^＋/Pl cells, and late apoptotic and necrotic cells with 3.2% and 17.6% Annexin V^＋/Pl^＋ cells. Conclusion： Increased FAS expression existed in multiple myeloma samples and human myeloma cell lines. Cerulenin greatly inhibited metabolic activity/cell proliferation of U266 cells and induced apoptosis, suggesting that FAS is an effective target for pharmacological therapy in human multiple myeloma.展开更多
目的研究浅蓝菌素(cerulenin)对人成骨肉瘤细胞U2-OS在裸鼠体内生长抑制作用。方法建立人成骨肉瘤细胞U2-OS BALB/C裸鼠瘤模型。18只成瘤裸鼠按随机数字表分成3组,每组6只;对照组(DMSO溶剂)、低剂量cerulenin组(0.2 mL cerulenin配制液4...目的研究浅蓝菌素(cerulenin)对人成骨肉瘤细胞U2-OS在裸鼠体内生长抑制作用。方法建立人成骨肉瘤细胞U2-OS BALB/C裸鼠瘤模型。18只成瘤裸鼠按随机数字表分成3组,每组6只;对照组(DMSO溶剂)、低剂量cerulenin组(0.2 mL cerulenin配制液40 mg.kg-1.次-1)、高剂量cerulenin组(0.2 mL cerulenin配制液80 mg.kg-1.次-1)于腹腔隔日注射连续6次。动态观察3组肿瘤体积及裸鼠质量变化,用药结束后18 d处死裸鼠,测量瘤重、计算瘤重抑制率,测量肿瘤体积、计算肿瘤体积抑制率。肿瘤组织行形态学观察。TUNEL法检测肿瘤组织细胞凋亡情况。结果高、低剂量cerulenin组肿瘤体积抑制率分别为51.2%和24.2%,瘤重抑制率分别为49.1%和25.5%。各组间瘤体体积与瘤重比较,差异均有统计学意义(P<0.05)。瘤体组织学显示cerulenin组较对照组出现明显的细胞凋亡的超微结构改变,瘤细胞体积较小,多数细胞微绒毛消失,胞质密度增高。TUNEL法显示高、低剂量cerulenin组,对照组癌细胞凋亡率为(17.1±1.2)%、(10.3±0.8)%、(3.5±0.5)%,3组比较差异均有统计学意义(P<0.05)。结论 cerulenin能够有效诱导人成骨肉瘤细胞U2-OS在裸鼠体内凋亡,抑制肿瘤的生长。展开更多
基金the Applied Science Foundation of Jiangsu Province,China,No.BJ97071
文摘INTRODUCTIONThe treatment of human epithelial malignancies islimited by drug resistance and toxic and side effects,which results in the failure in the treatment ofmajority of advanced cancer victims.To seek for anew,and specific antineoplastic therapy willprovide hope for tumor treatment.Althoughdisordered intermediary metabolism in cancer cellshas been known for many years,much of the
文摘目的:探讨脂肪酸合成酶(FAS)的抑制剂浅蓝菌素对前列腺癌细胞株PC-3的生长抑制作用。方法:采用人前列腺癌细胞株PC-3在体外实验中用MTT法观察浅蓝菌素对细胞株的生长抑制作用;流式细胞术(FCM)对细胞株的凋亡及细胞周期的变化进行检测;蛋白免疫印迹(W estern b lotting)法检测浅蓝菌素对PC-3细胞中FAS蛋白水平表达的影响。结果:PC-3细胞在浅蓝菌素的作用下,细胞增殖被明显阻遏,并呈现剂量效应关系,在浓度为2.5mg/L、5.0mg/L、10.0mg/L、20.0mg/L的浅蓝菌素作用下,PC-3细胞24h的抑制率分别为(34.78±2.47)%、(46.76±3.18)%、(58.13±3.55)%、(65.31±2.81)%,与对照组比较有统计学意义(P<0.05)。流式细胞仪显示,细胞经浅蓝菌素作用后,细胞凋亡增多,PC-3细胞在浅蓝菌素浓度5.0mg/L三个时间组(24h,48h,72h)的细胞凋亡率分别为(28.29±1.88)%、(44.73±2.69)%、(61.25±1.43)%;且G0/G1期细胞比例增加,S期和G2/M期细胞比例减少。蛋白免疫印迹实验中,随着浅蓝菌素作用时间的延长,PC-3细胞中的FAS蛋白水平表达量明显减少,药物作用时间与细胞中蛋白水平表达量呈负相关。结论:浅蓝菌素能抑制PC-3细胞生长,且其呈现时间和浓度依赖性。
基金Project supported by the Medicine and Health Research Fund of Zhejiang Province(No.2007B091)the Office of Education of Zhejiang Province,China(No.20070104)
文摘Objective： To determine fatty acid synthase （FAS） expression in human multiple myeloma and verify its potential as a therapeutic target in multiple myeloma. Methods： FAS expression was determined by immunohistochemistry, reverse-transcription polymerase chain reaction （RT-PCR） and immunoblot analysis in bone marrow samples obtained from 27 patients with multiple myeloma （MM patients） and peripheral blood mononuclear cells （PBMCs） obtained from 12 healthy donors In parallel, additional analyses were performed on 2 human multiple myeloma cell lines, U266 and RPM18226. U266 cells were treated with cerulenin at various concentrations （5 to 320 μg/ml） for 24 h, and metabolic activity was measured by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assays. Apoptosis was evaluated by dual Annexin V/Pl （propidium iodide） labeling and flow cytometry （FCM） in U266 cells treated with 20 μg/ml cerulenin for 12 h or 24 h. Results： By immunohistochemistry, we found that 19 of 27 bone marrow samples obtained from MM patients expressed significantly high levels of FAS. Similarly, by RT-PCR, 22 of 27 bone marrow samples obtained from MM patients, U266 and RPM18226 showed FAS expression, whereas PBMC samples from 12 healthy donors did not express detectable level of FAS. FAS protein expression was confirmed by immunoblot analysis in 16 of 27 bone marrow samples obtained from MM patients, U266 and RPM18226 cell lines, and no FAS protein expression was detected in PBMC samples from 12 healthy donors. U266 cells were highly sensitive to cerulenin treatment, with a dosage-related effect on metabolic activity, as a measure for cell proliferation. U266 cells treated with 20 μg/ml cerulenin for 12 and 24 h also showed early sign of apoptosis with 56.9% and 69.3% Annexin V^＋/Pl cells, and late apoptotic and necrotic cells with 3.2% and 17.6% Annexin V^＋/Pl^＋ cells. Conclusion： Increased FAS expression existed in multiple myeloma samples and human myeloma cell lines. Cerulenin greatly inhibited metabolic activity/cell proliferation of U266 cells and induced apoptosis, suggesting that FAS is an effective target for pharmacological therapy in human multiple myeloma.
文摘目的研究浅蓝菌素(cerulenin)对人成骨肉瘤细胞U2-OS在裸鼠体内生长抑制作用。方法建立人成骨肉瘤细胞U2-OS BALB/C裸鼠瘤模型。18只成瘤裸鼠按随机数字表分成3组,每组6只;对照组(DMSO溶剂)、低剂量cerulenin组(0.2 mL cerulenin配制液40 mg.kg-1.次-1)、高剂量cerulenin组(0.2 mL cerulenin配制液80 mg.kg-1.次-1)于腹腔隔日注射连续6次。动态观察3组肿瘤体积及裸鼠质量变化,用药结束后18 d处死裸鼠,测量瘤重、计算瘤重抑制率,测量肿瘤体积、计算肿瘤体积抑制率。肿瘤组织行形态学观察。TUNEL法检测肿瘤组织细胞凋亡情况。结果高、低剂量cerulenin组肿瘤体积抑制率分别为51.2%和24.2%,瘤重抑制率分别为49.1%和25.5%。各组间瘤体体积与瘤重比较,差异均有统计学意义(P<0.05)。瘤体组织学显示cerulenin组较对照组出现明显的细胞凋亡的超微结构改变,瘤细胞体积较小,多数细胞微绒毛消失,胞质密度增高。TUNEL法显示高、低剂量cerulenin组,对照组癌细胞凋亡率为(17.1±1.2)%、(10.3±0.8)%、(3.5±0.5)%,3组比较差异均有统计学意义(P<0.05)。结论 cerulenin能够有效诱导人成骨肉瘤细胞U2-OS在裸鼠体内凋亡,抑制肿瘤的生长。
