中药香加皮与其易混品体外抑瘤效果研究

背景与目的:比较中药香加皮及其易混品水提取物的体外抗肿瘤效果。材料与方法:采用四甲基偶氮唑蓝(3_[4,5_dimethylthiazol_2_yl]_2,5diphenyltetrazoliumbromide,MTT)法测定了中药香加皮及其易混品水提取物对多种不同组织来源的肿瘤细胞增殖反应的影响。结果:香加皮水提取物对人红白血病细胞株K562,人胃癌细胞株BGC_823,人食管癌细胞株TE_13的增殖有明显的抑制作用,且量效关系明显;对实验组其它肿瘤细胞株也有较高抑制率。香加皮易混品水提取物对肿瘤细胞虽也有一定的抑制作用,但其强度明显弱于香加皮水提取物。结论:香加皮水提取物对不同组织来源的肿瘤细胞株有广泛的抑制作用,其抗肿瘤作用明显高于其易混品,香加皮作为抗肿瘤药物有待进一步开发。     

榄香烯乳注射液抗癌作用的研究

目的探讨榄香烯乳注射液对小鼠移植性肿瘤及肿瘤细胞株的抗癌作用.方法体内抑瘤试验:选肝癌H22、肉瘤S-180、艾氏腹水癌等瘤株.腹腔注射给药,观察记录对腹水癌的生命延长作用及对实体瘤的抑制作用.体外细胞毒试验:选用K562白血病、Hela宫颈癌细胞株,采用MTT法,计算对肿瘤细胞株平均细胞生长抑制率.结果榄香烯乳注射液能明显延长腹水型荷瘤小鼠的生存时间.对K562白血病、Hela宫颈癌细胞株有一定的抑制作用.结论榄香烯乳注射液对肿瘤细胞体内抑瘤试验、体外细胞毒试验均有明显的抑制作用.     

双自杀基因CD和TK对K562细胞体内外杀伤作用的实验研究

目的:探讨双自杀基因CD和TK对K562细胞的体内外抑制作用及前体药物对肿瘤的杀伤作用。方法:将目的基因转染入K562细胞,MTT法观察细胞在体内外的增殖状况及5-FC/GCV对转染细胞的杀伤作用,电子显微镜观察其超微结构变化。将K562/CDgly TK和K562细胞接种于裸鼠皮下,观察各种肿瘤细胞在体内的成瘤情况及对前体药物治疗的敏感性。结果:单独使用GCV或5-FC对K562及K562/CDgly TK细胞产生明显的杀伤作用,联合应用该2种药物对肿瘤细胞的杀伤作用更强。将K562细胞和K562/CDgly TK细胞接种于小鼠皮下后小鼠成瘤率为100%,GCV或5-FC可明显抑制裸鼠体内的肿瘤形成,联合应用GCV和5-FC治疗K562/CDglyTK细胞在小鼠体内形成的肿瘤,较单独应用GCV和5-FC及对照组小鼠形成的肿瘤体积明显缩小,生存期也明显延长。结论:双自杀基因在体内外对K562细胞均有明显的抑制作用,可增加前体药物GCV和5-FC对瘤细胞的杀伤率。     

4-1BBL基因修饰colon26细胞的体内抗肿瘤作用

背景与目的:研究4-1BBL基因修饰的小鼠结肠癌细胞瘤苗在小鼠体内诱导产生免疫反应及抗肿瘤效应的能力. 材料与方法:采用脂质体介导法将真核表达质粒pMKIT/4-1BBL导人小鼠结肠癌细胞colon26,经G418筛选后获得4-1BBL稳定高表达细胞克隆colon26/4-1BBL和空载体细胞克隆colon26/pMKITneo,以丝裂霉素C处理,制成癌细胞瘤苗.将野生型colon26、eolon26/pMKITneo和colon26/4-IBBL细胞分别接种于BALB/c小鼠右侧背部皮下,观察细胞致瘤性.取MMC处理的colon26细胞、colon26/pMKITneo细胞和colon26/4-1BBL细胞,分别腹腔免疫BALB/c小鼠,采用改良MTT法测定CTL和NK细胞杀伤活性,取血清用ELISA法测定IL-2、IFN-γ含量.于BALB/c小鼠皮下接种colon26细胞制备早期癌模型,第2 d腹腔注射colon26/4-1BBL瘤苗,7 d后再注射1次,观察肿瘤生长情况.结果:与野生型colon26和colon26/pMKITneo细胞相比,colon26/4-1BBL细胞在小鼠体内的致瘤性明显下降,表现为肿瘤结节出现时间延迟,肿瘤体积明显小于对照组(P＜0.05).MMC处理的肿瘤细胞瘤苗免疫小鼠后,colon26/4-1BBL组小鼠CTL和NK细胞的杀伤活性明显增高(P＜0.05);血清IL-2和IFN-γ的含量明显增高(P＜0.01);而且能对亲本肿瘤细胞colon26产生免疫保护作用,大部分小鼠能保持无瘤状态长期存活(100 d以上).colon26/4-1BBL瘤苗治疗后,部分小鼠保持无瘤生存,成瘤小鼠肿瘤生长缓慢,小鼠生存期明显延长(＞70 d).结论:4-1BBL基因修饰的小鼠结肠癌细胞瘤苗能显著增强小鼠的免疫功能,并能产生显著的抗肿瘤作用.     

去氢骆驼蓬硷抗癌作用的研究

目的研究去氢骆驼蓬硷对小鼠移植性肿瘤及肿瘤细胞株的抑制作用.方法体内抑瘤试验选用肉瘤S-180、肝癌H22、艾氏腹水癌等瘤株腹腔注射给药,观察对实体瘤的抑制作用及对腹水癌的生命延长作用.体外细胞毒试验选用K562、Hela细胞株,采用MTT法,计算对肿瘤细胞株平均细胞生长抑制率.结果去氢骆驼蓬硷对肉瘤S-180及肝癌H22的抑制率具有非常显著性差异(P＜0.01)对艾氏腹水癌作用不明显.对K 562细胞生长半数抑制浓度IC50为3.8μg/ml,对Hela细胞生长半数抑制浓度IC50为13.1μg/ml.结论去氢骆驼蓬硷对肿瘤细胞体内抑瘤试验、体外细胞毒试验均有明显的抑制作用.     

消化腺提取物对肿瘤细胞的抑制作用

背景与目的:研究消化腺提取物对小鼠移植瘤模型的抑制作用.材料与方法:将肝癌H22、肉瘤S180和人胃癌MGC-803细胞株移植到小鼠或裸鼠体内,建立荷瘤动物模型,连续灌胃给予消化腺提取物(EDG)后,分期测量肿瘤体积和瘤重.结果:给予EDG后,荷瘤鼠的瘤体生长速度明显减慢,相对肿瘤体积明显缩小(P＜0.01);肿瘤增殖率均未超出60%;肿瘤体积抑制率达到43.7%以上;EDG各剂量组的瘤体重量明显减轻.结论:消化腺提取物对荷瘤小鼠具有明显的抑制肿瘤生长作用.     

香加皮水提物抑制诱导红白血病细胞K562分化的初步研究

背景与目的： 研究中药香加皮水提取物(cortex periplocae extract,CPE) 诱导体外培养人红白血病细胞株K562分化的作用。 材料与方法： 观察人红白血病细胞株K562与CPE体外共培养时的形态学变化;同时采用比色法检测不同浓度CPE诱导人红白血病细胞株K562产生Hb含量的变化。 结果： 与阴性对照相比,CPE可以诱导K562细胞发生坏死的形态学变化。与CPE共培养后K562细胞株Hb的产生明显减少。 结论： CPE 对K562肿瘤细胞只具有杀伤作用而不能诱导K562肿瘤细胞的分化。     

胎盘多肽对肿瘤细胞生长抑制作用研究

目的　探讨胎盘多肽 (PPs)对小鼠肿瘤细胞体内、外生长的影响。方法　染料排斥试验及肿瘤细胞体内种植实验。结果　PPs在体外对肿瘤细胞无明显的直接杀伤作用 (P >0 0 5 ) ,但体内肿瘤种植实验中给药组H2 2 腹水瘤小鼠生命延长率为 70 % ,S180 实体瘤生长抑制率为 6 5 % ,与对照组相比实验组荷瘤小鼠生存时间有显著差异 (P <0 0 1 )且肿瘤的形成也有明显的抑制作用 (P <0 0 1 )。结论　PPs在体内有明显的抑瘤作用     

冻融抗原冲击致敏的树突状细胞对结肠癌小鼠的治疗作用

目的 :研究冻融的肿瘤抗原冲击致敏的骨髓来源的树突状细胞 (DC)对结肠癌小鼠是否具有治疗作用。方法 :用小鼠结肠癌细胞株C2 6冻融抗原体外冲击致敏BALB/c小鼠骨髓来源的DC ,观察其体外刺激荷瘤小鼠脾细胞增殖的能力及其诱导的CTL对C2 6肿瘤细胞的杀伤活性 ;体内以 3× 10 5DC /只一次或多次接种于已荷瘤 10d结肠癌的小鼠同侧腹股沟皮下 ,观察抗原冲击的DC对肿瘤生长的抑制作用以及对荷瘤小鼠生存期的影响。结果 :体外抗原冲击致敏的DC能显著刺激荷瘤小鼠脾脏T细胞增殖 ,其诱导的CTL对C2 6肿瘤细胞具有显著的杀伤作用 ,在效靶比为 10∶1,5∶1,2 .5∶1时其杀伤率分别 88 1% ,71.4%和 5 0 .0 % ;抗原冲击致敏的DC体内多次皮下免疫后对肿瘤的生长具有显著的抑制作用 ,能显著延长荷瘤小鼠的生存期。一次性DC体内免疫接种对荷瘤小鼠的治疗作用不明显。结论 :肿瘤细胞冻融抗原体外冲击致敏的DC多次皮下免疫对结肠癌小鼠具有显著的治疗效果     

千金子甲醇提取物抗肿瘤作用的实验研究

 目的　观察千金子甲醇提取物体内、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 ,观察千金子甲醇提取物的抑瘤活性。结果　千金子甲醇提取物体外对人宫颈癌细胞 (Hela)、人红白血病细胞 (K562 )、人单核细胞性白血病细胞 (U93 7)、人急性淋巴细胞性白血病细胞 (HL60 )和人肝癌细胞 (HepG2 )的IC50 值分别为 15 .5 μg/ml、13.1μg/ml、10 .5 μg/ml、17.5 μg/ml、2 9 .6 μg/ml;体内对小鼠移植性肿瘤细胞株也显示出较显著抑制作用。 结论　千金子甲醇提取物体外对Hela、K562 、U93 7、HL60 、HepG2 有明显的细胞毒活性 ,体内对小鼠肉瘤 180 (S180 )和艾氏腹水癌 (EAC)也显示出较好的抑制作用     

CIK的体外增殖及体内外杀瘤活性的实验研究

目的:从人骨髓造血前体细胞体外培养扩增树突状细胞(dendritic cells,DCs),测定其表型及T细胞刺激活性.方法:采用Mini-MACS分离技术,从正常人骨髓、脐血分离CD34~ 造血干细胞,体外以重组hGM-CSF,hTNF-α,hIL-3诱导培养2周,流式细胞术检测扩增细胞的表面表型及细胞内IL-12的表达,体外同种混合淋巴细胞反应检测扩增DCs的T细胞刺激活性.结果:从正常人骨髓、脐血分离得到高纯度(>90%)的CD34~ 造血干细胞,经重组hGM-CSF,hTNF-α的共同诱导培养,扩增得到大量DCs,加人hIL-3可以进一步增加DCs产量;FACS检测表明,扩增的DCs表达HLA-DR,CD40,CD54,CD80,CD86分子,细胞内有hIL-12的P35,P40亚基的表达;与外周血单核细胞培养生成的DCs相比,由CD34~ 干细胞扩增的DCs具有更强的激发同种T细胞增殖的能力.结论:人CD34~ 干细胞体外经诱导培养,可以生成大量功能成熟的DCs,从而为进一步开展DCs的基础及临床研究打下了基础.     

人肺癌细胞NHE—1基因片段的克隆及其反义表达载体的构建

目的:动态观察CIK(cytokine induced killer)细胞的体外增殖,体外的细胞毒活性,及通过动物实验研究其体内的抗肿瘤作用.方法:通过提取健康供血者的PBMC,第0天加入γ-IFN,第1天加入IL-2、抗-CD3单抗和IL-1培养CIK细胞;在流式细胞仪上做动态培养物的表型分析;与LAK细胞作对比,分别用MIT法测定其体外细胞毒活性及对S180荷瘤鼠的体内抗肿瘤作用.结果:CIK细胞在培养2周后获得大量增殖,表型分析表明,CIK细胞属异质性细胞群,在培养的过程中,群体的CD3~ CD56~ 细胞大量扩增达1000多倍,是CIK细胞的主要效应细胞;实验证明,CIK细胞的体外细胞毒活性及对S180荷瘤鼠的体内抗肿瘤作用均强于LAK细胞;其较强的体内抗癌活性可能与荷瘤鼠主体内T细胞活化有关.结论:CIK细胞是一种强于LAK细胞的、新型、高效、具有广谱杀瘤活力的免疫活性细胞.     

Anti-tumor Effects of Penfluridol through Dysregulation of Cholesterol Homeostasis

Background: Psychiatric patients appear to be at lower risk of cancer. Some antipsychotic drugs might haveinhibitory effects on tumor growth, including penfluridol, a strong agent. To test this, we conducted a studyto determine whether penfluridol exerts cytotoxic effects on tumor cells and, if so, to explore its anti-tumormechanisms. Methods: Growth inhibition of mouse cancer cell lines by penfluridol was determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxic activity was determined byclonogenic cell survival and trypan blue assays. Animal tumor models of these cancer cells were established andto evaluate penfluridol for its anti-tumor efficacy in vivo. Unesterified cholesterol in cancer cells was examined byfilipin staining. Serum total cholesterol and tumor total cholesterol were detected using the cholesterol oxidase/paminophenazone(CHOD-PAP) method. Results: Penfluridol inhibited the proliferation of B16 melanoma (B16/F10), LL/2 lung carcinoma (LL/2), CT26 colon carcinoma (CT26) and 4T1 breast cancer (4T1) cells in vitro. Invivo penfluridol was particularly effective at inhibiting LL/2 lung tumor growth, and obviously prolonged thesurvival time of mice bearing LL/2 lung tumors implanted subcutaneously. Accumulated unesterified cholesterolwas found in all of the cancer cells treated with penfluridol, and this effect was most evident in LL/2, 4T1 andCT26 cells. No significant difference in serum cholesterol levels was found between the normal saline-treatedmice and the penfluridol-treated mice. However, a dose-dependent decrease of total cholesterol in tumor tissueswas observed in penfluridol-treated mice, which was most evident in B16/F10-, LL/2-, and 4T1-tumor-bearingmice. Conclusion: Our results suggested that penfluridol is not only cytotoxic to cancer cells in vitro but canalso inhibit tumor growth in vivo. Dysregulation of cholesterol homeostasis by penfluridol may be involved inits anti-tumor mechanisms.     

γ-synuclein对结肠癌细胞SW1116体外及体内增殖潜能的影响

目的:研究γ-synuclein对人结肠癌细胞系体外及体内增殖潜能的影响.方法:利用真核表达载体pIRES2-EGFP和RNA干扰载体pGCsi-U6/neo/GFP构建针对γ-synuclein的重组质粒,并转染至人结肠癌细胞株SW1116,经G418筛选出稳定转染细胞株.应用CCK8法检测细胞增殖活性;软琼脂克隆法检测细胞克隆形成能力;裸鼠皮下成瘤实验检测细胞在裸鼠体内的成瘤能力.结果:经测序以及RT-PCR和Westernblot验证,重组质粒成功构建,并能够过表达或抑制γ-synuclein基因和蛋白.结果显示,y-synuclein受抑制后,SW1116细胞的增殖数目从48h至120h持续显著减少(P＜0.05),克隆形成率亦显著降低(P＜0.05).SW1116细胞的增殖和克隆形成能力与其γ-synuclein的表达水平呈正相关.过表达γ-synuclein能够部分增强SW1116细胞的增殖和克隆形成能力,但差异无统计学意义(P＞0.05).体内实验也有类似结果,过表达γ-synuclein能够稍微加快SW1116细胞瘤体生长速度,但直至第27天,其生长速度才显著加快(P＜0.05).γ-synuclein受抑制后,SW11 16细胞裸鼠皮下成瘤速度从第6天至第30天显著减慢(P＜0.05).结论:γ-synuclein与结肠癌细胞体外及体内的增殖潜能密切相关.     

PE, a new sulfated saponin from sea cucumber, exhibits anti-angiogenic and anti-tumor activities in vitro and in vivo

Here, we examined the in vitro and in vivo anti-angiogenesis and anti-tumor activities of PE, a new marine-derived compound. Inhibition of angiogenesis was assessed in vitro using proliferation, migration, adhesion, tube-formation and apoptosis assays in PE-treated HMECs and HUVECs. In vivo, CAM assays were used to assess inhibition effect of PE on physiological angiogenesis, and immunofluorescent microscopy was used to examine tumor microvessel density and apoptosis in PE-treated mouse tumor models. Finally, Western blotting analyses were performed to examine the effect of PE on VEGF signaling in HMECs. The results showed that PE inhibited proliferation of HMECs and HUVECs with IC50 values of 2.22 +/- 0.31 microM and 1.98 +/- 0.32 microM, induced endothelial cell apoptosis at concentrations <2 microM, induced dose-dependent suppression of cell migration, cell adhesion and tube formation in HMECs and HUVECs, and showed anti-proliferative activities against several tumor cell lines (IC50 values of approximately 4 microM). In vivo, PE (5 nM/egg) suppressed spontaneous angiogenesis in our CAM assay, and induced marked growth inhibition in mouse sarcoma 180 and hepatoma 22 models. Specifically, PE treatment reduced mouse sarcoma 180 tumor volume by triggering apoptosis of both tumor and tumor-associated endothelial cells, preferentially targeting on endothelial cells comparable with tumor cells. Finally, PE treatment suppressed the active (phosphorylated) forms of VEGFR2, Akt, ERK, FAK and paxillin, which are involved in endothelial cell survival, proliferation, adhesion and migration. Our results indicate that PE exerts an anti-angiogenic activity associated with inhibition of VEGFR2 signaling, and an anti-tumor activity associated with decreased proliferation of tumor cells and increased apoptosis of both endothelial cells and tumor cells.     

木蹄层孔菌乙醇提取物对肿瘤细胞的抑制作用

背景与目的 :研究木蹄层孔菌乙醇提取物(ethanolextractofFomesfomentarius,EEF)体内外对肿瘤细胞的抑制作用。 材料与方法 :噻唑蓝(3_[4,5_dimethylthiazol_2_yl]_2,5diphenyltetrazoliumbroide,MTT)法测定EEF对体外培养的HeLa细胞的生长的影响 ;流式细胞仪检测其体外对S180 细胞凋亡率的影响 ;建立腹水型S180肉瘤模型 ,测定EEF对荷瘤鼠生命延长率的影响 ;通过试剂盒检测EEF对正常小鼠几个重要器官丙二醛(Malondialdehyde,MDA)含量的影响。 结果 :EEF对体外生长的HeLa细胞有明显的抑制作用 ;能显著提高S180 细胞的凋亡率及荷瘤鼠的生存天数 ;能明显降低正常鼠肝脏、肾脏MDA含量。 结论 :EEF对肿瘤细胞有很强的抑制作用。     

THE BIOLOGICAL CHARACTERISTICS OF ADENOVIRUS-MEDIATED IL-18 GENE-MODIFIED MURINE COLORECTAL ADENOCARCINOMA CELL IN VIVO AND IN VITRO

Interleukin 18 (IL-18) has multiple biological activities, such as promoting the generation of Th1 cytokines and GM-CSF, activating NK cells and CTL, which contributes to its anti-tumor activity.[1(5] So IL-18 might have promising application in the immunotherapy and gene therapy of cancer. To confirm the anti-tumor activity of IL-18, we constructed the recombinant adenovirus encoding IL-18 gene, observed preliminarily the biological characteristics of IL-18-modified murine colorectal …     

Successful adoptive immunotherapy with OK432-inducible activated natural killer cells on tumor-bearing mice

We had demonstrated that the NK cell mediated cytotoxicity of murine spleen cells could be augmented by in vivo prime and subsequent in vitro challenge with the streptococcal preparation OK432, and the cell surface phenotype of induced killer cells was Thy 1+, asialo GM1+, suggesting the activated NK cells (OK-NK cell). The culture supernatants of spleen cells with OK432 possessed the activity of IL-2 and IFN-gamma, and the IL-2 played a major role to induce the OK-NK cells via the production of IFN-gamma. In this study, we examined the effect of adoptive transfer of OK-NK cells on tumor-bearing mice. The mice were implanted SP2 myeloma cells intraperitoneally (i.p.), or C26 colon adenocarcinoma cells subcutaneously (s.c.) to make the models of peritonitis carcinomatosa or solid tumor, and the OK-NK cells were transferred i.p. or i.t., adoptively. By the adoptive transfer of OK-NK cells, the 92% of mice bearing SP2-tumor had be cured. The tumor growth of C26-solid tumor was inhibited, and the survival rate of mice bearing C26-tumor was increased, significantly. The intratumoral remnants of 125I-labelled OK-NK cells were 61.27 and 8% after intratumoral transfer, respectively. By multiple transfer of OK-NK cells the anti-tumor effect was more augmented than that of a single transfer. Thus we recognized the anti-tumor effect of adoptive transfer of OK-NK cells on tumor-bearing mice, and suggested that OK-NK cells could be useful for the therapy of cancer patients.     

4-1BBL修饰小鼠结肠癌疫苗体外诱导抗肿瘤免疫反应的研究

背景与目的:研究4-1BBL(4-1BB Ligand,即CD137配体)转基因小鼠结肠癌细胞瘤苗体外诱导细胞毒性T淋巴细胞(Cytotoxic T lymphocytes,CTL)特异性杀伤活性及刺激淋巴细胞产生细胞因子的作用.材料与方法:采用脂质体介导法将真核表达质粒pMKITneo/4-1BBL导入小鼠结肠癌colon26细胞,经G418筛选后获得4-1BBL高表达克隆,用丝裂霉素C(MMC)处理后,制成肿瘤细胞瘤苗,与经体外诱导的同系小鼠CTL共同培养,测定对CTL特异性杀伤活性及对脾细胞产生细胞因子(IL-2、IL.-4和IFN-γ)的影响.结果:转染4-1BBL的colon26细胞高表达4-1BBL蛋白,将该细胞经MMC处理后制成的瘤苗,与野生型colon26细胞相比,对CTL特异性杀伤亲本肿瘤细胞的作用明显增强(P＜0.01),但CTL对非亲本肿瘤细胞的杀伤作用无明显影响(P＞0.05);该瘤苗在体外能显著增强脾细胞分泌细胞因子(IL-2、IL-4和IFN-γ)的能力(P＜0.01).结论:4-1BBL转基因小鼠结肠癌细胞瘤苗能有效诱导抗结肠癌免疫反应.