三尖杉酯碱和高三尖杉酯碱对肿瘤细胞Ⅳ型胶原酶的分泌及活性的影响

目的：植物来源的抗白血病药三尖杉酯碱和高三尖杉酯碱对肿瘤细胞增殖有显抑制，已成为急性粒细胞性自血病的首选药物之一：我们的研究还表明该药对肿瘤细胞的侵袭和运动也有明显抑制作用。为探讨其作用机理，进一步观察了该药对人纤维肉瘤HT-1080细胞Ⅳ型胶原酶的分泌及活性的影响。方法：用聚丙烯酰胺凝胶电泳法测定HT-1080细胞Ⅳ型胶原酶MMP-2和MMP-9的产生及活性。结果：三尖杉酯碱在0．001，0．01μg/ml浓度下对HT-1080细胞MMP-2和MMP-9的产生及活性均无明显影响。     

三尖杉酯碱和高三尖杉酯碱诱导人早幼粒白血病细胞的程序性死亡

体外实验证明2×1O^-7 mol·L^-1 三尖杉酯碱和 1O^-7 mol·L^-1高三尖杉酯碱与HL-60细胞作用 4 h，可诱导该细胞产生程序性死亡。提取细胞DNA进行琼脂糖凝胶电泳呈现典型的DNA 梯，同时出现细胞核染色体断裂、核固缩及细胞空泡样变等形态变化。并且在一定范围内具有作用时间和浓度依赖性，二者诱导HL-60细胞程序性死亡作用的强弱与其细胞毒作用平行。     

小鼠造血干细胞和P388白血病干细胞对5种抗癌药的敏感性比较

用脾集落形成法比较了5种抗癌药对小鼠造血干细胞(NCFU-S)和P₃₈₈白血病干细胞(LCFU-S)的作用。三尖杉酯碱、半合成三尖杉酯碱、高三尖杉酯碱和环磷酰胺对两类干细胞作用的剂量—反应曲线呈指数形,效能比依次为5.15,6.01,7.96和9.98。阿糖胞苷对NCFU-S无明显作用,但对LCFU-S杀伤作用强,大剂量时剂量—反应曲线趋于水平。当三尖杉酯碱、高三尖杉酯碱和半合成三尖杉酯碱的剂量分别低于0.30,0.28和0.75mg/kg时,对LCFU-S无明显作用,剂量—反应曲线上出现“肩形”,提示LCFU-S在小剂量三尖杉酯类生物碱作用下,有一受亚致死性损伤后修复过程。     

三尖杉酯碱对肿瘤细胞核苷酸代谢的影响

本文用标记核苷参入艾氏腹水癌细胞各成分以研究三尖杉酯碱对有关的核苷酸代谢的影响。结果表明,该药在抑制DNA合成的同时,使³H-胸腺嘧啶核苷参入酸溶部分明显增加,其中以³H-胸苷三磷酸的增加最为显著。³H-胸苷-磷酸、³H-胸苷二磷酸也有明显增加;但各胸腺嘧啶核苷酸占酸溶部分掺入的百分比与对照组相同。说明三尖杉酯碱不抑制胸腺嘧啶核苷的磷酸化,而抑制脱氧核苷三磷酸到DNA的聚合过程。在³H-尿嘧啶核苷和³H-次黄嘌呤参入实验中,三尖杉酯碱对四种³H-核苷三磷酸及RNA的合成无明显影响。综合核苷三磷酸及脱氧核苷三磷酸的测定结果,表明该药似乎对核糖核苷二磷酸的还原无抑制作用。     

小鼠造血干细胞和P_(388)白血病干细胞对5种抗癌药的敏感性比较

用脾集落形成法比较了5种抗癌药对小鼠造血干细胞(NCFU-S)和P_(388)白血病干细胞(LCFU-S)的作用。三尖杉酯碱、半合成三尖杉酯碱、高三尖杉酯碱和环磷酰胺对两类干细胞作用的剂量—反应曲线呈指数形,效能比依次为5.15,6.01,7.96和9.98。阿糖胞苷对NCFU-S无明显作用,但对LCFU-S杀伤作用强,大剂量时剂量—反应曲线趋于水平。当三尖杉酯碱、高三尖杉酯碱和半合成三尖杉酯碱的剂量分别低于0.30,0.28和0.75mg/kg时,对LCFU-S无明显作用,剂量—反应曲线上出现“肩形”,提示LCFU-S在小剂量三尖杉酯类生物碱作用下,有一受亚致死性损伤后修复过程。     

高三尖杉酯碱抗乳腺癌作用的初步研究

目的:研究药物高三尖杉酯碱是否对人乳腺癌MDA-MB-453细胞具有生长抑制和诱导凋亡作用,以挖掘研发治疗乳腺癌的新药物.方法:常规培养人乳腺癌MDA-MB-453细胞,根据高三尖杉酯碱浓度分为分为组和高三尖杉酯碱10、20、40、60、80、120、160μg/mL组,其中20μg/mL组为小剂量组,80μg/mL组为中剂量组,120 μg/mL组为大剂量组.采用台盼蓝染色、MTT比色法和透射电镜分析研究高三尖杉酯碱对于人乳腺癌细胞的作用.结果:台盼蓝染色结果显示大、中、小剂量组的活细胞率依次由小到大;MTT比色法显示各浓度药物作用在24、48和72 h均能显著抑制人乳腺癌MDA-MB-453细胞的增殖(P<0.05,P<0.01).电镜观察发现,高三尖杉酯碱作用后的人乳腺癌MDA-MB-453细胞染色体断裂,核分裂成碎片,细胞内形成凋亡小体.结论:高三尖杉酯碱对于乳腺癌MDA-MB-453细胞具有一定的抑制作用和诱导凋亡的作用,应予进一步研究.     

高三尖杉酯碱抗乳腺癌作用的初步研究

目的:研究药物高三尖杉酯碱是否对人乳腺癌MDA-MB-453细胞具有生长抑制和诱导凋亡作用,以挖掘研发治疗乳腺癌的新药物.方法:常规培养人乳腺癌MDA-MB-453细胞,根据高三尖杉酯碱浓度分为分为组和高三尖杉酯碱10、20、40、60、80、120、160μg/mL组,其中20μg/mL组为小剂量组,80μg/mL组为中剂量组,120 μg/mL组为大剂量组.采用台盼蓝染色、MTT比色法和透射电镜分析研究高三尖杉酯碱对于人乳腺癌细胞的作用.结果:台盼蓝染色结果显示大、中、小剂量组的活细胞率依次由小到大;MTT比色法显示各浓度药物作用在24、48和72 h均能显著抑制人乳腺癌MDA-MB-453细胞的增殖(P<0.05,P<0.01).电镜观察发现,高三尖杉酯碱作用后的人乳腺癌MDA-MB-453细胞染色体断裂,核分裂成碎片,细胞内形成凋亡小体.结论:高三尖杉酯碱对于乳腺癌MDA-MB-453细胞具有一定的抑制作用和诱导凋亡的作用,应予进一步研究.     

抗肿瘤植物药的非抗肿瘤临床应用

<正> 抗肿瘤植物药是抗肿瘤药物的重要组成部分。已有的植物类抗肿瘤药在肿瘤的治疗中发挥着一定作用,而且在其他疾病尤其是疑难杂病治疗中显示出活力。本文就抗肿瘤药物三尖杉酯碱类、长春碱类、秋水仙碱类、鬼臼毒素、喜树碱和紫杉醇在临床中用于治疗非肿瘤疾病的现状作一综述。1 三尖杉酯碱和高三尖杉酯碱 三尖杉酯碱的早期研究证明,它可抑制蛋白质合成的起始阶段,以后的进一步研究表明,它对DNA合成也有抑制作用,还能诱导细胞分化,提高胞内cAMP含量。高三尖杉酯碱作用基本上与三尖杉酯碱相同,临床证明两药均对急性非淋巴性白血病有较高疗效。两药的非抗肿瘤作用主要有:(1)防治青光眼     

人白血病HL60细胞的分化状态对细胞凋亡的影响

用细胞培养和流式细胞术等方法,研究人白血病HL60细胞诱导分化后,对三尖杉酯碱(Har)和喜树碱(Cam)诱导细胞凋亡的影响。结果表明,12-豆蔻酰及13-乙酸佛波酯以16nmol·L^-1浓度处理HL60细胞24h,细胞向单核/巨噬细胞方向分化,阻断于G_{1期;分化细胞抗Har和Cam诱导的细胞凋亡,但其c-myc基因的表达无变化。1.4%二甲基亚砜处理HL60细胞48h,细胞向粒细胞方向分化,阻断于G1期;分化细胞抗Cam,而不抗Har诱导的细胞凋亡;分化细胞的c-myc基因表达明显下降。结果提示,人白血病HL60细胞的分化状态,明显影响三尖杉酯碱和喜树碱诱导的细胞凋亡,但可能与c-myc基因的表达变化无关。}     

半合成三尖杉酯碱的抗肿瘤作用及毒性研究

三尖杉酯碱Harringtonine是从我国特有植物海南粗榧(Cephalotaxus hainanensisLi)中分离出的一种抗肿瘤有效成分。对多种动物移植性肿瘤有明显的抗肿瘤作用。临床已经证明该酯碱对多种急性非淋巴性白血病及慢性粒细胞白血病有较好疗效。由于三尖杉     

Magnolol has the ability to induce apoptosis in tumor cells

We previously found that multiple intraperitoneal administration of magnolol from Magnolia obovata inhibited tumor metastasis and growth in vivo, and that the anti-metastatic effect of magnolol was due to the inhibition of the tumor cell invasion. The purpose of this study was to clarify the inhibitory mechanism of magnolol on the growth of tumor cells, and we expect that magnolol may have the ability to induce apoptosis in tumor cells. In an in vitro proliferation assay, 100 microM of magnolol inhibited the proliferation of B16-BL6, THP-1, BAE and HT-1080 cells, but 30 microM of magnolol did not affected cells proliferation. In addition, 100 microM of magnolol induced apoptotic cell death within 24 h in three tumor cell lines, B16-BL6, THP-1 and HT-1080, not BAE cells, and then up-regulated the activity of caspase-3 and caspase-8. The up-regulation of caspases activity by 100 microM of magnolol was suppressed by the inhibitor of all caspases, z-VAD-fmk. These data suggest that magnolol possesses ability to inhibit tumor growth, and the ability is due to the induction of apoptosis with the activation of caspases.     

Anti-invasion and anti-angiogenesis effect of taxol and camptothecin on melanoma cells

Two highly invasive melanoma cell lines B16BL6 and B16F10 were used to investigate the anti-invasion and anti-angiogenesis action of taxol and camptothecin (CPT). The adhesion of melanoma cells was tested by optical absorbance at 545 nm. The invasive activity of these cells was tested in a transwell cell chamber assay. The cell migration within a 3D collagen matrix was recorded with a time-lapse video recorder and analyzed by computer-assisted cell tracking. Gelatin zymography was used to study the metalloproteinase activity. The chicken chorioallantoic membrane (CAM) model was used to study the anti-angiogenesis effect of the two drugs. The results demonstrated that both taxol and CPT could inhibit the migration of B16F10 cells, and inhibit the adhesion of B16F10 to fibronectin and laminin. They can reduce the metalloproteinase secretion of HT1080 and exhibit the anti-angiogenesis effect in the CAM model. Taxol showed a highly anti-invasion effect on B16BL6 cells while CPT did not exhibit such an effect.     

Antiproliferative activity of cardenolides isolated from Streptocaulon juventas

Sixteen cardenolides, two hemiterpenoids, two phenylpropanoids and a phenylethanoid isolated from the roots of Streptocaulon juventas (LOUR.) MERR. were examined for their antiproliferative activity toward three human-derived (HT-1080 fibrosarcoma, lung A549 adenocarcinoma, cervix HeLa adenocarcinoma) and three murine-derived (colon 26-L5 carcinoma, Lewis lung carcinoma, B16-BL6 melanoma) cell lines. The cardenolides selectively and strongly inhibited proliferation of the HT-1080 (IC(50) values, 0.054-1.6 microM) and A549 (IC(50), 0.016-0.65 microM) cell lines. The characteristic morphological changes and ladder-like DNA fragmentation in those cells treated with the cardenolides indicated the antiproliferative activity was due to the induction of apoptosis.     

Antiproliferative activity of Vietnamese medicinal plants

Methanol, methanol-water (1:1) and water extracts were prepared from seventy-seven Vietnamese medicinal plants and tested for their antiproliferative activities against human HT-1080 fibrosarcoma cells. Among them, fifteen extracts including seven methanol extracts of Caesalpinia sappan, Catharanthus roseus, Coscinium fenestratum, Eurycoma longifolia, Hydnophytum formicarum and Streptocaulon juventas (collected at two areas), six methanol-water (1:1) extracts of Cae. sappan, Cat. roseus, Co. fenestratum, H. formicarum and S. juventas (at two areas), and two water extracts of Cae. sappan and S. juventas exhibited antiproliferative activities in a concentration-dependent manner. Their antiproliferative activities against human cervix HeLa adenocarcinoma, human lung A549 adenocarcinoma, murine colon 26-L5 carcinoma, murine Lewis lung carcinoma (LLC) and murine B16-BL6 melanoma cells were then examined. Co. fenestratum showed selective activity against lung carcinoma and/or lung metastatic cell lines, A549, LLC and B16-BL6, while H. formicarum and S. juventas showed selective activity against human tumor cell lines, HeLa and A549. Characteristic morphological change and DNA fragmentation indicated the antiproliferative activity to be due to the induction of apoptosis.     

三尖杉酯类生物硷对小鼠白血病P388及L615蛋白质生物合成的影响

本文初步观察了四种三尖杉酯硷对³H-DL-亮氨酸、³H-L-门冬酰胺、¹⁴C-DL-苯丙氨酸掺入小鼠白血病P₃₈₈腹水细胞及L₆₁₅。脾组织蛋白质的作用。实验结果表明,在等毒性剂量(1/5 LD₅₀)下,一次腹腔(或静脉)给药,四种酯硷对P₃₈₈腹水细胞及L₆₁₅脾组织蛋白质生物合成抑制明显,作用较快,并具有相似的时间—反应曲线。一般在24小时基本恢复。     

Antitumor activity of the Korean Mistletoe Lectin is attributed to activation of macrophages and NK cells

Inhibitory effect of the lectins (KML-C) isolated from Korean mistletoe (KM; Viscum album coloratum) on tumor metastases produced by murine tumor cells (B16-BL6 melanoma, colon 26-M3.1 carcinoma and L5178Y-ML25 lymphoma cells) was investigated in syngeneic mice. An intravenous (i.v.) administration of KML-C (20-50 ng/mouse) 2 days before tumor inoculation significantly inhibited lung metastases of both B16-BL6 and colon 26-M3.1 cells. The prophylactic effect of 50 ng/mouse of KML-C on lung metastasis was almost the same with that of 100 microg/mouse of KM. Treatment with KML-C 1 day after tumor inoculation induced a significant inhibition of not only the experimental lung metastasis induced by B16-BL6 and colon 26-M3.1 cells but also the liver and spleen metastasis of L5178Y-ML25 cells. Furthermore, multiple administration of KML-C given at 3 day-intervals after tumor inoculation led to a significant reduction of lung metastasis and suppression of the growth of B16-BL6 melanoma cells in a spontaneous metastasis model. In an assay for natural killer (NK) cell activity, i.v. administration of KML-C (50 ng/mouse) significantly augmented NK cytotoxicity against Yac-1 tumor cells 2 days after KML-C treatment. In addition, treatment with KML-C (50 ng/mouse) induced tumoricidal activity of peritoneal macrophages against B16-BL6 and 3LL cells. These results suggest that KML-C has an immunomodulating activity to enhance the host defense system against tumors, and that its prophylactic and therapeutic effect on tumor metastasis is associated with the activation of NK cells and macrophages.     

Cordycepin (3′-deoxyadenosine) inhibits the growth of B16-BL6 mouse melanoma cells through the stimulation of adenosine A3 receptor followed by glycogen synthase kinase-3β activation and cyclin D1 suppression

Cordyceps sinensis, a parasitic fungus on the larvae of Lepidoptera, has been used as a traditional Chinese medicine. We previously reported that the growth of B16-BL6 mouse melanoma (B16-BL6) cells was inhibited by cordycepin (3'-deoxyadenosine), an active ingredient of C. sinensis, and its effect was antagonized by MRS1191, a selective adenosine A3 receptor antagonist. In this study, the radioligand binding assay using [125I]-AB-MECA (a selective adenosine A3 receptor agonist) has shown that B16-BL6 cells express adenosine A3 receptors and that cordycepin binds to these receptors. We also confirmed the involvement of adenosine A3 receptors in the action of cordycepin using MRS1523 and MRS1220, specific adenosine A3 receptor antagonists. Next, indirubin, a glycogen synthase kinase-3beta (GSK-3beta) inhibitor, antagonized the growth suppression induced by cordycepin. Furthermore, the level of cyclin D1 protein in B16-BL6 cells was decreased by cordycepin using Western blot analysis. In conclusion, this study demonstrated that cordycepin inhibits the proliferation of B16-BL6 cells by stimulating adenosine A3 receptors followed by the Wnt signaling pathway, including GSK-3beta activation and cyclin D1 inhibition.     

Inhibitory effect of magnolol and honokiol from Magnolia obovata on human fibrosarcoma HT-1080. Invasiveness in vitro.

We investigated the inhibitory effect of Magnolia obovata Thunb. bark ethanol extracts on human fibrosarcoma HT-1080 cells invasion in a reconstituted basement membrane [Matrigel (MG)]. We found that the effective components of the bark ethanol extracts were magnolol and honokiol, two biphenyl compounds. The extracts, magnolol and honokiol, did not affect HT-1080 cells adhesion to MG, but did inhibit HT-1080 cells migration at a high concentration (100 microM). HT-1080 cells secrete matrix metalloproteinase (MMP)-9, which degrades the extracellular matrix as a part of the invasive process. Magnolol and honokiol inhibited the activity of MMP-9, which may have been responsible, in part, for the inhibition of tumor cell invasiveness.     

三尖杉酯类生物碱对L615及P388白血病细胞cAMP含量的影响

实验证明,三尖杉酯类生物碱对某些动物移植性白血病和实体瘤有不同程度的疗效。在治疗剂量下,三尖杉酯碱能抑制肿瘤细胞有丝分裂,降低肝、脾和肿瘤组织的核酸含量,明显抑制DNA合成。三尖杉酯碱对白血病L1210杀伤动力学研究表明,它是一个细胞周期非特异性抗肿瘤药,但对S期细胞有强烈杀伤作用,对G_0期细胞也有一定影响。临床