抗人转铁蛋白受体单克隆抗体体外抗瘤效应研究

目的:探讨抗人转铁蛋白受体(TfR)单克隆抗体(mAb)体外抗瘤效应。方法:采用CFSE染色、PI-AnnexinⅤ染色和PI染色,通过流式细胞术分析抗人TfR mAb对高表达的人肝癌细胞系HepG2和人乳腺癌细胞系MCF-7增殖、凋亡和周期的影响;通过MTT法检测抗人TfR mAb联合化疗药物对HepG2和MCF-7细胞增殖的抑制作用。结果:抗人TfRmAb能抑制HepG2和MCF-7细胞的增殖,并诱导其凋亡和S期阻滞;与化疗药物联用可增强其对肿瘤细胞增殖的抑制作用。结论:抗人TfR mAb具有良好的体外抗瘤效应。

葛根提取物的体外抗瘤作用及流式细胞仪分析

用ＭＴＴ快速测定法及流式细胞仪分析法分析了葛根提取物（ＰＲ）对靶细胞人肝癌ＳＭＭＣ７７２１细胞的抗癌药效，结果葛根提取物有比较强的抗癌活性，与丝裂霉素（ＭＭＣ）联合用药抗癌活性显著增强，流式细胞仪分析细胞分裂周期各时象ＤＮＡ变化显示，ＰＲ作用后Ｓ期细胞明显减少，增殖指数（ＰＩ）降低，并可诱导凋亡峰，提示葛根提取物具有一定抗癌活性。

链球菌制剂SY914体外抗瘤实验研究

为研究ＳＹ９１４β－溶血性链球菌制剂的体外抗瘤作用及作用机制，用ＳＲＢ法对抗癌链球菌制剂ＳＹ９１４作用于人癌细胞株（ＳＧＣ－７９０１，Ａ５４９，ＴＣＡ８１１３，ＫＢ）进行体外抑瘤实验研究，并通过电镜对ＳＹ９１４作用于ＳＧＣ７９１０的过程进行超微结构观察，Ｈ３掺入实验观察ＳＹ９１４对蛋白合成的影响．结果：ＳＹ９１４对４个细胞株具有体外生长抑制作用，ＩＣ５０分别为６２１３（ＳＹ７９０１），１６１７（Ａ５４９），１１５２（ＴＣａ８１１３），１９６（ＫＢ）［ｍｇ／Ｌ］．电镜观察发现ＳＹ９１４作用于瘤细胞后、胞浆内核蛋白体数量明显减少，细胞溶解坏死，Ｈ３掺入实验提示ＳＹ９１４具有显著抑制肿瘤细胞蛋白质合成的作用．结果显示：ＳＹ９１４具有直接抑制肿瘤细胞生长的作用。

PTS(抑瘤仙)体外抗瘤作用研究

目的 研究抗瘤新药PTS(抑瘤仙 )体外对肺癌细胞、耐药肺癌细胞及对正常支气管上皮细胞的作用 ,为其临床应用提供理论依据。方法 以大细胞肺癌株NCI H 460、耐药大细胞肺癌株H 460 /cDDP及正常支气管上皮细胞株 16HBE为实验对象 ,采用 2 4h活细胞跟踪法、MTT法及台盼蓝拒染记录细胞生长曲线法 ,进行PTS的体外抗瘤作用研究 ,同时以DDP(顺铂 )为阳性对照 ,PBS(平衡盐液 )为阴性对照。结果 PTS对受到直接损伤的肺癌细胞及耐药肺癌细胞均有杀伤作用 ,具有抗瘤作用和一定的抗耐药性。与DDP比较 ,PTS有更强的抗瘤作用 ,对于H 460细胞在第 48h 5 0 0mmol的DDP杀伤率为 62 .1% ,1/2 0 0的PTS为 81.3 % ;而PTS对于正常人支气管上皮 16HBE细胞的影响较小 (DDP为 3 9.5 % ,PTS为 2 1.6% ) ,具有较高的选择性。PTS应用后立即起效并达高峰 ,作用持续时间短。结论 PTS体外抗瘤作用明显 。

高免疫原性胶质瘤细胞疫苗的体外抗瘤作用

热休蛋白70（HSP70）与主要组织相容性抗原复合体Ⅰ类分子（MHC—Ⅰ）是参与内源性抗原提呈的两类重要分子。本研究旨在观察模型HSP70及MHC—Ⅰ类分子高表达人多形性胶质母细胞瘤GBM U251细胞疫苗体外诱导的抗瘤作用。

狼毒大戟活性成分体外抑瘤研究

选用体外建株的人肿瘤细胞系U93 7(人恶性组织细胞淋巴瘤细胞株 ) ,Hela(人子宫癌细胞株 ) ,QRH- 770 1(肝癌细胞株 ) ,采用MTT比色法 ,观察了狼毒大戟活性成分W对上述细胞系的影响。结果表明 ,W对

柯里拉京抗肿瘤作用的研究

目的 观察柯里拉京 (Corilagin)体内、外抗肿瘤活性及其量效关系。 方法 体外药效试验用MTT法 ,体内用小鼠移植性肿瘤及裸鼠移植人肿瘤细胞株观察柯里拉京抑瘤活性及与其他化疗药的协同作用。结果 柯里拉京体外对人卵巢癌细胞 (A2 780 )、鼻咽癌细胞 (KB)、骨肉瘤细胞 (OS 732 )和人结肠癌细胞 (HCT 8)IC50 值分别为 0 .2 5 μg/ml、1.2 μg/ml、7.4 2 μg/ml及 9.0 8μg/ml。体内对小鼠移植性肿瘤及裸鼠移植人肿瘤细胞株也显示出较显著抑制作用。结论 柯里拉京体外对A2 780、KB、OS 732和HCT 8人结肠癌细胞有明显的细胞毒活性 ,体内对小鼠肝癌H2 2 ,裸鼠移植人OS 732、HCT 8也显示出较好的抑制作用。

A Bioactive Diterpene from the Chinese Herb Aster souliei

Aster souliei Franch, (Compositae) is a. herbaceous plant distributed innorth China. It has been used in folk medicine as antipyretic, detoxicant, expectorant andantitussive. In an effort to find biologically active components from Chinese medicinal plants ' ,we have examined the aerial parts of this herb, leading to the isolation of a clerodane-typediterpene, 18, 19-dihydroxy-5α, 10β-neo-cleroda-3, 13 (l4)-dien-16, 15-butenolide (1). In thispaper we report the structural elucidation, and the antitumor and antibacterial activities of thiscompound. It was found that 1 possesses moderate cytotoxicity against human leukemia cells (HL-60)and activity against microorganisms.

The Anti—tumor Effects of an Anti—CD71 Chimeric Antibody in Vitro and Its Distribution in a Tumor Xenograft Model

Objective To investigate the anti-tumor effects in vitro and in vivo distribution of the human/murine chimeric antibody (D2C). Methods The CD71 positive target cells (K562, GEM and SMMC7721) and the effector cells, freshly isolated human PBMC, with the ratio of target cells to effector cells 1:50, were incubated in various dilutions of D2C antibody ( Ab) . Antibody dependent cytotoxicity (AD-CC) was tested by using an LDH-release assay. Instead of effector cells, complement was added to the target cells (GEM, SMMC-7721) with various dilutions of D2C Ab. A method of counting death cells was used in complement dependent cytotoxicity (CDC) assay. Tumor localization and distribution of the chimeric antibody (D2C) were observed by labeling the chimeric Ab with radioiodine(131I) and injecting it into nude mice (Balb/c nu/nu) transplanted with human hepatocellular carcinoma cells (SMMC-7721).Results A significant ADCC was observed with the increased concentration of the D2C Ab. Cytolysis of CD71-positive target cells by the D2C Ab was found in the presence of fresh rabbit complement. Labeled D2C administered by intraperitoneal as well as tumor regional injection, was visualized by SPECT. The distribution of D2C Ab in murine organs and tissues showed that non-specific binding was lower following tumor regional administration than when the antibody was administered by an intraperitoneal injection. The human/murine chimeric antibody (D2C) has in vitro anti-tumor effects and can exert its effects in specific tumor localization. Its distribution and local effects in vivo can be detected by radioimmunoimaging.Conclusion CD71 human/murine chimeric antibody showed marked killing of tumor cells in vitro, and specific recognition and high affinity binding to tumor tissue in vivo

Jinshuixian in combination with radiotherapy suppresses the proliferation of A549 cells by inhibiting the expression of VEGF

Objective： The aim of our study was to investigate whether Jinshuixian has the abilities of inhibiting tumor growth and radio-sensitivity effect. Methods： Cultured lung A549 ceils were randomly divided into 6 groups： normal control group （NC）, the Jinshuixian group （JSX）, radiotherapy （RT）, JSX for the first day and the next day followed by RT group （JSX~ RT）, RT for the first day and the next day followed by JSX group （RT→JSX） and RT ＋ JSX concomitantly group （JSX ＋ RT）. MTT was applied to measure the cell viability, RT-PCR and ELISA were used to test the expression of mRNA and protein of VEGF. Results： The proliferation of A549 cells were inhibited in JSX and combined groups and the inhibiting effects were time dependent. The expression of VEGF in RT group was increased, however, VEGF in JSX and combination groups were largely decreased over time when compared to NC group. Results in JSX→RT, RT→JSX and JSX ＋ RT groups did not achieve significantly differences. Conclusion： JSX has the ability of anti-tumor growth in vitro accompanied down-regulating of VEGF, especially when combined with radiotherapy, and its effect is time-dependent. However, more studies in vivo and in vitro are needed to further supporting these effects.

In vitro antitumor activity and targeted sites of two novel platinum-based(II) complexes on SW620 colon cancer cell line

Objective:The aim of our study was to evaluate the in vitro antitumor activity of two novel platinum-based(II) complexes(2.3-pyridinedicarboxylic acid dehydrate platinum and 2.3-pyrazinedicarboxylic acid dehydrate platinum),which were concurrently provided with hydrophilic carboxyl group and lipophilic pyrazinyl or pyridyl group,on SW620 colorectal cancer cell line and the impact of the two compounds on the cell cycle and apoptosis of the cells when compared with the oxaliplatin,desiring the new ligand combined with hydrophilic and lipophilic properties would facilitate the transportation and transmembrane of the drugs,showing a better antitumor activity.Methods:After SW620 cells were treated with different doses of the three platinum-based agents for 24,48 and 72 h,the cell proliferation inhibition rate was determined using methyl thiazolyl tetrazolium(MTT) assay;the morphology of cells were evaluated under inverted microscope;the changes in cell cycle were determined using flow cytometry;the percent apoptosis was measured using Annexin V/PI double staining and the micromorphology of the cells after drug exposure was evaluated using scanning electron microscopy.Results:The evaluation on the proliferation inhibition rate revealed that the three platinum-based agents inhibited the SW620 cells in a time-and dose-dependent manner and showed different strengths as pyridine > pyrazine > Oxa.Under optical microscope,the morphological changes such as cell shrinkage,round cells and dead cells were frequently observed after drug exposure.Cell cycle determination showed that all of the three agents could function to block the cells converting from phase S to phase G2M.Apoptosis evaluation revealed that the three agents promoted the apoptosis of SW620 cells in a time-and dose-dependent manner and showed different strengths as pyridine > pyrazine > Oxa.Typical early and late apoptotic morphological changes could be detected during electron microscopy.Conclusion:The two novel platinum-based(II) complexes showed a stronger antitumor effect on SW620 cells than oxaliplatin,with the targeted site at a certain phase of cell cycle and apoptosis.

Synthesis of Novel Series of Benzothieno [2,3-d] Pyrimidine Derivatives, Promising Anticancer Agents

An extension of the authors＇ previous discovery of in vitro antitumor activity of substituted thino [2,3-d] prymidine derivatives is reported. The synthesis of some new spirothino [2,3-d] prymidine （4a-f）, imidazolidin, substituted prymidinyl and substituted thiazolidine thino [2,3-d] prymidine derivatives have been described. Thirteen of the obtained compounds were selected by the NCI and evaluated for their in vitro anticancer activity. Seven of the investigated compounds, 4a, 8a, 9a, （12a, b）, 14a and 15a, displayed high anticancer activity in the primary assay. These compounds have been selected for a full anticancer screening against a 60-cell panel assay where they showed non-selective broad spectrum and promising activity against all cancer cell lines. Compounds 12a and 12b proved to be the active members in this study compared to 5-fluorouracil and cyclophosphamide as reference drugs, respectively. Compounds 12a and 12b were identified as promising lead compounds, evaluated for their in-vitro antitumor activity.

Anticancer effect and enhanced chemotherapy potential of resveratrol in human pancreatic cancer cell lines

Objective Gemcitabine, the only approved drug for the treatment of pancreatic cancer, is not very effective. Novel and effective cancer chemopreventive agents are urgently needed. Recently, emerging studies determined resveratrol possessed anticancer effects on various cancer cells. We explored the anticancer effect of resveratrol in pancreatic cancer cells and investigated the involved moleculars of action. We also examined whether resveratrol enhanced antitumor activity of gemcitabine in vitro.Methods Proliferation inhibition was assessed by cell count kit-8 assay. Cell cycle phase distribution and apoptotic cells were measured by flow cytometric analysis. We determined the expression of bcl-2, cyclinD1, and activation of caspases-3 and poly(ADP-ribose) polymerase1 proteins used Western blot analysis.Results Resveratrol inhibited the proliferation of three pancreatic cancer cell lines in a dose dependent fashion, and induced accumulation of cells at the G1 phase as well as apoptosis. Our data also demonstrated that resveratrol enhanced gemcitabine-induced apoptosis in pancreatic cancer cells. In addition, resveratrol inhibited the expression of cyclinD1, bcl-2, and induced activation of caspase-3 and poly(ADPribose) polymerase1. Conclusion Our results suggested that resveratrol might be not only a potential regimen, but also an effective chemosensitizer for the chemotherapy of pancreatic cancer.

Isolation and Characterization of Exosomes Derived from Tumor Cells Genetically Expressing Model Antigen

Tumor cell-derived exosomes have been proposed as non-cellular nanomeric vaccine which could induce potent anti- tumor immune response in mice. In order to develop the protocols to prepare tumor cell-derived exosomes for basic research and clinical trail, we isolated exosomes from ovalbumin (OVA)-expressing thymoma cells EG.7-OVA by various preparation methods. We demonstrate the non-sedimentation method is simple, rapid, efficient with higher yield and purity of exosomes. EG.7-OVA-derived exosomes are 40-100 nm in diameter sequestered by lipid bi-layer, and contain rich heat shock protein (HSP) and OVA. The result of the size distribution determination is consistent with the calculation by the visual microscopic inspection, with 90.4% particles at the range of 50-90 nm. Moreover, as a model antigen of the EG.7 cells, OVA concentra- tion in EG.7-derived exosomes can be regarded as a good quality control parameter. Therefore, we have established a platform to efficiently prepare exosomes for tumor immunotherapy.

The cellular uptake and anti-tumor activity of conjugated linoleic acid-paclitaxelloaded iRGD-modified lysolipid-containing thermosensitive liposomes

In the present study, we prepared the iRGD-modified lysolipid-containing thermosensitive liposomes(LTSL) containing conjugated linoleic acid-paclitaxel(CLA-PTX), also known as iRGD-LTSL-CLA-PTX. The in vitro cellular uptake and in vitro cytotoxicity of iRGD-LTSL-CLA-PTX were evaluated in B16-F10 melanoma cells. The in vivo anti-tumor effect of i RGD-LTSL-CLA-PTX was investigated using B16-F10 tumor-bearing C57BL/6 mice. The results of the cellular uptake experiment indicated that the increased cellular uptake of CLA-PTX in the iRGD-LTSL-CLA-PTX-treated groups was 2.05-, 3.31-or 4.83-fold compared with that in the SSL-CLA-PTX group after a 2-, 4-or 6-h incubation at 42 °C, respectively. The in vivo antitumor results showed that iRGD-LTSL-CLA-PTX/heat significantly inhibited the growth of B16-F10 tumors compared with the CLA-PTX solution(LTSL-CLA-PTX, LTSL-CLA-PTX/heat and iRGD-LTSL-CLA-PTX)(P<0.01). In conclusion, the antitumor effect of iRGD-LTSL-CLA-PTX was confirmed on B16-F10 melanoma in vitro and in vivo, which was induced by both the effect of iRGD and LTSL.

Design, synthesis and in vitro antitumor evaluation of novel diaryl urea derivatives bearing sulfonamide moiety

A novel series of diaryl urea derivatives bearing sulfonamide moiety have been designed and synthesized. Their in vitro antitumor effect against human cancer cell lines MX-1, A375, HepG2, Ketr3 and HT-29 was screened and evaluated by the standard MTT assay with sorafenib as the positive control. Some of the compounds showed significant inhibitory activity against multiple cell lines compared to sorafenib. In particular, 2,6-dimethyl-4-{6-[3-（4-chloro-3-（trifluoromethyl）phenyl）urealnaphthalen- 2-yllsulfonyl morpholine （10d） was found to be the most potent against A375, HepG2 and Ketr3 with ICs0 values of 0.65-0.97 μmol/L, which were 5-20-fold more potent than sorafenib. Compound 10d emerged as a valuable lead for further optimization.

The in vitro and in vivo anti-tumor effects of MTX-Fe_3O_4-PLLA-PEG-PLLA microspheres prepared by suspension-enhanced dispersion by supercritical CO_2

The in vitro and in vivo anti-tumor efficacy of methotrexate-loaded Fe3O4-poly-L-lactide-poly(ethylene glycol)-poly-L-lactide magnetic composite microspheres(MTX-Fe3O4-PLLA-PEG-PLLA MCMs,MMCMs),which were produced by co-precipitation(C)and microencapsulation(M)in a supercritical process,was evaluated at various levels:cellular,molecular,and integrated.The results at the cellular level indicate that MMCMs(M)show a better anti-proliferation activity than raw MTX and could induce morphological changes of cells undergoing apoptosis.At the molecular level,MMCMs(M)lead to a significantly higher relative mRNA expression of bax/bcl-2 and caspase-3 than MMCMs(C)at 10μg mL-1(P<0.01);and the pro-caspase-3protein expression measured by Western blot analysis also demonstrates that MMCMs(M)can effectively activate pro-caspase-3.At the integrated level,mice bearing a sarcoma-180 tumor are used;in vivo anti-tumor activity tests reveal that MMCMs(M)with magnetic induction display a much higher tumor suppression rate and lower toxicity than raw MTX.Pharmacokinetic studies show that MMCMs(M)with magnetic induction significantly increase the accumulation of MTX in the tumor tissue compared with the other treatments.These results suggest that the MMCMs(M)prepared by the SpEDS process have great potential to play a positive role in the magnetic targeted therapy field.