软骨抗肿瘤制剂的制备及肿瘤实验治疗

小牛软骨经盐酸胍抽提、丙酮分级沉淀及膜超滤等步骤获得了软骨抗肿瘤制剂（Carti-lage Antitumor Preparation, CATP）。利用~3H-TdR参入、测定肿瘤干细胞及细胞死亡率等方法研究了CATP对肿瘤细胞及血管内皮细胞的抑制效应,并对小鼠移植性肿瘤进行了抗肿瘤作用的研究。结果表明:（1）合适浓度的CATP可显著地抑制肿瘤细胞和血管内皮细胞的生长,而不抑制正常细胞;（2）CATP对荷瘤小鼠肿瘤具有明显的抑制作用,抑瘤率为50％～60％。给药组小鼠胸腺重量明显增加,提示CATP可能具有免疫调节作用。实验结果为CATP的临床应用提供了依据。     

鲨鱼软骨抗肿瘤制剂的效应及其作用机制

以鲨鱼软骨为原料，经盐酸胍抽提、丙酮分级沉淀、超滤等步骤得到鲨鱼软骨抗肿瘤制剂（SCATP）。采用放射免疫分析方法测定其对荷瘤小鼠血浆6－Keto－PGF1α和TXB2水平的影响，整装细胞扫描电镜技术测定SCATP对Hela细胞骨架的影响，常规方法测定SCATP对小鼠脾脏和胸腺的影响及对荷瘤小鼠的抑癌效应。结果显示，（1）当剂量达20mg／kg时，SCATP能使荷瘤小鼠血浆6－Keto－PGF1α非常显著下降（P＜0．01），使TXB2水平显著下降（P＜0．05）；（2）SCATP能使Hela细胞骨架发生凝聚或固缩，并有明显的浓度依赖关系，提示SCATP能抑制肿瘤细胞的分裂增殖及运动迁移；（3）SCATP能刺激小鼠脾脏和胸腺，并显著抑制实验动物肿瘤的生长。     

鲨鱼软骨提取物抑瘤作用及其机制研究

目的研究鲨鱼软骨提取物的抗肿瘤作用及其机制.方法采用MTT法及流式细胞仪技术测定鲨鱼软骨粉提取物对血管内皮细胞增殖及产生白细胞介素-6(1L-6)的影响,常规方法测定其对荷S-180A小鼠的抑瘤作用.结果鲨鱼软骨提取物浓度为200～400μg·ml-1时显著抑制内皮细胞的增殖(p<0.05),细胞周期检测发现其主要抑制细胞进入S期;(2)鲨鱼软骨提取物(200～400μg·     

软骨抗癌活性成分抑制血管生成机理的研究

利用从牛软骨中提取的软骨抗肿瘤制剂、软骨血管生成抑制剂和软骨血管生成抑制因子，分别测定了它们对血管内皮细胞DNA合成、鸡胚绒毛尿囊膜血管生成以及内皮细胞迁移运动的抑制效应。提示软骨中提取的这些抗癌活性成分有可能为临床开辟一条以抑制肿瘤血管生成为特色的抗癌治疗新途径。     

鲨骨粉抗癌增效及调节免疫功能的研究

根据祖国医学对鲨鱼药用功效记载,结合现代对鲨鱼软骨的认识,本项研究探讨了鲨鱼软骨抗癌增效及调节免疫功能的作用。结果表明：鲨鱼软骨可明显提高荷瘤小鼠的NK细胞活性和Mψ细胞的吞噬功能。鲨鱼软骨粉与化疗药物（CTX、5Fu、ADM）合用,抗肿瘤（S180、Lewis肺癌）的增效率可达22．8％～47．9％;与放疗（50Co照射）合用,抗肿瘤增效率为1520％～26．50％,证实鲨鱼软骨具有较好的扶正（提高免疫功能）作用和明显的抗癌增效作用。     

鲨鱼软骨制剂的临床应用

观察鲨鱼软骨制剂对多种肿瘤的治疗效果。方法：应用鲨鱼软骨胶囊治疗各种晚期实体瘤８１例,并对乳腺癌和肺癌设治疗对照组,观察其疗效。结果：５２例（６４．２％）患者肿瘤缩小或消失,其中乳腺癌和肺癌的疗效分别为８３．３％和７１．４％,且加用鲨鱼软骨胶囊后常规联合化疗的疗效优于单一化疗的效果。结论：鲨鱼软骨制剂含有高效抗肿瘤血管生长抑制因子,对各种实体肿瘤均有良好的疗效。     

重组人内抑素的抗肿瘤活性

目的观察重组人内抑素抗肿瘤活性及对血管内皮细胞增殖的抑制作用。方法用人癌裸鼠移植性肿瘤及小鼠移植性肿瘤模型对重组人内抑素进行抗肿瘤药效学观察,用MTT法观察其对血管内皮细胞及肿瘤细胞增殖的抑制作用。结果重组人内抑素,可明显抑制人胃癌BGC803和人乳腺癌B37裸鼠移植性肿瘤的生长,对小鼠肝癌H22实体瘤亦呈一定的抑制作用。MTT试验结果显示重组人内抑素可抑制人胎脐静脉血管内皮细胞ECV304的增殖,对人结肠癌HCT-8等肿瘤细胞的增殖无影响。结论重组人内抑素具有较强的抗肿瘤作用,其作用机理可能与抑制血管内皮细胞增殖、抑制肿瘤新生血管形成有关。     

魟鱼软骨多糖对人脐静脉内皮细胞形成新生血管的影响

目的探讨缸鱼软骨多糖(RCG)对人脐静脉内皮细胞形成新生血管的作用.方法原代培养人脐静脉内皮细胞,实验分为生理氯化钠溶液(NS)对照组、RCG 100,50,25,10,2 g·L-1组和阳性对照鲨鱼软骨多糖(SCG,50 g·L-1)组.采用MTT法检测RCG对HUVEC增殖的影响,流式细胞仪检测RCG对HUVEC细胞周期的影响;并观察RCG对HUVEC迁移及小管形成的作用.结果10～100 g·L-1RCG可明显抑制HUVEC的体外增殖,IC50为62.93 g·L-1.流式细胞仪检测表明,RCG阻止HUVEC在G2/M期.10～100g·L-1 RCG明显抑制HUVEC迁移和小管形成.结论RCG具有良好的体外抗血管生成活性.     

鲨骨粉抗吕增效及调节免疫功能的研究

根据祖国医学对鲨鱼药用功效记载,结合现代对鲨鱼软骨的认识,本项研究探讨了鲨鱼软骨抗癌增效及调节免疫功能的作用。结果表明：鲨鱼软骨可明显提高荷瘤小鼠的ＮＫ细胞活性和ＭΦ细胞的吞噬功能。鲨鱼软骨粉与化疗药物（ＣＴＸ、５Ｆｕ、ＡＤＭ）合用,抗肿瘤（Ｓ１８０、Ｌｅｗｉｓ肺癌）的增效率可达２２．８％～４７．９％;与放疗（＾５０Ｃｏ照射）合用,抗肿瘤增效率为１５．２０％～２６．５０％,证实鲨鱼软骨具有较好的扶     

芸香宁碱抑制血管生成作用的研究

目的：研究芸香宁碱对血管生成的抑制作用,并研究其初步的作用机制。方法用人脐静脉内皮细胞（HUVEC）的生长、迁移实验及体内动物模型－鸡胚绒毛尿囊膜法（CAM 法）研究化合物的体内外抗血管生成作用;用酶联免疫吸附法检测芸香宁碱在非凋亡剂量时对肿瘤细胞培养上清液中 VEGF 蛋白分泌量的影响。结果芸香宁碱对血管内皮细胞具有优先抑制作用,对 HUVEC 的半数抑制剂量为（33．2±0．4）μmol·L －1,而对其他肿瘤细胞的 IC50值均大于这一数值;芸香宁碱在体外能够明显抑制内皮细胞的迁移和对细胞外基质黏附作用,并呈剂量依赖性,体内实验显示30μmol·L －1剂量浓度时显著抑制 CAM新生血管形成;芸香宁碱在15μmol·L －1和30μmol·L －1的浓度剂量时显著抑制人肝癌 HepG2细胞培养上清液中VEGF 蛋白的分泌,具有显著性差异。结论芸香宁碱在非凋亡浓度剂量具有抑制新生血管形成作用,其机制与抑制血管内皮细胞增殖以及抑制肿瘤细胞表达 VEGF 有关。     

人肝癌耐药细胞系的建立及生物学特性的研究

目的:培养建立耐药肝癌细胞模型QGY7703/DDP,分析人肝癌耐药细胞系生物学特性的改变.方法:应用人肝癌细胞株QGY7703,采用顺铂(DDP)逐步提高加间歇诱导历时6个月在体外建立QGY7703/DDP细胞系模型.观察该细胞的生长规律,利用四甲基偶氮唑蓝(MTT)法分析暴露前后细胞的增殖和药物对细胞毒性的差异.结果:经过将QGY7703持续暴露,细胞对化疗药物5-氟尿嘧啶(5-FU)、DDP、阿霉素(ADM)、丝裂霉素(MMC)的敏感性发生了改变.QGY7703/DDP对5-FU的耐药性是亲代细胞的1.533倍,对DDP的耐药性是亲代细胞的2.181倍,对ADM的耐药性是亲代细胞的7.080倍,对MMC的耐药性是亲代细胞的9.461倍.结论:化疗药物能够诱导培养的肿瘤细胞产生耐药性.     

赖氨酰谷氨酸二肽的抗肿瘤活性研究

目的研究二肽Lys-Glu(Vilon)的抗肿瘤活性。方法用液相法人工合成了抗肿瘤二肽Vilon,分子量为275.3。用细胞计数法和MTT法测定Vilon对人肠癌LOVO,人胃癌MKN45和人肝癌QGY77033种肿瘤细胞生长的抑制作用,以及对人体正常细胞生长的影响;并对其进行了体内实验。结果Vilon对体外培养的LOVO,MKN45和QGY7703细胞具有剂量依赖性抑制作用,但对人正常白细胞无明显抑制作用。体内抑瘤实验表明,Vilon对小鼠肝癌H22的生长有抑制作用,有效剂量为15mg.kg-1,当使用高剂量30mg.kg-1时,对小鼠移植性肿瘤肝癌H22的抑瘤率达0.60以上,且具有剂效关系。结论Vilon具有明显的体外、体内抗肿瘤活性。     

白细胞介素—1阻滞剂CK—119和CK—122地成纤维样细胞增生的抑制

AIM: To study potent and nontoxic agents to inhibit fibroblast proliferation. METHODS: Fibroblast-like corneal and conjunctival cells were cultured and inhibited by interleukin-1 (IL-1) blockers, dihydropyridazino-pyridazines CK-119 and CK-122. The cell growth and syntheses of DNA, RNA, and protein after IL-1 blocker incubation were determined. RESULTS: CK-119 and CK-122 inhibited cell growth of corneal fibroblast at 30 mg.L-1. DNA and RNA syntheses in corneal fibroblasts were markedly inhibited by CK-119 and CK-122 whereas protein synthesis was either unaffected or mostly enhanced at 30-100 mg.L-1 and 100-300 mg.L-1, respectively. Similar results were obtained in conjunctival cell cultures by CK-119 and CK-122 at 3-10 mg.L-1 and 30-100 mg.L-1, respectively. CONCLUSION: CK-119 and CK-122 are potent IL-1 blockers to inhibit cell growth of fibroblast-like corneal and conjunctival cells mainly through the inhibition of DNA and RNA syntheses but not protein synthesis.     

大蒜素对人宫颈癌Hela细胞增殖影响及其作用机制

目的探讨大蒜素对体外培养宫颈癌Hela细胞增殖的影响及其作用机制。方法采用四甲基偶氮唑蓝（MTT）法测定不同浓度大蒜素对Hela细胞增殖抑制率;流式细胞术测定细胞周期变化及凋亡相关蛋白Bcl-2与Bax的表达。结果 MTT显示大蒜素能抑制人宫颈癌Hela细胞的增殖,并具有时间-剂量依赖性关系;流式细胞术测定结果显示10μg/ml大蒜素可明显诱导宫颈癌Hela细胞凋亡,并将细胞周期阻滞在G2/M期;可降低癌基因蛋白Bcl表达（P〈0.01）,而促进Bax表达（P〈0.01）。结论大蒜素对人宫颈癌Hela细胞增殖具有抑制作用,与细胞周期阻滞和凋亡相关蛋白表达有关。     

Effect of TM208 on QGY-7703 xenograft tumor growth

A newly synthesized dithiocarbamate derivative, 4-methylpiperazine-1-carbodithioc-acid-3-cyano-3,3-diphenylpropyl ester hydrochloride (TM208), has demonstrated anticancer effects with low toxicity in earlier studies; however, the mechanism has yet to be identified. We explored antitumor effects of TM208 and the possible mechanisms by which it inhibited the growth of human hepatocellular carcinoma cell line QGY-7703 xenograft tumors. Cell proliferation was evaluated with the sulforhodamine B assay in vitro. The results suggested that TM208 had slightly antiproliferative activity on QGY-7703 cells. The antitumor effect of TM208 was assessed in nude mice xenografted with QGY-7703 tumors. We found that TM208 significantly inhibited tumor growth but did not cause loss of body weight or leukocytopenia. Western blotting was used to detect the expression of protein kinase C alpha, mitogen-activated protein kinase signal pathways, and cell cycle-related proteins. The results showed that TM208 decreased the expression of protein kinase C alpha, phospho-extracellular signal-regulated kinase-1/2, phospho-p38, cyclin B1, cell division cycle 2 (cdc2), and phospho-cdc2 (Thr161) and increased the expression of phospho-cdc2 (Tyr15). Taken together, our data show that TM208 has little antiproliferative effect on QGY-7703 cells in vitro, whereas it significantly inhibits the growth of QGY-7703 xenograft tumors with low toxicity in vivo. The inhibition of mitogen-activated protein kinase signal pathways and the regulation of the G2/M phase may be responsible for its antitumor effects.     

黄芪总苷的抑瘤作用及其作用机制

目的　探讨黄芪总苷的抗肿瘤作用及其作用机制。方法　采用小鼠肝癌 (HepA)和肉瘤 (S1 80 )两种小鼠移植瘤的动物模型 ,以瘤重抑制率作指标。体外采用人宫颈癌细胞株HeLa细胞 ,用MTT法测肿瘤细胞的生长 ,流式细胞术及TUNEL法检测细胞周期及细胞凋亡。结果　黄芪总苷显著抑制小鼠肝癌 (HepA)和肉瘤 (S1 80 )的生长 ;体外可显著抑制HeLa细胞的生长 ,使细胞周期阻滞于G0 /G1 期 ,并诱导其凋亡。结论　黄芪总苷对小鼠肝癌 (HepA)与肉瘤(S1 80 )具有抑瘤作用。对HeLa细胞的生长有直接抑制作用。其抗肿瘤作用可能与细胞周期阻滞于G0 /G1 期和诱导细胞凋亡有关     

Inhibitory effects of artesunate on angiogenesis and on expressions of vascular endothelial growth factor and VEGF receptor KDR/flk-1

Artesunate (ART) is a semi-synthetic derivative of artemisinin extracted from the plant Artemisia annua is a safe and effective antimalarial drug. In the present investigation, ART was found also to inhibit angiogenesis in vivo and in vitro. The anti-angiogenic effect in vivo was evaluated in nude mice by means of human ovarian cancer HO-8910 implantation and immunohistochemical stainings for microvessel (CD(31)), vascular endothelial growth factor (VEGF) and VEGF receptor KDR/flk-1. Tumor growth was decreased and microvessel density was reduced following drug treatment with no apparent toxicity to the animals. ART also remarkably lowered VEGF expression on tumor cells and KDR/flk-1 expression on endothelial cells as well as tumor cells. The in vitro effect of ART was tested on models of angiogenesis, namely, proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVEC). The results showed that ART significantly inhibited angiogenesis in a dose-dependent form in the range of 0.5 approximately 50 micromol/l. Additionally, the inhibitory effect of ART on HVUEC proliferation was stronger than that on Hela, JAR, HO-8910 cancer cells, NIH-3T3 fibroblast cells and human endometrial cells, indicating that ART was selectively against HUVEC. These findings and the known low toxicity of ART are clues that ART may be a promising angiogenesis inhibitor.     

姜黄素对人宫颈癌Hela细胞增殖的抑制作用

目的:研究姜黄素对人宫颈癌Hela细胞生长的抑制作用。方法:姜黄经75%乙醇提取、纯化得姜黄素。MTT法检测姜黄素对体外培养Hela细胞增殖的抑制作用;倒置显微镜观察Hela细胞形态学变化;Western blot法检测抑癌基因p53蛋白的表达;流式细胞仪检测Hela细胞凋亡和周期分布。结果:姜黄素对Hela细胞生长24 h的最佳抑制质量浓度为116.68μg/ml,抑制率为63.20%;11.67、35.00、58.34μg/ml姜黄素培养24 h,人宫颈癌Hela细胞数目明显减少,细胞变圆、缩小、老化,核质发散,并与质量浓度呈正相关。29.17、145.85μg/ml姜黄素可增强p53蛋白的表达;11.67、35.00、58.34μg/ml姜黄素可促进人宫颈癌Hela细胞凋亡,并与质量浓度呈正相关,G0/G1期细胞和S期细胞逐渐减少,G2/M期细胞逐渐增多。结论:姜黄素对人宫颈癌Hela细胞增殖有明显抑制作用,并能促进其凋亡和p53的表达,阻滞Hela细胞的G2/M期。     

Zerumbone,Sesquiterpene Photochemical from Ginger,Inhibits Angiogenesis

Here, we investigated the role of zerumbone, a natural cyclic sesquiterpene of Zingiber zerumbet Smith, on angiogenesis using human umbilical vein endothelial cells (HUVECs). Zerumbone inhibited HUVECs proliferation, migration and tubule formation, as well as angiogenic activity by rat aorta explants. In particular, zerumbone inhibited phosphorylation of vascular endothelial growth factor receptor-2 and fibroblast growth factor receptor-1, which are key regulators of endothelial cell function and angiogenesis. In vivo matrigel plug assay in mice demonstrated significant decrease in vascularization and hemoglobin content in the plugs from zerumbone-treated mice, compared with control mice. Overall, these results suggest that zerumbone inhibits various attributes of angiogenesis, which might contribute to its reported antitumor effects.     

Valproic acid inhibits angiogenesis in vitro and in vivo

Valproic acid (VPA) is a widely used antiepileptic agent that is undergoing clinical evaluation for anticancer therapy. We assessed the effects of VPA on angiogenesis in vitro and in vivo. In human umbilical vein endothelial cells, therapeutically relevant concentrations of VPA (0.25 to 1 mM) inhibited proliferation, migration, and tube formation. VPA 1 mM inhibited endothelial cell proliferation by 51 +/- 5%, migration by 86 +/- 11%, and tube formation by 82 +/- 3%. These changes were preceded by the hyperacetylation of histone H4, indicating the inhibition of histone deacetylase (HDAC), and a decreased expression of the endothelial nitric-oxide synthase (eNOS). The inhibition of endothelial cell tube formation by VPA was prevented by addition of the nitric oxide donor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA NONOate). The anticonvulsive active VPA derivative 2-ethyl-4-methylpentanoic acid, which does not inhibit HDAC, did not affect endothelial cell proliferation, tube formation, or eNOS expression. VPA was also found to inhibit angiogenesis in vivo in the chicken chorioallantoic membrane assay and in a Matrigel plug assay in mice. Embryos from VPA-treated mice showed disturbed vessel formation. These results indicate that therapeutic plasma levels of VPA inhibit angiogenesis by a mechanism involving a decrease in eNOS expression preceded by HDAC inhibition.