小鼠H_(22)肿瘤的酶组织化学和超微结构观察

光镜下H_(22)肿瘤细胞弥漫索样排列,网状纤维包绕瘤细胞团形成巢状,胶原纤维极少。碱性磷酸酶反应产物主要附着于细胞膜部位;酸性磷酸酶细胞内散在分布;细胞色素氧化酶及乳酸脱氢酶弥散位于细胞浆内。各种酶反应产物均较匀细。扫描和透射电镜示瘤细胞表面微绒毛发达,细胞间可见桥粒连接,线粒体、粗面内质网和多聚核糖体丰富,胞浆内细胞器有极性分布倾向。以上结果不仅为该瘤株的进一步应用提供了有益的参考资料,同时提示关于H_(22)肿瘤已转变为多形细胞肉瘤的结论是值得商榷的,其应是一株低分化肝癌腹水瘤模型。     

Clinical Observation on Primary Hepatocarcinoma Treatedby Arterial Perfusion with Cinobufotalin and Chemicals

Thisarticlereportedthetreatmentof38patientsofprimaryhepatocarcinomawitharterialperfusionwithcinobufotalinandchemicals,whichgainedsatisfactoryefficacy.METHODSClinicalMaterialAccordingtothestandardofdiagnosis,typing,staging,andefficacyevaluationforhepatocarcinomaformulatedinDec.1977(1),allthe70patientsenrolledforobservationwereconfirmedtobeprimaryhepatocarcinomabyclinical,B--ultrasonic,CTandblooda--fetaprotein(AFP)examination,withtheirhepatitisBsurfaceantigen(HBsAg)shownaspositive.Thepat…     

甲乙煎影响H_(22)小鼠肝癌组织黏附分子E-cadherin表达的实验研究

目的:探讨甲乙煎对肝癌组织中细胞黏附分子E-cadherin表达的影响.方法:复制H22小鼠肝癌淋巴道转移模型,用中药甲乙煎水煎液(大、中、小剂量)、生理盐水灌胃和5-Fu腹腔注射20天,计算脾指数、胸腺指数,通过免疫组化法观察局部瘤组织中细胞黏附分子E-cadherin的表达水平.结果:中药大、中、小剂量组的脾指数、胸腺指数均高于生理盐水和5-Fu组,中药大、中、小剂量组和5-Fu组E-cadherin表达均高于生理盐水组.结论:甲乙煎可改善小鼠的免疫功能,提高H22小鼠肝癌局部瘤组织中细胞黏附分子E-钙黏蛋白的表达水平.     

青蒿醇提物对H22肝癌小鼠的抑瘤作用研究

目的：观察青蒿醇提物对H：：肝癌小鼠的体内抑瘤作用。方法：取H。肝癌细胞接种于小鼠右腋皮下,随机分为模型对照组、5-氟尿嘧啶组（5-Fu）及青蒿醇提物125mg／（kg·d）组、500mg／（kg·d）组,5-Fu组肌肉注射给药,其余各组灌胃给药。连续给药8d后,剖瘤称取质量,计算抑瘤率;利用高效液相色谱仪测定青蒿醇提物中青蒿素含量。结果：与模型对照组比较,青蒿醇提物各剂量组均不同程度的抑制肿瘤生长（P〈0．Ot）,青蒿醇提物125me,／（kg·d）组和青蒿醇提物500m/（kg．d）组的抑瘤率分别为54．6％、50．6％;本实验青蒿醇提物中青蒿素含量为0．02％,而在青蒿素抗H。小鼠肝癌试验中,青蒿素在剂量为10mg／（kg·d）时,对H22小鼠肝癌已没有明显的抑制作用。结论：青蒿醇提物对肝癌有明显的抑制作用;除青蒿素以外,中药青蒿中可能存在其它的抗肿瘤成分。     

β-榄香烯对人肝癌细胞侵袭转移及相关机制的实验研究

目的研究β-榄香烯对人肝癌细胞侵袭转移能力的影响,并探讨其可能的作用机制。方法应用不同浓度的β-榄香烯作用于人肝癌细胞株HepG2 24h,以MTT法检测细胞黏附能力,采用Matrigel试剂、Transwell实验分别检测细胞迁移和侵袭能力,Western blot法检测细胞基质金属蛋白酶MMP-2及MMP-9蛋白的表达。结果随β-榄香烯浓度增大细胞黏附能力降低,同时侵袭及迁移细胞数目明显减少;β-榄香烯可以浓度依赖性地减少MMP-2及MMP-9蛋白表达。结论β-榄香烯可有效抑制人肝癌细胞侵袭转移能力,这主要是通过减少MMP-2及MMP-9蛋白的表达途径来实现。     

甘草次酸类化合物的制备和抗肿瘤活性研究

目的制备18α-和18β-甘草次酸类复合物,并对体外抗癌活性进行研究。方法以18β-甘草次酸(Ⅰ)为起点,进行构型转化得到其光学异构体18α-甘草次酸(Ⅱ),再经C30-位羧基的甲酯化分别得到18β-甘草次酸甲酯(Ⅲ)和18-α-甘草次酸甲酯(Ⅳ),并与环磷酰胺的抗癌活性体内代谢产物——二氯磷酰氮芥偶联而得2个抗癌复合物18β-甘甲磷氮芥(Ⅴ)和18α-甘甲磷氮芥(Ⅵ);用四大光谱法(NMR、MS、IR、UV)对各化合物的化学结构进行鉴定分析;利用人类肝癌细胞株BEL-7402为体外实验模型,用MTT法观察各化合物对人肝癌细胞增殖的抑制活性;抗肿瘤药物顺铂作为阳性对照物。结果对化合物的合成工艺进行优化;18α-甘草次酸的构象转化得率为45%;目标化合物18β-甘甲磷氮芥(Ⅴ)和18α-甘甲磷氮芥(Ⅵ)的产率分别为35%和25%。所制备的甘草次酸衍生物中,化合物Ⅱ和Ⅵ对BEL-7402肿瘤细胞的增殖显示明显强的抑制活性,浓度在5～500μg/mL范围内,对肿瘤细胞增殖的抑制率分别为2.6%～86%及1.3%～50.1%;在相同条件下,对照物顺铂的抑制率为20.0%～73.41%。结论目标化合物的制备中磷酰酯化反应的条件控制(无水、物料比1∶1.25,温度65℃和反应时间3～4h)是合成关键;化合物Ⅱ和Ⅵ在中高浓度时,与顺铂具有较类似的抑制肝癌细胞增殖活性。甘草次酸类化合物的构型转化及与抗癌基团偶联可能提高其抗癌潜力。     

Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway

Objective:To study the immunologic function of dendritic cells(DCs) cultured in two kinds of hepatoma cell line’s supernatant and the enhancing effects of carboxymethylpachymaran(CMP) on DCs.Methods:DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin(IL)-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines(HepG2 and HepG2.2.15) were collected for condition medium(CM) according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3:1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens(HLA-DR) were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction(MLR) stimulated by DCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide(MTT) analysis.IL-12p70,interferon-γ(IFN-γ),and nuclear factor k B(NF- k B) were detected by enzyme-linked immunosorbent assay analysis.Results:The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group(P<0.01).Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group(P<0.05).Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups(P<0.05).After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added(P<0.05).However,there was no significant difference between the two CM groups(P>0.05).Conclusions:Two kinds of hepatoma cell line’s supernatant can inhibit the immunologic function of DCs.This suppressive effect may be related to the inhibition of NF-κB/Rel pathway.CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.     

蜂毒素对人肝癌细胞中HMGB1及VEGF-C表达的影响

目的观察蜂毒素对人肝癌细胞株HepG2中高迁移率族蛋白B1(high mobility group box1,HMGB1)及血管内皮生长因子C(vascular endothelial growth factors C,VEGF-C)表达的影响,探讨其抑制肝癌细胞的作用机制。方法肝癌细胞HepG2体外培养,经蜂毒素处理后,采用四甲基偶氮唑蓝(MTT)法了解蜂毒素对肝癌细胞增殖的影响,用Western-blotting、qRT-PCR方法检测HMGB1、VEGF-C的表达。结果蜂毒素在体外能够抑制肝癌细胞的增殖活性;Western-blotting结果显示,蜂毒素可抑制HMGB1的表达,且呈浓度依赖性,但对VEGF-C的蛋白表达无明显影响;qRT-PCR结果显示,蜂毒素在mRNA水平均具有抑制HMGB1、VEGF-C表达的作用。结论蜂毒素可降低肝癌细胞的增殖活性,并可能通过HMGB1、VEGF-C的表达发挥抗肝癌作用。     

卡铂碳包铁纳米磁性微球对移植性肝癌热疗疗效机制的研究

目的 卡铂Fe@C纳米笼壳聚糖微球 (C-Fe@C-CN) 对移植性肝癌模型具有较好的热疗效果,探讨热疗对肿瘤细胞治疗的分子机制.方法 用制备好卡铂Ve@C纳米笼壳聚糖微球 (C-Fe@C-CN) ,对50只移植性肝癌大鼠模型 (随机分为5组,A组生理盐水组、B组卡铂组,C组微球组,微球热疗30min D组和微球热疗60min E组) ,经肝动脉注射给药24h后,免疫组织化学法观察热疗对肝癌肿瘤细胞HSP70表达的影响,流式细胞术检测对肝癌细胞凋亡率及Fas蛋白表达的影响.结果 HSP70蛋白表达,A、B、c组 (n=6) 均为阴性,E组明显高于D组[ (96.11%±15.40%) n=6) vs (74.71%±13.85%) (n=6) ,p<0.05].D、E组肝癌细胞凋亡率[ (58.37%±13.84%) 、 (64.87%±15.72%) ]和Fas蛋白表达[ (35.23%±15.72%) 、 (38.45%±5.64%) ]较其他组显著增高 (P<0.01) ,D组与E组比较差异无显著性 (P>0.05) .结论 C-Fe@C-CN具有良好的磁靶向力和感应产热性能,加热可以诱导HSP70蛋白的大量产生,可以激活Fas基因诱导肝癌细胞凋亡,是一种潜在的抗肿瘤磁性纳米靶向热疗药物栽体.