C_(60)对小鼠S_(180)肉瘤光动力学作用模型的建立

为验证C60 对活体肿瘤的光动力学损伤作用,我们从两方面进行实验 :C60 对荷瘤小鼠的S180实体瘤的光动力学杀伤作用和C60 对离体S180 肿瘤细胞的光动力学杀伤作用。在小鼠的瘤体上注射C60 光敏剂 ,在511nm和578nm混合黄绿色激光照射下 ,激发C60,产生大量的单线态氧 ,杀伤活性肿瘤。在激光光强为500mW ,C60 浓度为30μg/ml时 ,荷瘤小鼠寿命平均延长5天 ,瘤径减小1cm,瘤重减轻0.8克 ,在活体水平上验证了C60 的光动力学作用 ;另外,在离体的S180 肿瘤细胞中加入细胞培养物和C60 脂质体光敏剂 ,用碘钨灯照射激发C60。经MTT法和FDA -PI双色荧光分光光度法检测 ,离体S180 肿瘤细胞也明显被杀伤。C60 具有强的光敏剂作用 ,它不仅在活体水平上可以杀伤S180 实体瘤而且对离体的S180 实体瘤细胞有强的杀伤作用。     

双歧杆菌及其WPG对S180荷瘤小鼠免疫调节和抑瘤的作用研究

目的 通过观察双歧杆菌及其细胞壁肽多糖（Cell Wall Preparation,whole peptidoglycon,WPG)对S180荷瘤小鼠抑瘤作用及在体内外对IL－6和TNF－α生态的影响,探讨双歧杆菌及其WOG的免疫调节和抑瘤作用机制。方法 采用经驯化而具有一定耐氧能力的两歧双歧杆菌C149株及其WPG腹腔免疫S180荷瘤小鼠,应用放射免疫检测小鼠外周血中的IL－6和TNF－α的含量,同时在体外观察小鼠腹腔巨噬细胞产生IL－6和TNF－α的影响。结果 双歧杆菌及其WPG在体内外对IL－6和TNF－α的生成有明显的促进作用,对S180荷瘤小鼠均有明显抑瘤作用。结论 双歧杆菌及其WPG可能通过刺激小鼠的巨噬细胞产生一些免疫活性因子而间接发挥抑瘤作用。     

灵芝中有效成分灵芝酸的抑制肿瘤作用研究

本文采用了体外肿瘤细胞培养增殖抑制试验与体内肿瘤细胞抑制试验,结果显示灵芝酸在体外对肿瘤细胞株SW620、LS180、S180的增殖具有明显的抑制作用,体内抗肿瘤疗效试验显示灵芝酸对Lewis肺癌(足趾接种)具有一定的疗效,对荷Lewis肺癌的小鼠IL-2的生成及NK细胞的免疫活性均有一定的促进作用。因此可以认为灵芝酸通过直接细胞毒作用与激活免疫体系实现抑制肿瘤作用。     

仙茅多糖对氟尿嘧啶增效减毒作用

本文探讨了仙茅多糖(COP)对氟尿嘧啶的增效减毒作用。实验中建立小鼠S180实体瘤模型,连续给药10 d,测定小鼠瘤重,体质量、白细胞、红细胞、胸腺指数、脾脏指数以及小鼠脾脏中的超氧化物歧化酶活性和丙二醛含量。同时建立小鼠S180腹水瘤模型,连续给药30 d,记录小鼠存活天数,计算生命延长率。结果显示,仙茅多糖能改善氟尿嘧啶引起的小鼠白细胞减少、胸腺萎缩、脾脏肿大(P0.05或P0.01);但联用高剂量组使小鼠脾脏中SOD活力(P0.01)降低,联用低、中剂量组使MDA含量增加(P0.05)。COP和5-FU均延长荷瘤小鼠的生存期,(5-FU+COP)联合用药组生命延长率比5-FU单独用药组明显增高,且与联用的COP剂量呈量效关系。从而表明仙茅多糖对氟尿嘧啶具有一定的增效减毒作用,机制可能与其能调节机体免疫力有关,而与过氧化反应无关。     

骨骼肌条件培养液对恶性肿瘤抑制作用的初步研究

目的：研究骨骼肌微环境因素对恶性肿瘤细胞增殖的影响,评价其在骨骼肌转移瘤罕见性中的意义。方法：原代培养新生Wistar大鼠骨骼肌细胞,用MTT法分析、比较其条件性上清液（MMCM）对恶性肿瘤细胞（小鼠实体瘤-肉瘤、细胞株S180-V、小鼠骨髓瘤细胞株SP2／0、人食管癌细胞Eca109）及正常非肿瘤细胞（金黄地鼠肾细胞株BHK-21）的体外抑瘤作用。结果：MMCM与S180—V、SP2／0、Eca109及BHK-21共同培养后,三株恶性肿瘤细胞株的增殖均显著受抑,而正常非肿瘤细胞株则不受影响。结论：新生大鼠骨骼肌细胞可产生选择性抑制恶性肿瘤细胞增殖的抑瘤物质,这种抑瘤物质可能是临床上骨骼肌内罕见转移瘤现象的生物学基础。     

香菇C91-3菌丝发酵蛋白对H22肿瘤细胞体内外抗肿瘤机制的初探

目的探讨香菇C91-3菌丝发酵蛋白(LFP91-3)对H22肿瘤细胞抑瘤作用及相关机制。方法应用不同剂量LFP91-3(50、100和150μg)对H22荷瘤小鼠腹水瘤模型和腋下实体瘤进行治疗,观察其生存期和体内抑瘤作用;并用MTT法(LFP91-3浓度5、10、15μg/mL)和流式细胞仪对经LFP作用的H22肿瘤细胞进行观察和检测。结果香菇C91-3菌丝发酵提取蛋白(LFP91-3)能延长H22荷瘤小鼠的生存期,对体外培养的H22有直接杀伤作用,抑瘤率出现对浓度和时间的依赖性。LFP91-3能将H22肿瘤细胞株细胞周期阻滞到S期并诱导出细胞凋亡。结论 LFP91-3在体内、外对H22肿瘤细胞有较好的抑瘤作用,主要是诱导其调亡。     

桦木酮酸对肿瘤细胞SGC-7901、HepG-2及S180荷瘤小鼠肿瘤细胞的影响

为研究桦木酮酸体外对SGC-7901、HepG-2及体内对S180荷瘤小鼠的影响,采用MTT与肿瘤细胞集落形成能力实验观察桦木酮酸对HepG-2和SGC-7901作用。通过抑瘤率观察其对S180荷瘤小鼠的影响;HE染色观察其对S180肿瘤细胞形态学的影响。结果显示：桦木酮酸可抑HepG-2和SGC-7901细胞的生长,MTT与肿瘤集落形成能力实验测得其IC50分别为68．14和110．77μmol／L。连续给药8d后,150和75ms／kg／d剂量的桦木酮酸对S180的抑制率分别为68．3％和41．5％（P〈0．05）;HE染色发现肿瘤细胞表现较为明显的胀亡现象：细胞淡染、细胞体积增大,细胞核结构相对完整。提示桦木酮酸体内、外对肿瘤细胞均有一定的抑制作用,其作用可能与其引起肿瘤细胞胀亡有关。     

PVAX-MAGE-1基因疫苗延缓小鼠肝癌细胞H22腹水瘤和实体瘤的生长

为观察重组基因疫苗PVAX-MAGE-1的抑瘤效应,构建黑色素瘤抗原-1(melanoma antigen-1,MAGE-1)真核基因表达载体--PVAX-MAGE-1.以重组质粒免疫C57BL/6小鼠后,ELISA法检测表明,与对照鼠(PVAX-1和生理盐水注射小鼠)比较,免疫小鼠脾淋巴细胞上清液中的细胞因子IL-2和IFN-γ明显升高(P0.05);淋巴细胞-肿瘤细胞混合培养证明,免疫小鼠外周血CD8+T细胞对靶细胞的特异性杀伤作用明显增强(P0.05).体内实验证明,PVAX-MAGE-1免疫C57BL/6小鼠,可显著延缓移植性H22腹水瘤及实体瘤在小鼠体内的生长.实验结果提示,重组基因疫苗PVAX-MAGE-1有明显的延缓肿瘤生长的作用,其抑瘤作用与提高T淋巴细胞IL和IFN表达,增强对肿瘤杀伤作用直接相关.     

灵芝发酵液抗肿瘤的初步研究

对灵芝发酵液抑瘤作用做初步研究。研究表明灵芝发酵液（GLF）能显著延长腹水瘤小鼠的生存期（P＜0．005）,对实体瘤具有显著抑制作用,抑瘤率为64．84％（P＜0．01）。灵芝发酵液体外对3种瘤细胞无直接杀伤作用（P＜0．05）。分析灵芝发酵液的抗瘤作用可能是通过机体免疫系统介导的。     

软骨多糖诱导小鼠S180细胞凋亡的作用机制

目的研究软骨多糖对S180荷瘤小鼠的作用,并探讨其抑瘤作用机制。方法采用小鼠肉瘤S180细胞建立动物腹水瘤模型,通过腹腔注射软骨多糖进行治疗,治疗期间抽取腹水瘤细胞进行细胞生物学分析。通过HE染色,流式细胞术、TUNEL法检测细胞形态学方面、细胞周期及凋亡率的变化情况;通过免疫荧光方法检测Fas、增殖细胞核抗原(PCNA)的表达情况。结果软骨多糖可以明显提高S180荷瘤小鼠的生存率,细胞形态学观察可见细胞出现细胞质浓缩、核固缩及凋亡小体等现象。软骨多糖作用后的S180细胞,其细胞周期被阻遏于G2/M期,Fas蛋白的表达水平于给药24 h后升高,增殖细胞核抗原PCNA表达下降。结论软骨多糖可能通过影响肿瘤细胞周期和Fas、PCNA蛋白的表达来诱导S180细胞凋亡,并显著抑制肿瘤细胞的生长,延长S180荷瘤小鼠的生存时间,研究证实动物软骨多糖具有潜在的药用价值。     

中药麝香胶囊抑瘤作用的实验研究

目的探讨中药麝香胶囊对小鼠实验性肿瘤的疗效。方法昆明小鼠分别接种艾氏腹水癌、S-180、肝癌细胞株（hepatocellular carcinoma,HCC）三种癌细胞株,接种24h后开始给予中药麝香胶囊,每日1次,共10d。分高、中、低三个剂量给药（4、2、1g／kg以主药麝香药量计）。阳性对照以天仙丸胶囊（1g／kg）一次性灌服。阴性对照组以同体积生理盐水灌服。接种实体瘤动物于第10天处死,称瘤重,计算抑瘤率;接种腹水瘤动物,观察存活时间,计算生命延长率。结果中药麝香胶囊对实体癌有一定的抑瘤作用,但未达到药典规定的抑瘤率30％的要求;对腹水癌也有一定的抑瘤作用,但也未达到药典规定生命延长率50％的要求;统计学分析（P〉0．05）差异没有显著性。结论中药麝香胶囊抑瘤作用不明显,其配方及剂型有待进一步研究与改进。     

Gonadotropin-releasing hormone analog structural determinants of selectivity for inhibition of cell growth: support for the concept of ligand-induced selective signaling

GnRH and its receptor are expressed in human reproductive tract cancers, and direct antiproliferative effects of GnRH analogs have been demonstrated in cancer cell lines. The intracellular signaling responsible for this effect differs from that mediating pituitary gonadotropin secretion. The GnRH structure-activity relationship is different for the two effects. Here we report a structure-activity relationship study of GnRH agonist antiproliferative action in model cell systems of rat and human GnRH receptors stably expressed in HEK293 cells. GnRH II was more potent than GnRH I in inhibiting cell growth in the cell lines. In contrast, GnRH I was more potent than GnRH II in stimulating inositol phosphate production, the signaling pathway in gonadotropes. The different residues in GnRH II (His(5), Trp(7), Tyr(8)) were introduced singly or in pairs into GnRH I. Tyr(5) replacement by His(5) produced the highest increase in the antiproliferative potency of GnRH I. Tyr(8) substitution of Arg(8) produced the most selective analog, with very poor inositol phosphate generation but high antiproliferative potency. In nude mice bearing tumors of the HEK293 cell line, GnRH II and an antagonist administration was ineffective in inhibiting tumor growth, but D-amino acid stabilized analogs (D-Lys(6) and D-Arg(6)) ablated tumor growth. Docking of GnRH I and GnRH II to the human GnRH receptor molecular model revealed that Arg(8) of GnRH I makes contact with Asp(302), whereas Tyr(8) of GnRH II appears to make different contacts, suggesting these residues stabilize different receptor conformations mediating differential intracellular signaling and effects on gonadotropin and cell growth. These findings provide the basis for the development of selective GnRH analog cancer therapeutics that directly target tumor cells or inhibit pituitary gonadotropins or do both.     

Primaquine homodimers as potential antiplasmodial and anticancer agents

Primaquine homodimers, e.g. symmetric PQ-diamides of dicarboxylic acids containing 4 to 8 carbon atoms, were evaluated against Plasmodium berghei hepatic stages and P. falciparum blood stages, as well as against three cancer cell lines. Novel PQ-homodimers exerted much higher activity against hepatic stages, but less pronounced activity against blood stages in comparison to the parent drug. The submicromolar activity of succinic, fumaric and maleic derivatives against P. berghei was determined (IC₅₀ values: 726.2, 198.1 and 358.4 nM, respectively). Our results indicated that the length and type of spacer between two PQ moieties highly modified the antiproliferative activities of PQ-homodimers. The general antiproliferative activity of the adipic and mesaconic derivatives against three cancer cell lines (MCF-7, HCT116, H 460) was observed (GI₅₀ = 1.78–13.7 and 2.36–4.31 µM, respectively), but adipic derivative was less toxic to human embryonic kidney cells (HEK 293). High selectivity of fumaric and suberic derivatives against breast adenocarcinoma cell line MCF-7 was detected. These two compounds have shown no antiproliferative activity against other tumor cells and HEK 293.     

香菇C_（91－3）菌发酵液小鼠体内外抗肿瘤作用的研究

香菇Ｃ_（９１－３）菌是我们多年筛选出的一株经生物发酵后,其发酵液具有明显的抗肿瘤、抗细菌作用的真菌。研究结果表明：该发酵液具有明显体外抗肿瘤作用的活性物质。对ＭＨ_（１３４）、Ｘ５５５３、Ｃａ７６１／Ｌ、ＹＡＣ－１、Ｈ_（２２）、Ｋ５６２抑瘤率为７６．７～１００％。在体内抗肿瘤实验中,采用Ｈ_（２２）、Ｓ１８０腹水瘤的研究中,小鼠存活率分别为４０％、４５％。本文还对香菇Ｃ_（９１－３）菌发酵液抗肿瘤的机理进行了探讨。     

Ascitic and solid Ehrlich tumor inhibition by Chenopodium ambrosioides L. treatment

The leaves of Chenopodium ambrosioides L. [Chenopodiaceae] ('mastruz') have been indicated for the treatment of several diseases, among which the cancer. There are no results focusing the effect of C. ambrosioides treatment on tumor development in vivo. The aim of this study was to investigate the effect of treatment with C. ambrosioides on Ehrlich tumor development. Swiss mice were treated by intraperitoneal route (i.p.) with hydroalcoholic extract from leaves of C. ambrosioides (5 mg/kg) or with PBS (control group) 48 h before or 48 h later the Ehrlich tumor implantation. The tumor cells were implanted on the left footpad (solid tumor) or in the peritoneal cavity (ascitic tumor). To determine the solid tumor growth, footpad was measured each 2 days until the fourteenth day, when the feet were weighed. Ascitic tumor development was evaluated after 8 days of tumor implantation by quantification of the ascitic fluid volume and tumor cell number. The i.p. administration of C. ambrosioides extract before or after the tumor implantation significantly inhibited the solid and ascitic Ehrlich tumor forms. This inhibition was observed in ascitic tumor cell number, in the ascitic volume, in the tumor-bearing foot size and foot weight when compared to control mice. The treatments also increased the survival of tumor-bearing mice. In conclusion, C. ambrosioides has a potent anti-tumoral effect which was evident with a small dose and even when the treatment was given two days after the tumor implantation. This effect is probably related with anti-oxidant properties of C. ambrosioides.     

Up-regulation of vascular endothelial growth factor-D expression in clear cell renal cell carcinoma by CD74: a critical role in cancer cell tumorigenesis

Elevation of CD74 is associated with a number of human cancers, including clear cell renal cell carcinoma (ccRCC). To understand the role of CD74 in the oncogenic process of ccRCC, we ectopically expressed CD74 in human embryonic kidney 293 cells (HEK/CD74) and evaluated its oncogenic potential. Through overexpression of CD74 in HEK293 and Caki-2 cells and down-regulation of CD74 in Caki-1 cells, we show that vascular endothelial growth factor-D (VEGF-D) expression is modified accordingly. A significant, positive correlation between CD74 and VEGF-D is found in human ccRCC tissues (Pearson's correlation, r = 0.65, p < 0.001). In HEK/CD74 xenograft mice, CD74 significantly induced the formation of tumor masses, increased tumor-induced angiogenesis, and promoted cancer cell metastasis. Blockage of VEGF-D expression by small interference RNA resulted in a decrease in cell proliferation, invasion, and cancer cell-induced HUVEC migration enhanced by CD74. Furthermore, we provide evidence that the intracellular signaling cascade responsible for VEGF-D up-regulation by CD74 is both PI3K/AKT- and MEK/ERK-dependent, both of which are associated with NF-kappaB nuclear translocation and DNA-binding activity. These results suggest that VEGF-D is crucial for CD74-induced human renal carcinoma cancer cell tumorigenesis.     

Tumoricidal effect of human macrophage-colony-stimulating factor against human-ovarian-carcinoma-bearing athymic mice and its therapeutic effect when combined with cisplatin

The effect of human macrophage-colony-stimulating factor (hM-CSF) on tumoricidal activity was examined in athymic mice bearing the human ovarian cancer cell line, HRA, injected intraperitoneally (i.p.). The survival period and survival rate in the groups treated daily with hM-CSF were significantly longer (P<0.01) than in the untreated group. The peritoneal cell smears showed that ascitic tumor cells were markedly decreased in the hM-CSF-treated groups, and macrophages phagocytosed tumor cells, indicating a contact-mediated direct cytolysis. The combined therapeutic effects of cisplatin and hM-CSF on HRA-bearing athymic mice were also studied. The mean survival period was 25.4, 47.2, 42.4 and 67.4 days, respectively, in the untreated group, and in the groups treated with cisplatin alone, with hM-CSF alone, and with combined cisplatin and hM-CSF. The survival period and rate were significantly longer (P<0.01) in the group treated with combined cisplatin and hM-CSF than in those treated with cisplatin or hM-CSF alone, indicating the therapeutic effectiveness of the combined use. Morever, hM-CSF is effective against granulocytopenia due to bone marrow suppression caused by cisplatin. Our data demonstrate that hM-CSF administered i.p. has a tumoricidal activity in athymic mice bearing human ovarian cancer i.p., which is mediated by activated macrophages, and that the combined administration of cisplatin and hM-CSF has a significant therapeutic effect.     

蚯蚓中抗肿瘤蛋白组分的提取分离及其抗肿瘤活性

以赤子爱胜蚓为原料 ,通过蛋白质的丙酮沉淀和凝胶过滤 ,分离得到一组抗肿瘤活性蛋白成分 (简称蚯蚓抽提物 ) ;这组成分富含Fe、Zn、Cu以及Se等微量元素 ,蛋白含量为 6 0 4 3± 2 36 %(n =5 ) .体外实验中 ,通过MTT法和SRB法测定了蚯蚓抽提物对多种人癌细胞株 (HCT 116、SY5Y、K5 6 2、MGc80 3和HeLa)以及正常细胞株 (HEK2 93和COS 7)的抑制杀伤率 .结果表明 ,蚯蚓抽提物对癌细胞的杀伤有一定的选择性 ,受试人癌细胞达到 5 0 %生长抑制率所需作用浓度约 6 0～110mg L ;但是 ,10 0℃煮沸 5min后 ,该抑制活性完全消失 .通过纤维蛋白平板法 ,测得蚯蚓抽提物同时具有纤溶酶和纤溶酶原激活酶的活性 .体外测得丝氨酸蛋白酶抑制剂aprotinin和PMSF能显著抑制蚯蚓抽提物的细胞杀伤活性 .体内实验中 ,蚯蚓抽提物能有效延长荷瘤 (S180 )小鼠的生存时间 ,并使其身体机能得到明显改善 .腹膜内注射剂量为 2 8mg kg和 36mg kg时 ,生命延长率分别为135 3%和 12 3 5 % ,和标准药物 (环磷酰胺 )治疗后结果 (76 5 % )相比 ,有显著性差异     

Effect of saffron on thymocyte proliferation, intracellular glutathione levels and its antitumor activity.

Liposome encapsulation of saffron effectively enhanced its antitumor activity towards Sarcoma-180 (S-180) and Ehrlich ascites carcinoma solid tumors in mice. Significant inhibition (P < 0.001) in the growth of these tumors was observed as compared with vehicle (control) mice. In the presence of phytohemagglutinin (PHA), a T cell mitogen, saffron stimulated non-specific proliferation of lymphocytes in vitro. The intracellular reduced glutathione and related enzymes, i.e. glutathione reductase and glutathione-S-transferase, of S-180 tumor cells were significantly elevated when incubated with saffron, possibly acting to maintain functional levels of other antioxidants. Our studies indicate the antioxidant activity of saffron.