大黄、当归、木香的不同配伍抑瘤作用的药效学研究

目的 观察大黄.当归,木香的不同配伍组合对Lewis肺癌.肉瘤-s180、艾氏腹水癌荷瘤小鼠抑瘤作用的影响,探讨.大黄.当归、木香的最佳配伍关系.方法 制备lewis.肉瘤-s180、艾氏腹水癌荷瘤小鼠动物模型,随机分为模型组,大黄.当归组,大黄组,大黄、木香组,大黄.木香、当归组;通过测定荷瘤小鼠的瘤重.计算抑瘤卒观察不同配伍组对肿瘤生长的影响.结果 与模型组比较,大黄当归组对Lcewis肺癌、肉瘤-sI80.艾氏腹水癌抑瘤率分别达到78.52%、44.57%.64.95%;大黄.木香.当归组对Lewis ,肺癌.肉瘤-s180,艾氏腹水癌的抑瘤率分别达到4 7.53%、32.57%、52.99%,两个用药组合均具有很好的抑瘤作用(P＜0.05).     

芦荟多糖对小鼠抗肿瘤作用的实验研究

目的研究中华芦荟多糖(AP)对小鼠的抗肿瘤作用。方法观察不同浓度中华芦荟多糖对移植性S180肿瘤小鼠和EAC荷瘤小鼠的抑瘤率,对EAC荷瘤小鼠肿瘤坏死因子(TNF)含量的影响,并以脾系数为指标,观察中华芦荟多糖对环磷酰胺(CTX)的增效减毒作用。结果不同浓度的中华芦荟多糖对S180荷瘤小鼠和EAC荷瘤小鼠的肿瘤均具有不同程度的抑制作用。结论中华芦荟多糖可显著改善CTX引起的机体生理机能紊乱,提高其抑瘤作用。     

皖南红豆杉枝叶中总黄酮抗肿瘤活性研究

以MTT法考察皖南红豆杉枝叶中总黄酮的体外抗肿瘤活性;以小鼠移植瘤模型测定总黄酮对荷瘤小鼠抑瘤率和腹腔巨噬细胞吞噬能力,研究其体内抗肿瘤活性和免疫调节作用。结果表明,总黄酮对S180细胞生长有显著抑制作用,呈现剂量依赖关系。体内试验表明,与生理盐水组相比,总黄酮对小鼠S180实体瘤有显著抑制作用（P〈0.01）,可提高荷瘤小鼠的脾指数。与环磷酰胺组相比,总黄酮可使荷S180实体瘤小鼠腹腔巨噬细胞吞噬能力显著升高（P〈0.01）,提示其对小鼠无免疫抑制作用。     

透明质酸-阿魏酸靶向药物对Lewis肺癌荷瘤小鼠的抑制作用研究

目的:合成透明质酸-阿魏酸靶向药物并对其进行Lewis肺癌荷瘤小鼠的抑制作用研究。方法:使用丝氨酸甲酯作为中间连接物合成HA-FA靶向药物,采用紫外扫描图及1HNMR图谱进行结构确证;Bal b/c小鼠右腋皮下接种Lewis肺癌瘤株,连续腹腔注射给药20 d后,处死动物,摘取瘤体称重,并计算肿瘤生长抑制。结果:通过紫外扫描图可以初步证明HA-FA成功合成;1HNMR图谱进行结构确证。HA-FA对Lewis肺癌荷瘤小鼠的抑瘤率为57.97%(与模型组比较,P0.05)。结论:HA-FA对Lewis肺癌荷瘤小鼠肿瘤生长具有明显抑制作用。     

灵芝精粉与灵芝孢子粉对Lewis肺癌模型小鼠免疫功能的影响

目的研究灵芝精粉与灵芝孢子粉对Lewis肺癌模型小鼠免疫功能的影响,并比较二者在肿瘤治疗中的免疫调节作用。方法经C57BL/6小鼠右腋皮下接种1×106个Lewis肺癌细胞,建立小鼠Lewis肺癌模型。于接种肿瘤细胞次日至第20天,分别隔日灌胃灵芝孢子粉(D1)、灵芝精粉001(D2)及灵芝精粉002(D3),剂量均为0.5g/kg,同时设立对照组和肿瘤组(灌胃等体积生理盐水)。监测各组小鼠体重及瘤体积,第21日处死小鼠,称量瘤重,并无菌制备脾细胞悬液,检测小鼠NK细胞杀伤活性,流式细胞术检测小鼠脾细胞中CD4+、CD8+T细胞和B细胞数量,淋巴细胞转化试验检测脾细胞中T、B细胞增殖功能。结果与对照组比较,肿瘤组小鼠脾细胞中T、B细胞增殖功能、细胞数量及NK细胞杀伤活性均有不同程度的下降。与肿瘤组相比,3种灵芝制剂均可提高Lewis肺癌模型小鼠NK细胞的杀伤活性和T、B细胞增殖功能,其中,以D2作用最为明显(P<0.05);3种灵芝制剂均可不同程度地提高Lewis肺癌模型小鼠CD4+、CD8+T细胞数量,其中,以D3作用最为显著(P<0.05)。3种灵芝制剂对Lewis肺癌模型小鼠的体重、瘤重、瘤体积及B细胞数量均无明显影响。结论灵芝制剂对Lewis肺癌小鼠的免疫功能具有正向调节作用,其中灵芝精粉的作用优于灵芝孢子粉。     

海嘧啶对小白鼠S_(180)、EAC和H_(22)肿瘤细胞膜钠泵活性的影响

目的观察海嘧啶对小白鼠实体型肿瘤(S180)和腹水型肿瘤(EAC,H22)细胞膜钠泵活性的影响。方法荷瘤小鼠分为治疗组和对照组,分别测定各组瘤细胞膜钠泵活性。结果海嘧啶对小鼠S180、EAC及H22肿瘤细胞膜钠泵活性有明显的抑制作用。结论海嘧啶抗肿瘤的药理机制与其能够抑制肿瘤细胞膜钠泵活性有关。     

治疗用卡介苗抗肿瘤动物实验方法的建立

目的 建立治疗用卡介苗在小鼠体内抗肿瘤的药效学实验方法。方法 观察治疗用卡介苗(BCG)对小鼠实体瘤S180和腹水瘤(EC)的抑瘤作用。结果BCG可明显减轻S180荷瘤鼠的瘤重,其瘤重抑制率>30％;可明显延长EC腹水瘤小鼠的生存时间,其生命延长率>75％。其中以12.5％和6.25mg/kg两剂量组的效果较佳。结论 该方法可用于卡介苗抗肿瘤动物药学实验。     

重组腺病毒Ad-CMV-TK的构建及其对Lewis细胞的抑制作用

目的构建重组腺病毒Ad-CMV-TK,并检测Ad-CMV-TK/GCV系统对小鼠Lewis肺癌细胞的抑制作用。方法PCR扩增HSV-TK基因,以复制缺陷型腺病毒为载体构建重组腺病毒Ad-CMV-TK,在HEK293细胞中包装,CsCl2梯度离心法纯化后,感染Lewis肺癌细胞,Western blot检测HSV-TK蛋白的表达;MTT法检测Ad-CMV-TK/GCV系统对Lewis细胞的体外抑制作用;同时检测Ad-CMV-TK/GCV系统对C57荷瘤小鼠的抑瘤作用。结果重组腺病毒Ad-CMV-TK感染的Lewis细胞可特异地表达HSV-TK蛋白,Ad-CMV-TK/GCV系统在体外能明显抑制Lewis细胞的生长,当GCV浓度为100μmol/L时,Lewis细胞的存活率仅为16%;在体内能明显抑制荷瘤小鼠肿瘤的生长,与Ad-Blank/GCV对照组和PBS对照组相比,差异具有统计学意义。结论已成功构建了重组腺病毒Ad-CMV-TK,Ad-CMV-TK/GCV系统在体内及体外对小鼠Lewis肺癌细胞均有明显的抑制作用,为进一步研究其抗肿瘤机制及应用奠定了基础。     

氨氯地平对小鼠肝癌H_(22)细胞的抑制作用及其机制

目的探讨氨氯地平对小鼠肝癌H22细胞的抑制作用及其机制。方法采用MTT法检测氨氯地平对小鼠肝癌H22细胞增殖的影响。建立肝癌H22荷瘤小鼠模型,观察氨氯地平对荷瘤小鼠肿瘤的抑瘤率;HE染色观察肿瘤组织的病理改变;免疫组织化学方法检测肿瘤组织中bcl-2和bax蛋白的表达水平。结果氨氯地平作用48h可显著抑制小鼠肝癌H22细胞增殖,且呈剂量依赖性,其半数抑制浓度为5.6×10-3mg/ml。体内试验显示,氨氯地平(3和10mg/kg·d)灌胃给药10d能显著抑制H22荷瘤小鼠肿瘤生长;HE染色可见,氨氯地平给药组小鼠肿瘤细胞核染色变浅,细胞排列紧密;免疫组化显示,氨氯地平给药组小鼠肿瘤组织内bcl-2蛋白表达下调,而bax蛋白表达上调。结论氨氯地平具有明显的抑瘤作用,其抑瘤机制可能与诱导肿瘤细胞凋亡相关。     

抗瘤酮A_(10)、5Fu、GM-CSF和IL-2的抗癌效果

目的　比较A1 0 、5Fu、GM CSF和IL 2的抗癌效果。方法　选取NIH小鼠 ,分别腹腔接种肝腹水HeP A 2 2细胞、骨肉瘤S180细胞及胃癌MFC细胞 ,10d后将注射每种细胞的小鼠分为 3组 ,每组 12只 ,分别于小鼠腹腔注射A1 0 、5Fu、GM CSF和IL 2 ,每日 1次 ,每次 1ml 只 ,连续给药 8d后改为每 3d给药 1次 ,观察小鼠生存时间及抑瘤效果。结果　A1 0 、5Fu、GM CSF及IL 2均能延长小鼠生存时间 ,A1 0 抑制肝HeP A 2 2肿瘤效果显著。结论　A1 0 、5Fu、GM CSF和IL 2均有抑制癌细胞生长作用     

A Novel LMP1 Antibody Synergizes with Mitomycin C to Inhibit Nasopharyngeal Carcinoma Growth in Vivo Through Inducing Apoptosis and Downregulating Vascular Endothelial Growth Factor

Combined therapy emerges as an attractive strategy for cancer treatment. The aim of this study was to investigate the inhibitory effects of mitomycin C (MMC) combined with a novel antibody fragment (Fab) targeting latent membrane protein 1 (LMP1) on nasopharyngeal carcinoma (NPC) xenograft nude mice. The inhibitory rates of MMC (2 mg/kg), Fab (4 mg/kg), MMC (2 mg/kg) + Fab (4 mg/kg), and MMC (1 mg/kg) + Fab (4 mg/kg) were 20.1%, 7.3%, 42.5% and 40.5%, respectively. Flow cytometry analysis showed that the apoptotic rate of xenograft tumor cells in the MMC and Fab combination group was 28 ± 4.12%, significantly higher than the MMC (2 mg/kg) group (P < 0.01). Immunohistochemical staining showed that VEGF expression in NPC xenografts was significantly inhibited in the combination group compared to the Fab (4 mg/kg) group (P < 0.05). In conclusion, both MMC and Fab could inhibit NPC xenograft tumor growth in vivo and combination therapy showed apparent synergistic anti-tumor effects, which may be due to the induction of tumor cell apoptosis and the downregulation of VEGF expression. These results suggest that the novel combined therapy utilizing traditional chemotherapeutics and antibody-targeted therapy could be a promising strategy for the treatment of NPC.     

Combination of Low Concentration of (?)-Epigallocatechin Gallate (EGCG) and Curcumin Strongly Suppresses the Growth of Non-Small Cell Lung Cancer in Vitro and in Vivo through Causing Cell Cycle Arrest

(−)-Epigallocatechin gallate (EGCG) and curcumin are two naturally derived agents that have been widely investigated worldwide. They exhibit their anti-tumor effects in many types of cancers. In the current study, the effect of the combination of the two agents on non-small cell lung cancer (NSCLC) cells was investigated. The results revealed that at low concentrations, the combination of the EGCG and curcumin strongly enhanced cell cycle arrest. Flow cytometry analysis showed that the cells were arrested at G1 and S/G2 phases. Two main cell cycle related proteins cyclin D1 and cyclin B1 were significantly inhibited at the present of EGCG and curcumin. EdU (5-ethynyl-2′-deoxyuridine) fluorescence staining showed that the DNA replication was significantly blocked. A clonal growth assay also confirmed a marked repression of cell growth. In a lung cancer xenograft node mice model, combination of EGCG and curcumin exhibited protective effect against weight loss due to tumor burden. Tumor growth was strongly repressed by the combination of the two agents, without causing any serious side-effect. Overall, these results strongly suggest that EGCG in combination with curcumin could be a candidate for chemoprevention agent of NSCLC.     

Synthesis and Biological Evaluation of 10-Substituted Camptothecin Derivatives with Improved Water Solubility and Activity

Despite remarkable clinical achievements, camptothecin (CPT) still suffers from poor solubility and severe toxicity. Therefore, it is necessary to redevelop CPT derivatives as supplementary antitumor agents with good water solubility and small side effects. In this work, 27 camptothecin derivatives were synthesized and screened for their cytotoxicity against A549 (lung) and HCT-116 (colon) cancer cell lines. Among them, compound B7 , 7-ethyl-10-(2-oxo-2-(4-methylpiperidin-1-yl)ethoxy)camptothecin,was demonstrated in vitro to be a more potent antitumor agent than SN-38 by comparison of their inhibitory activities against cell proliferation and colony formation and interference effect on process of cell cycle and cell apoptosis. Additionally, a molecular docking model revealed that B7 can interact with the topoisomerase I–DNA complex, and that the solubility of B7 reached 5.73 μg/mL in water. Moreover, B7 significantly inhibited tumor growth in an A549 xenograft model at dosages of 0.4 and 2.0 mg/kg, and exhibited minimum lethal doses comparable to those of irinotecan. These results indicated that B7 , with improved solubility, enhanced activity and acceptable acute toxicity, can be used as a lead compound for the development of novel anticancer agents.     

Anti-Tumor Effect of Heat-Killed Bifidobacterium bifidum on Human Gastric Cancer through Akt-p53-Dependent Mitochondrial Apoptosis in Xenograft Models

Paraprobiotics, inactivated microbial cells, regulate immune system and exhibit antioxidant and anti-inflammatory activities in patients with weakened immunity or the elderly. This study evaluated the anti-tumor effects of heat-killed Bifidobacterium and Lactobacillus on human gastric cancer MKN1 cells in vitro and in vivo in xenograft animal models. First, cytotoxicity and apoptosis in MKN1 cells of 11 different heat-killed Bifidobacterium or Lactobacillus strains were examined using the MTT assay or flow cytometry, respectively. Then, BALB/c nude mice xenograft animal models were implanted with human gastric cancer MKN1 cells and orally administered a selected single or a mixture of heat-killed bacterial strains to investigate their inhibitory effect on tumor growth. In addition, the expression of p-Akt, p53, Bax, Bak, cleaved caspase-9, -3, and PARP in the tumor tissues was analyzed using Western blotting assay or immunohistochemistry staining. The results show that heat-killed B. bifidum MG731 (MG731), L. reuteri MG5346 (MG5346), and L. rhamnosus MG5200 (MG5200) induced relatively greater apoptosis than other strains in MKN1 cells. Oral administration of a single dose or a mixture of MG731, MG5346, or MG5200 significantly delayed tumor growth, and MG731 had the most effective anti-tumor effect in the xenograft model. Protein expression of p-Akt, p53, Bax, cleaved caspase-3 and -9, and PARP in tumors derived from the xenograft model correlated with the results of the immunohistochemistry staining.     

Characterization of a Potent,Selective, and Safe Inhibitor,Ac15(Az8)2, in Reversing Multidrug Resistance Mediated by Breast Cancer Resistance Protein (BCRP/ABCG2)

Overexpression of breast cancer resistance transporter (BCRP/ABCG2) in cancers has been explained for the failure of chemotherapy in clinic. Inhibition of the transport activity of BCRP during chemotherapy should reverse multidrug resistance. In this study, a triazole-bridged flavonoid dimer Ac15(Az8)₂ was identified as a potent, nontoxic, and selective BCRP inhibitor. Using BCRP-overexpressing cell lines, its EC₅₀ for reversing BCRP-mediated topotecan resistance was 3 nM in MCF7/MX100 and 72 nM in S1M180 in vitro. Mechanistic studies revealed that Ac15(Az8)₂ restored intracellular drug accumulation by inhibiting BCRP-ATPase activity and drug efflux. It did not down-regulate the cell surface BCRP level to enhance drug retention. It was not a transport substrate of BCRP and showed a non-competitive relationship with DOX in binding to BCRP. A pharmacokinetic study revealed that I.P. administration of 45 mg/kg of Ac15(Az8)₂ resulted in plasma concentration above its EC₅₀ (72 nM) for longer than 24 h. It increased the AUC of topotecan by 2-fold. In an in vivo model of BCRP-overexpressing S1M180 xenograft in Balb/c nude mice, it significantly reversed BCRP-mediated topotecan resistance and inhibited tumor growth by 40% with no serious body weight loss or death incidence. Moreover, it also increased the topotecan level in the S1M180 xenograft by 2-fold. Our results suggest that Ac15(Az8)₂ is a promising candidate for further investigation into combination therapy for treating BCRP-overexpressing cancers.     

RGD特异性结合多肽与重组改构人肿瘤坏死因子融合基因的克隆表达及其抑瘤活性

目的构建RGD多肽与重组改构人肿瘤坏死因子融合基因,进一步提高rmhTNF的临床疗效。方法应用基因重组技术构建了CRGDC与rmhTNF的融合基因,在E.coliDH5α中诱导表达重组蛋白RGDrmhTNF,采用溶菌酶法裂解菌体,裂解上清经硫酸铵沉淀、阴阳离子交换层析纯化后,用S180荷瘤小鼠模型和L929细胞体外杀伤实验测定纯化的重组蛋白RGDrmhTNF的体内、外杀伤活性。结果正确构建了编码RGDrmhTNF融合基因,重组工程菌升温诱导后以部分可溶形式表达RGDrmhTNF,纯化后纯度为96.88%,体外对L929细胞杀伤作用的比活性为3.6×108IUmg,与rmhTNF相当(3.0×108IUmg)。在S180荷瘤小鼠模型中,当RGDrmhTNF剂量为12×105IUkg时,对肿瘤的生长抑制率为84.9%,显著高于相同剂量的rmhTNF(肿瘤抑制率为59.09%,P<0.01)和环磷酰胺组(肿瘤抑制率为71.21%,P<0.01)。结论重组蛋白RGDrmhTNF可以提高rmhTNF的治疗效果。     

Blockade of c-Met-Mediated Signaling Pathways by E7050 Suppresses Growth and Promotes Apoptosis in Multidrug-Resistant Human Uterine Sarcoma Cells

E7050 is a potent inhibitor of c-Met receptor tyrosine kinase and has potential for cancer therapy. However, the underlying molecular mechanism involved in the anti-cancer property of E7050 has not been fully elucidated. The main objective of this study was to investigate the anti-tumor activity of E7050 in multidrug-resistant human uterine sarcoma MES-SA/Dx5 cells in vitro and in vivo, and to define its mechanisms. Our results revealed that E7050 reduced cell viability of MES-SA/Dx5 cells, which was associated with the induction of apoptosis and S phase cell cycle arrest. Additionally, E7050 treatment significantly upregulated the expression of Bax, cleaved PARP, cleaved caspase-3, p21, p53 and cyclin D1, while it downregulated the expression of survivin and cyclin A. On the other hand, the mechanistic study demonstrated that E7050 inhibited the phosphorylation of c-Met, Src, Akt and p38 in HGF-stimulated MES-SA/Dx5 cells. Further in vivo experiments showed that treatment of athymic nude mice carrying MES-SA/Dx5 xenograft tumors with E7050 remarkably suppressed tumor growth. E7050 treatment also decreased the expression of Ki-67 and p-Met, and increased the expression of cleaved caspase-3 in MES-SA/Dx5 tumor sections. Therefore, E7050 is a promising drug that can be developed for the treatment of multidrug-resistant uterine sarcoma.