Influence of granulocyte-macrophage colonystimulating factor and tumor necrosis factor on anti-hepatoma activities of human dendritic cells

INTRODUCTIONDendritic cells (DCs) play a key regulatory role inantitumor immunity,especially in its immuneaccessory role via MHC-Ⅰ molecules.We haverecently reported that DCs were able to enhance thekilling activity of Lymphokine and PHA activatedkiller (LPAK) cells in vitro.In the presentstudy,we evaluated the effects of GM-CSF andTNF upon antitumor activities of freshly

Anti-hepatoma Effect of DC2.4 Cells Transfected with Tumor-Associated Antigen Cdc25C In Vitro

Objective Cell division cyclin 25 homolog C(Cdc25C)is a tumor-associated antigen candidate gene,and this may be used as an effective target in cancer treatment.The present study aims to evaluate the lysis effect of cytotoxic T lymphocytes(CTLs)induced by dendritic cell line DC2.4 overexpressing Cdc25C,and the feasibility of Cdc25C as a component in hepatoma immunotherapy.Methods The mouse Cdc25C gene was ligated into a lentiviral vector,and transfected into DC2.4 cells.The DC2.4 cell phenotype and cytokine secretion were determined by flow cytometry and ELISA,respectively.CD8^(+)T cells were sorted from the spleens of C57BL/6 mice using a magnetic bead sorting kit obtained from Miltenyi Biotech,Germany,and co-cultured with DC2.4 cells for one week as effector cells.Then,IL-2,granzyme B and perforin were detected in the CTL culture medium by ELISA.Next,time-resolved fluorescence immunoassay was used to detect the immune killing effect of Cdc25C-specific CTLs on target cells.Meanwhile,the effect of blocking MHC-I sites on target cells with a monoclonal anti-MHC-I antibody was evaluated.Results The results revealed that Cdc25C could be stably overexpressed in DC2.4 cells by LV-Cdc25C infection.DC2.4 cells transfected with LV-Cdc25C secreted more IL-6,IL-12,TNF-αand IFN-γ,and had higher expression levels of CD40,CD86,CCR7 and MHC-II than unaltered DC2.4 cells.The elevated Cdc25C in dendritic cells also further increased the secretion of IL-2,granzyme B and perforin to elicit Cdc25C-specific CTLs,and induced the higher cytotoxicity in Hepa1-6 cell lines(P<0.05),but this had no effect on the target cells when MHC-I monoclonal antibodies were blocked.Conclusion DC2.4 cells transfected with LV-Cdc25C can induce specific CTLs,and result in a strong cellular immune response.The dendritic cells that overexpress Cdc25C may be useful for hepatoma immunotherapy.

<i>In Silico</i>Pharmacological Analysis of a Potent Anti-Hepatoma Compound of Mushroom Origin and Emerging Role as an Adjuvant Drug Lead

Mushrooms are well-known to possess a continuum of anticancer metabolites that are vital in the development of anticancer adjuvant drug leads based on natural products. Owing to the fact that conventional cancer therapeutic methods were failed to lessen mortality caused by cancer to the estimated level with occurrence of adverse side effects, anticancer agents isolated from natural mushroom sources unarguably make an experimental research area worth mass focus today. The current study was targeted on in vitro cytotoxicity and in silico predictive pharmacological analysis of a flavonoid compound isolated from Fulvifomes fastuosus mushroom. Targeted compound was isolated from the mushroom using different chromatographic methods and identified by NMR spectrometry and mass spectrometry. Cytotoxicity experiments were carried out using MTT assay and apoptotic cells were identified by ethidium bromide/acridine orange staining. The SwissADME tool, BOILED-Egg construction model and Swiss target protein prediction software have been used to perform in silico predictive pharmacological analysis. The isolated compound has been identified as 2-(3,4-dihydroxyphenyl)-6-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-5'-methylspiro[2H-furo[3,2-c]pyran-3,2'-furan]-3',4-dione by spectrometric methods. The result of MTT assay showed that 2-(3,4-dihydroxyphenyl)-6-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-5'-methylspiro[2H-furo[3,2-c]pyran-3,2'-furan]-3',4-dione has potent anticancer activity for hepatoma against Hep-G2 cell line (IC50 = 20.8 μg/ml) being less toxic to normal CC-1 epithelial cells (IC50 = 167.00 μM). The cells treated with compound ex-hibited apoptotic features such as cellular shrinkage, nuclear fragmentation and condensed cytoplasm. In summary, 2-(3,4-dihydroxyphenyl)-6-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-5'-methylspiro[2H-furo[3,2-c]pyran-3,2'-furan]-3',4-dione has shown potent anticancer properties against hepatoma with less cytotoxicity effect on normal cells. Furthermore, in silico study has revealed that properties of 2-(3,4-dihydroxyphenyl)-6-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-5'-methylspiro[2H-furo[3,2-c]pyran-3,2'-furan]-3',4-dione may contribute to making a high absorption and clearance of the test compound as not interfering with the therapeutic failure of the compound. The properties of 2-(3,4-dihydroxyphenyl)-6-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-5'-methylspiro[2H-furo-[3,2-c]pyran-3,2'-furan]-3',4-dione were compatible with well-known anticancer drug lapatinib. In conclusion, 2-(3,4-dihydroxyphenyl)-6-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-5'-methylspiro[2H-furo[3,2-c]pyran-3,2'-furan]-3',4-dione has a high tendency to act as a good anticancer adjuvant drug in the treatment of hepatoma.

IN VITRO ANTI-HEPATOMA EFFECTS OF MONOCYTES AND KUPFFER CELLS ISOLATED FROM HEPATOMA PATIENTS AFTER TREATMENT WITH BIOLOGICAL IMMUNE STIMULANTS

Monocytes (MC), lymphocytes (LC) and Kupffer cells (KC) were isolated respectively from blood and surgical liver samples of patients suffering from he-patocellular carcinoma (HCC). 13 patients were given BCG, mixed bacterium vaccine (MBV) and human white blood cell interferon (IFN), the other 3 patients were not treated with any biological immune stimulants (BIS) and served as controls. The cytosta-tic and cytotoxic effects of MC and KC on human hepatoma SMMC-7721 (TC) were assayed in vitro and the numbers of T total (Tt), T helper (Th) and T suppressor (Ts) cells were counted using CD monoclonal antibody immunofluorescence. The results were as follows: (1) On the 7th day after the first administration of BIS, the cytostatic and cytotoxic effects of MC on TC showed obvious increase over pre-administration. The activity of BIS was 1 ?5 times as high as that in the controls. (2) After 3 administrations, the cytostatic effect of MC on TC increased to the normal level (84%), while the controls remained as before (45%). (3) On the 7th day after first administration, cytostatic and cytotoxic effects of KC on TC were 0.5 and 1 times higher respectively than those of the controls. (4) The numbers of Tt and Th of patients given BIS increased continuously; on the contrary Ts decreased in number. These results indicate that combined use of BCG, MBV and IFN can actively enhance the immune anti-hepatoma function of patients suffering from HCC.

基于靶点网络与分子对接探讨苦参介导NF-κB通路抗肝癌的作用机制

目的:运用网络药理学及分子对接的方法,探讨苦参Sophora flavescen Ait.中有效活性成分抗肝癌(hepatocellular carcinoma,HCC)的作用机制,并通过体内、体外实验对苦参抗肝癌的作用机制进行探究。方法:通过TCMSP、Swiss Target Prediction数据库和Genecards数据库和OMIM数据库等搜集苦参的有效活性成分和治疗HCC的潜在靶点;通过STRING数据平台构建靶点基因PPI网络,将degree值排序后筛选关键靶点;通过DAVID数据平台选取关键靶点的基因进行GO和KEGG通路富集分析;通过Discovery Studio筛选后选取CASP3、STAT3作为关键靶点与前5个活性成分进行分子对接;并运用体内、体外对应性实验对网络药理学所预测的结果进行苦参抗肝癌的药效学验证。结果:网络药理学结果筛选得到39个活性成分,涉及46个关键靶点,根据度值筛选,发现前20个基因主要富集在“癌症通路”等相关通路上,分子对接试验证明关键靶点NF-κBp65、CASP3、STAT3与有效活性成分之间存在较好的亲和力。苦参抗肝癌的实验结果显示,通过抑制NF-κBp65及其相关因子TNF-α、IL-6促进肝癌细胞凋亡。结论:该研究表明苦参可通过多种成分、不同靶点激活多条通路来发挥对肝癌细胞的拮抗机制,为阐明苦参抗肝癌的分子机制提供药理学数据参考。

天名精内生真菌的分离鉴定及抗肝癌活性研究

目的对天名精内生真菌进行分离和鉴定,并从中筛选出具有抗肝癌作用的活性菌株,为寻找新的抗肝癌先导化合物提供菌株资源。方法采用植物组织切块法和平板划线法对天名精内生真菌进行分离纯化;MTT法筛选对肝癌HepG2细胞增殖具有抑制作用的活性菌株,并计算活性显著内生真菌的IC50值;运用形态学鉴定和分子鉴定的方法对活性菌株进行鉴定。结果从天名精中共分离得到60株内生真菌,其中TMJ-03、TMJ-13、TMJ-15、TMJ-23、TMJ-54为活性内生真菌,其发酵物抑制HepG2细胞增殖的IC50值分别为19.49(TMJ-C-03)、31.13(TMJ-C-13)、88.65(TMJ-C-15)、37.42(TMJ-D-23)、17.22(TMJ-D-54)、32.72(TMJ-C-54)μg·mL^(-1)。5株活性菌株均鉴定到种,分别为Aspergillus terreus、Chaetomium globosum、Alternaria tillandsiae、Fusarium proliferatum、Alternaria arborescens。结论首次从天名精中筛选出对HepG2细胞增殖具有抑制作用的活性内生真菌,为寻找新的抗肝癌药物提供了菌株资源。

Anti-inflammatory effect and antihepatoma mechanism of carrimycin

BACKGROUND New drugs are urgently needed for the treatment of liver cancer, a feat that could be feasibly accomplished by finding new therapeutic purposes for marketed drugs to save time and costs. As a new class of national anti-infective drugs, carrimycin(CAM) has strong activity against gram-positive bacteria and no cross resistance with similar drugs. Studies have shown that the components of CAM have anticancer effects.AIM To obtain a deeper understanding of CAM, its distribution, metabolism and antiinflammatory effects were assessed in the organs of mice, and its mechanism of action against liver cancer was predicted by a network pharmacology method.METHODS In this paper, the content of isovaleryl spiramycin Ⅲ was used as an index to assess the distribution and metabolism of CAM and its effect on inflammatory factors in various mouse tissues and organs. Reverse molecular docking technology was utilized to determine the target of CAM, identify each target protein based on disease type, and establish a target protein-disease type network to ascertain the effect of CAM in liver cancer. Then, the key action targets of CAM in liver cancer were screened by a network pharmacology method, and the core targets were verified by molecular docking and visual analyses.RESULTS The maximum CAM concentration was reached in the liver, kidney, lung and spleen 2.5 h after intragastric administration. In the intestine, the maximum drug concentration was reached 0.5 h after administration. In addition, CAM significantly reduced the interleukin-4(IL-4) levels in the lung and kidney and especially the liver and spleen;moreover, CAM significantly reduced the IL-1β levels in the spleen, liver, and kidney and particularly the small intestine and lung. CAM is predicted to regulate related pathways by acting on many targets,such as albumin, estrogen receptor 1, epidermal growth factor receptor and caspase 3, to treat cancer, inflammation and other diseases.CONCLUSION We determined that CAM inhibited inflammation. We also predicted the complex multitargeted effects of CAM that involve multiple pathways and the diversity of these effects in the treatment of liver cancer, which provides a basis and direction for further clinical research.

藤茶总黄酮肠道代谢产物及其体外抗肝癌活性的研究

研究藤茶总黄酮口服给药后大鼠肠道代谢的产物及其体外抗肝癌活性。采用超高效液相色谱-高分辨质谱(UPLC-HRMS)联用技术分析总黄酮在大鼠肠道中的代谢产物,并以MTT法检测代谢产物对肝癌细胞HepG2增殖的抑制活性。通过UPLC-HRMS测定,总黄酮在大鼠粪便中检测到6个代谢产物。其主成分二氢杨梅素主要通过硫酸化、去羟基化、甲基化、还原去羟基化等反应发生代谢转化;其他成分杨梅苷、杨梅素则分别发生水解、去羟基化反应生成相应的代谢产物;这些代谢产物对HepG2细胞增殖有较好抑制作用,在不同作用时间点(48 h、72 h),其IC50分别是179.58、103.24μg/mL。藤茶总黄酮口服后在大鼠肠道中可发生多种代谢反应,其代谢产物具有体外抗肝癌活性。

阿可拉定长循环脂质体的制备与药效学评价

目的制备阿可拉定长循环脂质体(icaritin PEGylated liposomes,Ica-Lips),并评价其对人源肝癌细胞(hepaloblasloma G2,HepG2)的体内抗肿瘤效果。方法采用冷冻干燥-水化法制备Ica-Lips,并在透射电镜下观察Ica-Lips的微观形态,测定Ica-Lips的包封率、粒径分布、Zeta电位,以及在pH7.4磷酸盐缓冲液(PBS)以及大鼠血浆中的药物释放特性;并考察了Ica-Lips的稳定性;比较了Ica-Lips与Ica原料药在大鼠体内的药动学行为,评价Ica-Lips对小鼠接种人源肝癌细胞(HepG2)的抑瘤效果。结果制备的Ica-Lips呈类球状分布,其包封率为96.4%±0.3%,平均粒径为(108.4±3.6)nm,Zeta电位为(-12.9±0.2)mV;与Ica-Lips在pH7.4 PBS中的释药速率相比,其在大鼠血浆中的释药速率明显加快;Ica-Lips在低温条件下保存3个月,稳定性良好;药动学研究表明,Ica-Lips可显著延长药物在大鼠体内的滞留时间,增加药物的生物利用度;药效学研究表明,Ica-Lips对小鼠接种人源肝癌细胞(HepG2)的抑瘤效果显著高于Ica溶液(P<0.05)。结论将阿可拉定制备成长循环脂质体,提高了抗肿瘤效果,具有潜在的临床应用价值。

肉苁蓉总苷对HepG2细胞增殖、凋亡及Wnt/β-Catenin通路相关蛋白表达的影响

为了探讨肉苁蓉总苷(total glycosides of Cistanche deserticola,TG)对HepG2细胞的抑制作用及其机制研究。采用不同浓度(0、3.5、10.5、21、31.5、42μg/mL)TG处理HepG2细胞24 h后,使用CCK8法检测细胞存活率;应用Hoechst 33342/PI双染法及Annexin V-FITC/PI检测细胞凋亡;并通过细胞迁移试验检测细胞迁移现象;同时采用流式细胞仪检测细胞周期进展变化;通过Western blot法测定α-甲胎蛋白(α-fetoprotein,AFP)、β-连环蛋白(β-catenin)、蓬乱蛋白(Dishevelled,Dsh)、糖原合成酶激酶-3β(GSK-3β)的表达量。结果发现,TG可以降低HepG2细胞的增殖能力,且有浓度依赖性,当TG为42μg/mL时,细胞存活率仅有31.04%;此外,TG可以破坏细胞结构,诱导细胞凋亡,AV/PI检测后细胞凋亡率可高达32.44%;还可促使细胞坏死,限制细胞迁移,处理后的组别与对照组之间存在显著差异(P<0.05或P<0.01);同时,高浓度TG可以阻滞HepG2细胞在S期和G2/M期;与对照组相比,TG处理之后,β-catenin、Dsh表达下降,而GSK-3β表达量升高。由此可知,肉苁蓉总苷通过影响细胞周期进展、促进细胞凋亡、限制细胞迁移等方面抑制HepG2细胞增殖,其作用机制可能是通过Wnt/β-catenin信号通路,激活GSK-3β降解β-catenin来实现肝癌抑制作用的。

3,4-二氢萘-1(2H)-酮衍生物的合成及其生物活性评价

据报道,3,4-二氢萘-1(2H)-酮类化合物具有潜在的抗肿瘤活性和应用价值。以间苯二甲醛、7-甲氧基-3,4-二氢萘-1(2H)-酮、7-氟-3,4-二氢萘-1(2H)-酮为原料,以20%的氢氧化钠溶液为催化剂,通过Claisen-Schmidt缩合反应得到两个苯基桥连的3,4-二氢萘-1(2H)-酮类化合物1和化合物2,并通过NMR、IR、HRMS等波谱手段进行结构表征。MTT法评价了两个化合物对肝癌细胞HepG2、SMMC-7721、QGY-7703和正常肝细胞HHL-5的细胞毒性,ELISA法评价了化合物对LPS诱导的RAW264.7细胞中炎症细胞因子TNF-α、IL-6释放的抑制作用。与阳性对照药相比,化合物1和2显示了较好的抗肝癌活性,特别是氟取代的化合物2对HepG2的IC50值达到2.02μmol/L,对正常细胞HHL-5的毒性较小,且能有效抑制LPS诱导的RAW264.7细胞中炎症细胞因子TNF-α、IL-6的释放,表现出较好的抗炎活性。该研究为开发高效低毒的抗肝癌药物提供了有力素材。

负载紫檀芪的岩藻多糖-小麦醇溶蛋白纳米颗粒的制备及体外抗肝癌活性研究

紫檀芪是一种具有多种生物活性的疏水性天然产物,因低溶解度和光不稳定性而限制了其在实际中的应用。通过pH驱动法制备了具岩藻多糖涂层的小麦醇溶蛋白纳米颗粒,确定了小麦醇溶蛋白与岩藻多糖的最优质量比为8∶1,以该纳米颗粒为载体包封了紫檀芪,确定了紫檀芪与小麦醇溶蛋白的最优质量比为1∶10,最终制得的紫檀芪-小麦醇溶蛋白-岩藻多糖纳米颗粒呈近似球形,粒径为(122.34±1.08)nm,PDI为0.188±0.001,Zeta电位为(-27.73±0.44)mV,包封率为(94.09±1.60)%,载药率为(8.36±0.14)%,在pH为3.0~8.0的溶液和NaCl浓度为0~20 mmol/L的溶液中具良好的稳定性,长期储存稳定性良好,光照稳定性显著强于游离的紫檀芪;红外实验表明小麦醇溶蛋白、紫檀芪、岩藻多糖三者之间主要通过静电吸引、氢键和疏水作用结合。HepG2细胞的MTT实验和凋亡实验表明,与游离紫檀芪相比,紫檀芪-小麦醇溶蛋白-岩藻多糖纳米颗粒具有显著提高的体外抗肝癌活性。

功劳木的化学成分研究

目的研究功劳木的化学成分,评价功劳木化学成分体外抗肝癌HepG2细胞活性。方法功劳木经70%乙醇提取后经硅胶干柱色谱分成8份,第2～5份经硅胶柱色谱、葡聚糖凝胶Sephadex LH-20、大孔吸附树脂等技术分离纯化化合物,利用核磁共振技术鉴定它们的化学结构,并通过测试体外最低抑制浓度评价有关物质的抗肝肿瘤活性。结果分离、纯化并鉴定了8个化合物,分别为erythro-syringoylglycerol 8-O-β-D-glucoside(1)、3,4,5-三甲氧基苯酚1-O-β-D-葡萄糖苷(3,4,5-trimethoxyphenyl-1-O-β-D-glucoside,2)、表丁香脂素(episyringaresinol,3)、5,5'-二甲氧基落叶松脂醇4'-O-β-D-葡萄糖苷(5,5'-dimethoxylariciresinol-4'-O-β-D-glucoside,4)、β-谷甾醇(β-sitosterol,5)、小檗碱(berberine,6)、巴马汀(palmatine,7)、药根碱(jatrorrhizine,8)。结论化合物1～4皆为首次从该属植物中分离得到。化合物3、6、8显示出不同程度的抑制肝癌HepG2细胞活性。

鹰嘴豆肽抑制肿瘤作用和对免疫功能的影响

目的:探讨鹰嘴豆肽抑制肝癌H22细胞移植瘤的效果及对免疫功能的影响。方法:将小鼠随机分为正常对照组、阳性对照组、肿瘤对照组和鹰嘴豆肽低、中、高剂量组(50、100、200mg/(kg·d)),除正常组外,其余各组小鼠均在皮下接种肝癌H22细胞,建立肝癌小鼠模型。建模后第2天开始灌胃给药,连续给药10d。末次给药24h后拉托颈椎处死小鼠,测体质量、肿瘤大小、抑瘤率、胸腺指数、脾指数、巨噬细胞吞噬能力、脾细胞吞噬能力等。结果:鹰嘴豆肽低、中、高3个剂量均可抑制H22肿瘤的生长,且没有阻碍小鼠体质量和免疫器官的增长。与肿瘤对照组相比,鹰嘴豆高剂量组可以显著提高小鼠迟发型超敏反应(DTH)的能力、淋巴细胞和巨噬细胞吞噬能力(P<0.05),鹰嘴豆低、中剂量组对其也有一定的改善作用,但是没有表现出剂量依赖效应。讨论:鹰嘴豆肽可能是通过增强H22肝癌小鼠的免疫功能来抑制肿瘤的生长,具体的作用机制还需要更深入的研究。

药物筛选微流控芯片的构建及其在花蕊石抗肝癌研究中的应用

目的构建一种集成有微阀结构的双层PDMS-玻璃复合芯片,考察其适用性并研究花蕊石生品、制品水煎液对肝肿瘤细胞Hep G2凋亡坏死的影响。方法制作双层微流控芯片并考察微阀的性能,同时进行了Hep G2细胞的培养,死活试剂盒检测细胞活力。将预制备的不同浓度花蕊石生品、制品水煎液通入微流控芯片,分别作用于Hep G2细胞24、48和72 h,凋亡坏死试剂盒检测Hep G2细胞凋亡坏死率。结果芯片微阀性能良好,可灵活控制液阀的开关且未表现出明显的延迟现象。细胞在芯片中生长状态良好,培养72 h期间细胞存活率≥97%。药物作用细胞后,发生凋亡坏死的细胞数量随给药浓度的升高、给药时间的延长而增加,呈现出一定的时间与浓度依赖性,且制品水煎液的效果好于生品。结论实验证明了花蕊石生品、制品水煎液对肝肿瘤细胞Hep G2具有促凋亡作用,也进一步验证了本实验室设计的芯片在细胞研究和药物筛选方面的可行性,为微流控芯片在中药抗肿瘤新药开发领域中的推广与应用提供理论依据。

福建绞股蓝皂苷的优化提取及其抗肝癌活性

绞股蓝素有"南方人参"美誉,含有与人参皂苷相同骨架达玛烷型结构的三萜皂苷。通过不同体积分数乙醇溶液优化提取绞股蓝皂苷,测定其抗癌活性,为后续活性化合物的分离奠定基础。利用HPLC分析绞股蓝的皂苷成分并确定提取绞股蓝皂苷的乙醇体积分数,正交实验优化提取方法,进而MTT法测定提取的绞股蓝皂苷抗肝癌活性。HPLC分析确定用体积分数75%乙醇溶液提取绞股蓝总皂苷(GYP),体积分数45%乙醇提取极性绞股蓝皂苷(GYP1),体积分数90%乙醇提取弱极性绞股蓝皂苷(GYP2);正交实验结果表明在70℃水浴中以30质量的体积分数45%乙醇加热提取12 h为最佳条件提取GYP1,在60℃水浴中以30质量的体积分数90%乙醇加热提取6 h为最佳条件提取GYP2;利用MTT法测定GYP、GYP1和GYP2对肝癌细胞的抑制作用,GYP和GYP2对肝癌细胞SMCC7721具有抗癌活性,半抑制率IC50值分别为(165.83±14.12)μg/mL和(113.97±9.26)μg/mL,GYP1没有抗癌活性,表明绞股蓝抗癌药效成分主要是由弱极性绞股蓝皂苷发挥功效。

单抗导向阿霉素免疫毫微粒抗肝癌作用的机制

目的 研究抗人肝癌单克隆抗体HAb18为导向载体的阿霉素 (ADR )人体白蛋白 (HSA)免疫毫微粒HAb18 ADR HSA NP抗肝癌作用的机制。方法 利用激光共聚焦仪和透射电镜观察HAb18 ADR HSA NP在人肝癌细胞SMMC 772 1中的内化作用 ,通过扫描电镜和透射电镜观察HAb18 ADR HSA NP对SMMC 772 1多药耐药株 (SMMC 772 1/MDR+ )的结合和内化现象 ,采用四甲基偶氮唑蓝比色法测定HAb18 ADR HSA NP对SMMC 772 1及其耐药细胞的杀伤作用。结果 HAb18 ADR HSA NP在SMMC 772 1细胞中存在内化现象 ,且该内化与温度有关 ,具抗体特异性。同时 ,HAb18 ADR HSA NP在SMMC 772 1/MDR+ 表面结合 ,并能内化 ;且其能增强SMMC 772 1/MDR+ 对ADR杀伤的敏感性。结论 HAb18 ADR HSA

100种常用藏药抗肝癌的体外活性筛选

为从藏药中发现新的抗癌药物,从青海互助县收集了100种常用藏药,采用快速溶剂萃取法制备出了各种藏药的环己烷、乙酸乙酯和甲醇提取物.选用2种肝癌细胞株Hep G2和SMMC-7721,以MTT法测试了100种藏药的抗肝癌活性,选用人肝细胞株LO-2评价了藏药的体外毒性.结果表明:15种藏药材显示出抗肝癌活性,IC50值均小于150μg/m L,其中藜、天名精、三脉紫菀3种藏药的乙酸乙酯提取物显示出显著的抗肝癌活性,其IC50值均小于50μg/m L,而对正常肝细胞LO-2未显示出明显的毒性.该研究为进一步从藏药中发现新的抗癌药物提供了思路.

佛甲草中细胞毒活性成分的研究

采用中压柱色谱和制备高效液相色谱法对中国传统民族药佛甲草(Sedum lineare Thunb)的甲醇提取物化学成分进行分离,从中分离得到6个化合物.首次对化合物山奈酚7-O-6″-O-丙二酸单酰-D-吡喃葡萄糖苷(1)的波谱数据进行了归属,用MTT法测试6个单体成分对人肝癌细胞株HepG2和Hep3B的抑制作用.结果显示6个化合物均对HepG2和Hep3B细胞表现出抗肿瘤活性,并且IC50值在22.6~91.5μg/mL之间,在浓度大于100μg/mL时也没有表现出对人正常肝细胞L-02的抑制作用.相比于黄酮类化合物1、2,苯丙酸类化合物3~6(IC50=22.6~44.3μg/mL)对HepG2和Hep3B细胞表现出更强的抑制作用.其中,顺式苯丙酸类化合物5对HepG2和Hep3B细胞的抑制作用最明显.其IC50值范围在22.6~28.2μg/mL,能明显抑制人肝癌细胞的增殖,诱导其凋亡.

特异性抗肝癌单味中草药提取物的高通量筛选

目的利用高通量筛选分析技术评价单味中草药提取物对肝癌细胞及成纤维细胞的作用,以期获得具有特异性抗肝癌作用的中草药提取物。方法将242种常用单味中草药分别应用石油醚、乙醇或水进行提取,共获得554个中草药提取物。采用MTT来评价提取物对人肝癌细胞Bel7402和小鼠成纤维细胞NIH3T3存活的作用。结果提取物对人肝癌细胞Bel7402及成纤维细胞NIH3T3生长抑制作用>50%的样品分别占总样品数的7.4%和14.8%,对2种细胞抑制率同时>50%的样品占总数的4.4%,对Bel7402抑制率为对NIH3T3抑制率2倍以上且对Bel7402抑制率>50%的样品占总样品数的1.6%。复筛结果显示,防己乙醇提取物(DF173)对肝癌细胞的细胞毒作用特异性较好,具有良好的量效关系。DF173对Bel7402细胞毒作用IC50为8.27 mg·L-1,对NIH3T3的细胞毒作用IC50为19.48 mg·L-1。结论肿瘤细胞联合正常成纤维细胞筛选评价中草药提取物特异性抗肿瘤作用,有利于发现高效、低毒抗肿瘤中草药提取物。