Yeh J Y, Huang W J, Kan S F, Wang P S
Department and Graduate Institute of Physiology, School of Life Science, National Yang-Ming University, Taipei, Taiwan, Republic of China.
J Urol. 2001 Nov;166(5):1937-42.
Digitalis or cardiac glycosides have been noted to induce tumor static or oncolytic effects in various types of cancer. We evaluated the effects and underlying mechanisms of cardiac glycosides, including digoxin, digitoxin and ouabain, on the proliferation of hormone dependent and independent prostate cancer cell lines.
Cell proliferation of the 3 human prostate cancer cell lines LNCaP, DU145 and PC3 was measured by 3-(4,5-dimethylthiazol-2-yle)2,5-diphenyltetralozium bromide (Sigma Chemical Co., St. Louis, Missouri) colorimetric assay. The cytotoxic effects of digitalis on prostate cancer cells were determined by lactate dehydrogenase measurements of the culture medium. Intracellular Ca2+ was measured by a dual wavelength spectrometer system. The percent of apoptotic cells after digitalis treatment was measured by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling and flow cytometry.
Digoxin, digitoxin and ouabain significantly inhibited the proliferation of LNCaP, DU145 and PC3 cells at a dose of 1 or 10 microM. after 1 to 4 days of culture. Cytotoxicity of digitalis on the DU145 and LNCaP cells was dose dependent but cytotoxicity was not obvious in PC3. Digitalis (1 microM.) significantly increased intracellular Ca2+ in LNCaP and DU145 after 12 hours of culture but PC3 cells needed a 24-hour treatment to show any effect. In the apoptosis measurement digitalis at a dose of 1 and 10 microM. also significantly increased the percent of apoptotic cells in the LNCaP, DU145 and PC3 cell lines. Normal control human glomerular epithelial cells showed no response to digitalis treatment at all tested doses.
Digitalis may inhibit the proliferation of prostate cancer cell lines, although the 3 cell lines showed varied sensitivity to digitalis. These effects are possibly the result of a mechanism involving sustained elevation of the concentration of intracellular Ca2+ and of apoptosis.
已注意到洋地黄或强心苷可在多种类型癌症中诱导肿瘤静止或溶瘤效应。我们评估了包括地高辛、洋地黄毒苷和哇巴因在内的强心苷对激素依赖性和非依赖性前列腺癌细胞系增殖的影响及潜在机制。
通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（西格玛化学公司，密苏里州圣路易斯）比色法测定3种人前列腺癌细胞系LNCaP、DU145和PC3的细胞增殖。通过测定培养基中的乳酸脱氢酶来确定洋地黄对前列腺癌细胞的细胞毒性作用。用双波长光谱仪系统测量细胞内钙离子浓度。通过末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记和流式细胞术测量洋地黄处理后凋亡细胞的百分比。
地高辛、洋地黄毒苷和哇巴因在1或10微摩尔剂量下，培养1至4天后可显著抑制LNCaP、DU145和PC3细胞的增殖。洋地黄对DU145和LNCaP细胞的细胞毒性呈剂量依赖性，但对PC3细胞的细胞毒性不明显。培养12小时后，1微摩尔的洋地黄可显著增加LNCaP和DU145细胞内的钙离子浓度，但PC3细胞需要24小时处理才会显示出任何效应。在凋亡检测中，1和10微摩尔剂量的洋地黄也显著增加了LNCaP、DU145和PC3细胞系中凋亡细胞的百分比。在所有测试剂量下，正常对照人肾小球上皮细胞对洋地黄处理均无反应。
洋地黄可能抑制前列腺癌细胞系的增殖，尽管这3种细胞系对洋地黄表现出不同的敏感性。这些效应可能是由涉及细胞内钙离子浓度持续升高和凋亡的机制导致的。
