The anti-tumoral drug enzastaurin inhibits natural killer cell cytotoxicity via activation of glycogen synthase kinase-3β

Enzastaurin is a selective protein kinase Cβ inhibitor which is shown to have direct antitumor effect as well as suppress glycogen synthase kinase-3β (GSK-3β) phosphorylation (resulting in its activation) in both tumor tissues and peripheral blood mononuclear cells (PBMC). It is currently used in phase II trials for the treatment of colon cancer, refractory glioblastoma and diffuse large B cell lymphoma. In this study, the direct effect of enzastaurin on effector function of human natural killer (NK) cells was investigated. The results obtained showed that enzastaurin suppressed both natural and antibody-dependent cellular cytotoxicity (ADCC) of NK cells against different tumor targets. This inhibition was associated with a specific down-regulation of surface expression of NK cell activating receptor NKG2D and CD16 involved in natural cytotoxicity and ADCC respectively, as well as the inhibition of perforin release. Analysis of signal transduction revealed that enzastaurin activated GSK-3β by inhibition of GSK-3β phosphorylation. Treatment of NK cells with GSK-3β-specific inhibitor TDZD-8 prevented enzastaurin-induced inhibition of NK cell cytotoxicity. Apart from the known antitumor and antiangiogenic effects, these results demonstrate that enzastaurin suppresses NK cell activity and may therefore interfere with NK cell-mediated tumor control in enzastaurin-treated cancer patients.     

Immunomodulatory effect of polysaccharides from submerged cultured Cordyceps gunnii

Context: The genus Cordyceps (Clavicipitaceae) is a group of entomopathogenic fungi that is widely used as tonic food or invigorant with broad-spectrum medicinal properties in China. Cordyceps gunnii (Berk.)Berk (C. gunnii), is also well known as the Chinese rare caterpillar fungus and has similar pharmacological activities with Cordyceps sinensis (C. sinensis). Polysaccharides (PS) from various Cordyceps species have demonstrated many interesting biological activities, including antitumor, immunopotentiation, hypoglycemic, and hypocholesterolemic activities. Objective: To investigate the effect of C. gunnii PS on the immunostimulatory antitumor function and expression of immune related cytokines in normal, immuno-suppressive, and H22-bearing mice, respectively. Methods: C. gunnii PS were extracted with hot water at 80°C for 2 h. Normal, immuno-suppressive, and H22-bearing mice were treated with PS respectively. By detecting the value of macrophage phagocytic index, proliferation of lymphocytes, natural killer (NK) cell activity and expression of related cytokines, interleukin (IL-4), tumor necrosis factor-α (TNF-a) and interferon-γ (IFN-γ), and tumor inhibition index in H22-bearing mice additionally, the effect of PS on immunostimulatory antitumor function and its mechanism were studied. Results: The total sugar content of the PS was determined to be 95% after purification. PS markedly increased the thymus and spleen indexes, the macrophage phagocytosis, the proliferation of splenic cells, and the level of IFN-γ and TNF-α. In tumor growth inhibition test, PS showed remarkable inhibition effects. Conclusion: PS from the C. gunnii could enhance nonspecific immunological function, humoral immunity, cellular immunity in mice, and inhibit tumor growth.     

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。     

New anticancer agents: in vitro and in vivo evaluation of the antitumor and antimetastatic actions of various compounds isolated from medicinal plants

In this review, in the search for the development of new anticancer drugs, the effects of compounds isolated from various medicinal plants on tumor growth and metastasis, using mice bearing a highly metastatic drug-resistant mouse tumor, were studied. The antitumor and antimetastatic actions of stilbene derivatives isolated from Polygonum and Cassia species were examined. Among the stilbene derivatives, resveratrol and cassiagrol A (stilbene dimer) displayed antitumor and antimetastatic actions through the inhibition of tumor-induced neovascularization in in vitro and in vivo models. It was found that two chalcone derivatives from Angelica keiskei roots also inhibited tumor growth and metastasis in tumor-bearing mice through the inhibition of tumor-induced neovascularization and/or the inhibition of immune suppression caused by tumors. Recently, basidiomycete fungi have been used for the treatment of cancer. Then, the low molecular weight substances were isolated from Agaricus blazei and Ganoderma lucidum as antitumor and antimetastatic substances. It is suggested that these substances of basidiomycete also inhibited tumor growth and metastasis to the lung through the inhibition of tumor-induced neovascularization and/or the inhibition of immune suppression caused by tumors.     

关于抗肿瘤药物的研究 Ⅲ.利用体外培养吉田腹水肉瘤篩选抗肿瘤药物

本文介紹利用短期体外培养吉田腹水肉瘤細胞进行药物篩选的方法.所用培养基为馬血清平衡盐溶液,每毫升接种細胞数为6—10万,于加入药物后24-48小时进行細胞計数,計算药物对細胞生长的抑制百分率,并以形态观察作为輔助指标. 曾进行5种已知抗肿瘤药的敏感試驗,发現药物的作用强度为:氮芥>氧化氮芥>新恩必恨>丁二醇二甲磺酸酯(myleran)>6-巯基嘌呤.此外,过篩了24种合成药和93种天然药物成分,其中有作用者分別为41.7%和38.7%.将篩选結果与动物瘤体內結果作比較,发現二者间有一定的相关.最后对方法的优缺点进行了討論.     

Immunological studies on sporamycin-treated animals

Sporamycin showed a remarkable tumor regressive activity against sarcoma-180 with a single 5 mg/kg dose of intravenous administration. This antitumor effect on tumor and host animals was examined immunologically. As the results: (1) When sarcoma-180 tumor cells were used as an antigen macrophage migration inhibition reaction by spleen cells derived from the tumor-bearing mice treated with sporamycin was positive at day 7 approximately 14 after the medication and was negative thereafter. (2) The delayed hypersensitivity tested by the foot-pad reaction was positive in tumor-bearing mice treated with sporamycin, and no decrease of foot pad reaction was observed, whereas this reaction decreased remarkably in non-treated tumor-bearing mice. (3) Sarcoma-180 tumor cells were mixed with spleen cells derived from sporamycin-treated mice, and were inoculated into normal dd mice. The growth of tumor cells was inhibited markedly, but no inhibition of tumor growth was observed in case of spleen cells derived from non-treated tumor bearing mice. (4) Combined treatment of sporamycin with PS-K, an immunopotentiator, showed a remarkable synergistic effect.     

Immunomodulatory effect of polysaccharides from submerged cultured Cordyceps gunnii

Context: The genus Cordyceps (Clavicipitaceae) is a group of entomopathogenic fungi that is widely used as tonic food or invigorant with broad-spectrum medicinal properties in China. Cordyceps gunnii (Berk.)Berk (C. gunnii), is also well known as the Chinese rare caterpillar fungus and has similar pharmacological activities with Cordyceps sinensis (C. sinensis). Polysaccharides (PS) from various Cordyceps species have demonstrated many interesting biological activities, including antitumor, immunopotentiation, hypoglycemic, and hypocholesterolemic activities.

Objective: To investigate the effect of C. gunnii PS on the immunostimulatory antitumor function and expression of immune related cytokines in normal, immuno-suppressive, and H22-bearing mice, respectively. Methods: C. gunnii PS were extracted with hot water at 80°C for 2 h. Normal, immuno-suppressive, and H22-bearing mice were treated with PS respectively. By detecting the value of macrophage phagocytic index, proliferation of lymphocytes, natural killer (NK) cell activity and expression of related cytokines, interleukin (IL-4), tumor necrosis factor-α (TNF-a) and interferon-γ (IFN-γ), and tumor inhibition index in H22-bearing mice additionally, the effect of PS on immunostimulatory antitumor function and its mechanism were studied.

Results: The total sugar content of the PS was determined to be 95% after purification. PS markedly increased the thymus and spleen indexes, the macrophage phagocytosis, the proliferation of splenic cells, and the level of IFN-γ and TNF-α. In tumor growth inhibition test, PS showed remarkable inhibition effects.

Conclusion: PS from the C. gunnii could enhance nonspecific immunological function, humoral immunity, cellular immunity in mice, and inhibit tumor growth.     

蜈蚣提取物体外抗肿瘤活性比较及体内活性评价

目的：比较蜈蚣干品和鲜品不同提取物的体外抗肿瘤活性，并评价其体内抗肿瘤活性。方法：蜈蚣干品和鲜品的水、30%、50%、70%及95%乙醇提取物分别处理MGC803细胞，采用MTT法检测细胞生长抑制率，计算半数抑制浓度（IC₅₀）；观察蜈蚣提取物对H22腹水瘤小鼠的生存期和实体瘤小鼠的抑瘤率的影响。结果：蜈蚣干品和鲜品不同提取物对肿瘤细胞的IC₅₀值相近，而蜈蚣水提取物的IC₅₀值相对最小。蜈蚣水提取物能延长腹水瘤小鼠的生存期；对实体瘤小鼠瘤重有明显的抑制作用，但中剂量和高剂量组有部分动物加速死亡。结论：多种提取方法得到的蜈蚣提取物均具有体外抗肿瘤活性，蜈蚣水提取物有一定的体内抗肿瘤活性，但超剂量使用有一定的毒性。     

Synthesis of polyrotaxane-camptothecin conjugates and evaluation of its anti-tumor effect

To prepare polyrotaxane-camptothecin conjugates and evaluate its anti-tumor effect, polyrotaxane-camptothecin conjugates were successfully synthesized, and the release behavior was performed; MTT assay and cell morphology were used to examine the inhibition of cells' proliferation effect in vitro. The experimental study of the antitumor effect on S180 mice in vivo was also performed to further evaluate the anti-tumor effect of conjugate. The result showed polyrotaxane-camptothecin conjugates can effectively inhibit the proliferation in a dose dependent effect. In vivo study and cell morphology observation of S180 mice showed significant decrease in growth of tumor, degree of tumor infiltration and blood vessel number. The result indicated anti-tumor mechanism may be through affect the angiogenesis and reduced blood supply to tumor cells and then leading to necrosis.     

灵芝孢子粉对荷瘤小鼠树突状细胞的影响及其抗瘤效应

目的通过研究灵芝孢子粉对荷H22肝癌小鼠髓源性树突状细胞(DC)的影响及其抗肿瘤效应,探讨其抗肿瘤机制。方法应用不同浓度灵芝孢子粉(1、2、3g·kg-1·d-1)连续14d灌胃荷瘤小鼠后,处死小鼠,无菌获取骨髓细胞,粒细胞巨噬细胞刺激因子(GMCSF)和白细胞介素(IL)4体外诱导骨髓细胞生成DC,同时培养液中继续加入不同浓度灵芝孢子粉(0.8、3.2、12.8mg/L)。通过显微镜、流式细胞仪检测技术及细胞毒性实验观察不同浓度灵芝孢子粉对DC的影响,并通过脾指数、胸腺指数及肿瘤抑制率来观察其抗肿瘤效应。结果灵芝孢子粉能够促进荷瘤小鼠髓源性DC的分化、成熟及表面分子CD11a、CD86的表达,增强DC诱导的细胞毒性T淋巴细胞反应;灵芝孢子粉能够使荷瘤小鼠免疫器官脾脏和胸腺的重量增加,并明显抑制肿瘤的生长,存在量效关系。结论本实验表明灵芝孢子粉能够通过刺激DC的分化成熟,提高其抗原提呈能力,进而起到提高机体细胞免疫,抑制肿瘤生长的作用。     

凡德他尼衍生物的抗肿瘤活性筛选研究

目的对合成的一系列凡德他尼衍生物进行体内外抗肿瘤活性的筛选,为寻找低毒高效的新型酪氨酸激酶抑制剂研究提供依据。方法体外筛选采用均相时间分辨荧光(HTRF)法和磺酰罗丹明B(SRB)法分别进行激酶和细胞的筛选;采用经典的急性毒性实验方法,并建立移植人非小细胞肺癌H1975裸鼠模型评价其抗肿瘤活性。结果 HTRF结果显示有6个活性较好的化合物(TY8115、TY8119、TY8122、TY8128、TY8129、TY8131),其中TY8115对VEGFR-2和EGFR抑制作用均好于凡德他尼;SRB结果显示这些活性化合物对选用的3种靶细胞(A431、H1975、A549)均有不同程度的抑制作用,其中TY8115的肿瘤细胞增殖抑制作用最明显,且对非靶细胞(MDA-MB-231)生长影响很小;急性毒性实验结果显示TY8115没有表现出毒性反应;体内抗肿瘤活性研究结果显示TY8115对肺癌H1975具有疗效,75、150 mg/kg TY8115对H1975的相对肿瘤增殖率分别为54.44%、39.54%。结论化合物TY8115具有良好的抗肿瘤活性,并且毒副作用小,具有发展成为一种新型酪氨酸激酶抑制剂的潜力。     

SEA体内杀伤S180肉瘤效果及其细胞因子的测定

目的:研究SEA抗肿瘤的效果及其对细胞因子干扰素(IFN)-γ的影响,探讨其作用机制。方法:采用S180细胞建立小鼠肉瘤模型40只进行体内抑瘤实验,随机分成4组,包括SEA高剂量组(n=10)、SEA中剂量组(n=10)、SEA低剂量组(n=10)和对照组(n=10),前3组分别注射0.5、0.25、0.05mg/kg的SEA,对照组注射生理盐水,接种第5天起测量各组小鼠肿瘤体积,绘制肿瘤生长曲线并计算抑瘤率。治疗结束后取小鼠血清并测定IFN-γ的含量。结果:SEA高、中和低剂量组抑瘤率分别为(44.00±3.21)%、(3.92±2.12)%和(2.33±1.95)%,SEA高剂量组脾脏指数和IFN-γ的含量明显增加,与对照组比较差异均有统计学意义(P<0.01),但肿瘤质量却明显减小。结论:SEA通过激活T细胞,分泌细胞因子IFN-γ抑制小鼠体内S180肉瘤生长。     

In vitro and in vivo antitumor effect of 5-FU combined with piplartine and piperine

It has been reported that piplartine and piperine, alkaloid/amide compounds from Piper species, show antitumor activities. In the present paper, the effects of the combination of 5-fluorouracil (5-FU) with piplartine or piperine was determined using in vitro and in vivo experimental models. Hematological and biochemical analyses, as well as histopathological and morphological analyses of the tumor and the organs, including liver, spleen and kidney, were performed in order to evaluate the toxicological aspects associated with different treatments. The incubation of tumor cell lines with 5-FU in the presence of piplartine or piperine produced an increase in growth inhibition, as observed by lower IC50 values for 5-FU. These effects were also observed in vivo, where the combination with piplartine but not piperine with 5-FU led to a higher tumor growth inhibition. The results indicated that either piplartine- or 5-FU-treated animals showed a low inhibition rate when they were used individually at low doses of 28.67% and 47.71%, respectively, but when they were combined at the same dose, the inhibition rate increased significantly to 68.04%. The histopathological analysis showed that the livers and the kidneys of treated animals were only slightly and reversibly affected. Neither the enzymatic activity of transaminases nor the urea levels were significantly modified when compared with the control group. Hematological analysis showed leukopenia after 5-FU treatment, which was reversed by the combined use of piplartine and piperine. These findings indicate that piplartine may enhance the therapeutic effectiveness of chemotherapeutic drugs, and moreover, this combination could improve immunocompetence hampered by 5-FU.     

<Emphasis Type="Italic">Pulsatilla</Emphasis> saponin D: The antitumor principle from<Emphasis Type="Italic">Pulsatilla koreana</Emphasis>

By bioassay-guided separation, an already known saponin, Pulsatilla saponin D was isolated from the root of Pulsatilla koreana Nakai as a antitumor component when evaluated by in vivo antitumor activity as well as in vitro cytotoxic activity test. It showed potent inhibition rate of tumor growth (IR, 82%) at the dose of 6.4 mg/kg on the BDF1 mice bearing LLC cells.     

Characteristics of tissue distribution of various polysaccharides as drug carriers: influences of molecular weight and anionic charge on tumor targeting

Using the Walker 256 model for carcinosarcoma-bearing rats, we intravenously administered 5 polysaccharide carriers with various molecular weights (MWs) and electric charges and tested for their plasma and tissue distribution. Two carriers, carboxymethylated-D-manno-D-glucan (CMMG) and CMdextran (CMDex), showed higher plasma AUC than the other carriers tested, namely, CMchitin (CMCh), N-desulfated N-acetylated heparin (DSH), and hyaluronic acid (HA). This was consistently found to be true over the range of MWs tested. For CMDex, the maximum value of plasma AUC was obtained when the MW exceeded 150 kDa. As for the anionic charge, CMDex (110-180 kDa) with a degree of substitution (DS) of the CM groups ranging from 0.2 to 0.6, showed maximum plasma AUC values. Twenty-four hours after administration, the concentration of CMDex (180-250 kDa; DS: 0.6-1.2) in tumors was more than 3% of dose/g--approximately 10-fold higher than those observed with CMCh, DSH and HA. Doxorubicin (DXR) was bound to these carriers via a peptide spacer, GlyGlyPheGly (GGFG), to give carrier-GGFG-DXR conjugates (DXR content: 4.2-7.0 (w/w)%), and the antitumor effects of these conjugates were tested with Walker 256 carcinosarcoma-bearing rats by monitoring the tumor weights after a single intravenous injection. Compared with free DXR, CMDex-GGFG-DXR and CMMG-GGFG-DXR conjugates significantly suppressed tumor growth, while the CMCh-GGFG-DXR, DSH-GGFG-DXR, and HA-GGFG-DXR conjugates in a similar comparison showed weak tumor growth inhibition. These findings suggest that the antitumor effect of the carrier-DXR conjugates was related to the extent with which the carriers accumulated in the tumors.     

化合物209对人结直肠癌细胞HT-29的体内和体外作用(英文)

化合物209是一个新合成的氨基二硫代甲酸酯类化合物,它在体外水平可以抑制肿瘤细胞的增殖,但是化合物209的体内抗肿瘤作用及其抗肿瘤机制并不明确。本文探究了化合物209对人结直肠癌细胞HT-29的作用并初步探讨了相关机制。体外研究表明,化合物209可以显著抑制HT-29细胞的增殖;体内研究结果表明,化合物209可以显著抑制裸鼠HT-29移植瘤的生长,但是对裸鼠体重和白细胞无影响。流式细胞分析实验结果表明,化合物209可将HT-29细胞阻滞于细胞周期的G1期。同时,化合物209能上调体外培养HT-29细胞中p27,cyclin E,CDK2,cyclin D1和CDK4的表达。在体内瘤组织中上述蛋白表达情况与体外实验结果一致。这些结果说明,化合物209具有较好的抗肿瘤活性,其抗肿瘤作用与细胞周期阻滞及其相关蛋白的表达变化有关。     

人参皂苷Ro对胃癌细胞MFC增殖和凋亡的影响

目的 探讨人参皂苷Ro对胃癌细胞MFC增殖和凋亡的影响.方法 采用MTT染色法检测MFC细胞的增殖抑制率;流式细胞仪检测MFC细胞周期和凋亡情况;MFC移植瘤小鼠,经受试药物处理后,剥取瘤组织称量并计算抑瘤率.结果 与对照组比较,人参皂苷Ro在6～96 μg· mL-1范围内对MFC细胞增殖的抑制率随浓度的增加而显著升高.与对照组比较,人参皂苷Ro高剂量组对MFC细胞G0/G1期和凋亡率明显升高,G2/M期,S期明显降低;人参皂苷Ro各剂量组均能显著抑制肿瘤生长,与模型组比较,各给药组肿瘤的质量明显降低,其中,高剂量组的抑瘤率为62.7％.结论 人参皂苷Ro对MFC移植瘤具有抑制生长的作用,这可能与诱导MFC细胞凋亡和抑制增殖有关.     

The effect of alkaloid from Oxytropis ochrocephala on growth inhibition and expression of PCNA and p53 in mice bearing H22 Hepatocellular Carcinoma

To study the antitumor activity of alkaloid extracted from Oxytropis ochrocephala and its possible mechanism, we observed the effect of alkaloid on tumor weight and expression of PCNA and p53 in mice bearing H(22) hepatocellular carcinoma by means of immunohistochemistry SP method. After treatment with alkaloid from Oxytropis ochrocephala, the results showed that alkaloid administration (25 and 50 mg/kg body weight, p.o.) could inhibit H(22) hepatocellular carcinoma growth to various extent, and the rates of inhibition were 48.5% and 57.7% respectively (p<0.01). The antitumor activity of the alkaloid is in a dose dependent manner, with no signs of toxicity to weight, kidney and liver. The sections of tumor showed the number of tumor cell decreased and nucleus appeared putrescence such as nucleus atrophy, disintegrating and dissolving. Meanwhile, the expression of PCNA and mutant p53 protein positive cell numbers in mice bearing H(22) hepatocellular carcinoma also suppressed by alkaloid (p<0.05). It suggested that Alkaloid from Oxytropis ochrocephala showed antitumor effect and its possible mechanism might be associated with the expression inhibition of PCNA and mutant p53 protein. Further studies are needed to explore the antitumor activity of the other compounds of Oxytropis ochrocephala and to specify their possible mechanism of action.     

Mechanisms of potentiation of antitumor activity of 5-fluoro-2'-deoxyuridine in the adenocarcinoma 755 system by guanosine 5'-monophosphate

The effect of guanosine 5'-monophosphate (GMP) on the antitumor activity of 5-fluoro-2'-deoxyuridine (FdUrd) was investigated by using a solid tumor, adenocarcinoma 755. FdUrd only slightly inhibited the tumor growth even at the maximum tolerated dos (ILS,6%). GMP at 300 mg/kg/day markedly potentiated the inhibition of tumor growth by FdUrd (ILS, 61%). When tumor-bearing mice were treated with the combination of 3H-FdUrd and GMP, 3H-FdUrd was significantly incorporated into tumor RNA as compared to the mice not given GMP, although the 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP) level in the tumor in the combination with GMP was decreased. The increased incorporation of 3H-FdUrd into RNA ([FUra]RNA) of the tumor was due to an increased level of FUra in plasma and tumor. On the other hand, incorporation of 3H-FdUrd into the RNA of the small intestine, which is one of main target tissues of FUra toxicity, was not increased. Thus, the potentiating effect of GMP on the antitumor activity of FdUrd is apparently due to the specifically increased FdUrd and FUra levels in the tumor. The combination with GMP caused marked incorporation of 3H-FdUrd into RNA in the tumor but uptake of 3H-FdUrd into RNA in the small intestine was not increased. These results suggest that GMP can potentiate the antitumor activity of FdUrd without increasing gastro-intestinal toxicity.     

d-宁烯、丹参及姜黄素衍生物对ras基因产物膜结合和细胞间隙信息传导的影响

目的旨在寻找新型抗肿瘤药物,进一步研究d-宁烯、丹参及姜黄素衍生物的抗肿瘤机理。采用分子生物学方法及划痕标记染料示踪技术,研究了4种人实体瘤起源的细胞系的细胞间隙信息传导(GJIC)、H-ras癌基因表达以及ras癌基因产物(P21^ras蛋白)表达状态,并观察了4种天然产物对它们的影响。结果表明,细胞内染料传输功能的丧失与ras基因突变率呈正相关;单萜化合物d-宁烯和酚类化合物丹参衍生物的抗肿瘤作用可能与抑制P21^ras蛋白膜结合和增强细胞间隙信息传导功能有关。提示Ras癌基因产物P21^ras蛋白膜结合的抑制与细胞间隙信息传导功能的增强有直接关系。