结肠癌患者血清可溶性sIL-2R含量的变化

目的探讨结肠癌患者手术前血清可溶性白介素２受体（ｓｏｌｕｂｌｅｉｎｔｅｒｌｕｋｉｎ２ｒｅｃｅｐｔｏｒ，ｓＩＬ２Ｒ）含量改变与疾病的诊断、分期、预后判断等的关系．方法采用ＥＬＩＳＡ法对２７例结肠癌患者（ＤｕｋｅｓＡ，Ｂ期１９例），Ｃ，Ｄ期８例）及４２例对照组（正常３２例及肠易激综合征１０例）分别测定血清ｓＩＬ２Ｒ含量．同时使用间接荧光染色法对各组进行血ＣＤ３，ＣＤ４，ＣＤ８及ＣＤ４／ＣＤ８的测定．结果结肠癌患者ＤｕｋｅｓＡ，Ｂ期组血清ｓＩＬ２Ｒ含量（８３５ｐｍｏｌ／Ｌ±２１８ｐｍｏｌ／Ｌ）与对照组（６９２ｐｍｏｌ／Ｌ±２７９ｐｍｏｌ／Ｌ及７６２ｐｍｏｌ／Ｌ±２４６ｐｍｏｌ／Ｌ）无明显差异，而有淋巴结转移的ＤｕｋｅｓＣ，Ｄ期患者（３２１６ｐｍｏｌ／Ｌ±２３４４ｐｍｏｌ／Ｌ）则明显高于ＤｕｋｅｓＡ，Ｂ期患者，且ＣＤ４／ＣＤ８值明显下降（１０６±０４９比１５８±０３６）；另外，血清ｓＩＬ２Ｒ含量还与肿瘤细胞分化程度密切关联．结论结肠癌术前血清ｓＩＬ２Ｒ含量与疾病分期及预后有关．     

血清CA-50含量对消化系肿瘤的诊断价值

目的评价血清ＣＡ＿５０对消化系肿瘤的诊断价值．方法消化系肿瘤患者１７２例，其中肝癌５４例，胃癌４３例，大肠癌５７例，胰腺癌１８例；消化系良性疾病患者８８例，其中肝硬变３６例，胃良性病变（胃炎、胃溃疡）５２例；正常对照者６０例．全部受测对象均抽空腹静脉血，分离血清，－２０℃贮存备测．采用ＲＩＡ法测定血清ＣＡ５０含量．放免药盒由中国医学科学院肿瘤研究所提供，使用国产ＦＪ６３０型γ计数仪．数据均用ｘ±ｓ表示，以正常ｘ＋２ｓ作为上限计算阳性率．结果肝癌、胃癌、胰腺癌和大肠癌血清ＣＡ５０含量分别为２４０ｋＵ／Ｌ±２１８ｋＵ／Ｌ，１２１ｋＵ／Ｌ±１０６ｋＵ／Ｌ，１８２ｋＵ／Ｌ±１０７ｋＵ／Ｌ和１６１ｋＵ／Ｌ±１１３ｋＵ／Ｌ．显著高于正常对照组及消化道良性病变组（分别为５６ｋＵ／Ｌ±４４ｋＵ／Ｌ和５６ｋＵ／Ｌ±２１ｋＵ／Ｌ，Ｐ＜００１）．消化道肿瘤有腹腔及远处转移者，其血清ＣＡ５０含量升高更为明显．结论ＣＡ５０是较好的肿瘤标记物，有助于诊断消化道肿瘤．     

顺铂诱导人胃癌SGC-7901细胞

目的探讨顺铂对胃癌（ＳＧＣ７９０１）细胞凋亡的诱导作用，以揭示顺铂治疗胃癌的作用机制．方法选用不同剂量顺铂与人胃癌细胞系ＳＧＣ７９０１共同孵育不同时间后，应用光镜、电镜进行细胞形态学观察；细胞ＤＮＡ经特异性荧光染色后，用流式细胞仪分析细胞周期变化．结果细胞经顺铂处理后浓缩、深染、致密的染色质沿核膜下凝集、有凋亡小体形成．细胞周期分析Ｇ１期峰前出现典型的细胞凋亡峰，以２５ｍｇ／Ｌ顺铂作用２４ｈ，５０ｍｇ／Ｌ作用１６ｈ为例，定量分析凋亡细胞发生率分别为９３％和１４９％．结论顺铂可通过诱导人胃癌细胞系ＳＧＣ７９０１发生细胞凋亡达到消灭肿瘤细胞的目的．     

鼠自身免疫性肝炎模型的建立

目的探索鼠实验性自身免疫性肝炎（ＥＡＨ）模型的建立．方法用♀重１３０ｇ～１６０ｇＷｉｓｔａｒ大鼠分成８组，♀重２５ｇ～３５ｇＢＡＬＢ／ｃ小鼠分成２组（每组ｎ＝６）．Ⅰ组为ＰＢＳ对照组；Ⅱ组为ＣＦＡ；Ⅲ组为大鼠Ｓ１００２５ｍｇｉｐ；Ⅳ组为大鼠Ｓ１００２５ｍｇ加ＣＦＡｉｍ；Ⅴ组为大鼠Ｓ１００５ｍｇ加ＣＦＡｉｐ；Ⅵ组为大鼠Ｓ１００１０ｍｇ加ＣＦＡｉｐ；Ⅶ组为大鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ；Ⅷ组为小鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ组；Ⅸ组为大鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ给小鼠组；Ⅹ组为小鼠Ｓ１００２５ｍｇ加ＣＦＡｉｐ给小鼠组．观察生存率、肝功能、免疫球蛋白、肝脏的病理改变．结果用同种肝抗原Ｓ１００辅以福氏完全佐剂腹腔免疫Ｗｉｓｔａｒ大鼠可诱导ＥＡＨ模型，该法形成率高（９８％）、死亡率低（２％）．病理检查显示典型的ＥＡＨ病变．ｗｋ５血清ＡＬＴ为２００ＩＵ／Ｌ、ＩｇＧ为５５ｇ／Ｌ．结论Ｓ１００可以诱导典型的ＥＡＨ模型     

移植培养人肝细胞对急性肝衰竭小鼠的保护作用

目的探讨腹腔移植微载体粘附培养人肝细胞对Ｄ氨基半乳糖（ＤＧａｌ）诱导的急性肝衰竭小鼠的保护作用．方法采用胶原酶灌流法分离人肝细胞，胶原被覆的微载体Ｃｙｔｏｄｅｘ３加以培养，经腹腔注射２×１０６个肝细胞及载体，观察急性肝衰竭小鼠（ｎ＝３２，２５ｇ／ｋｇＤＧａｌ诱导４ｈ）７２ｈ存活率、血清ＡＬＴ、总胆红素以及肝脏病理变化．结果与只接受微载体而无粘附肝细胞的对照组比较，腹腔移植培养肝细胞对肝衰竭小鼠的存活率有明显改善（６５％ｖｓ０％），其血清ＡＬＴ（ＩＵ／Ｌ，２１７４４±２６３０ｖｓ４２６３１±４９２８，Ｐ＜００５）及总胆红素（ｍｍｏｌ／Ｌ，２６９１±３７６ｖｓ４１６８±３８３，Ｐ＜００５）较低．肝组织病理改变较轻．结论腹腔移植微载体粘附培养肝细胞可提供代谢支持，使ＤＧａｌ损伤的小鼠肝功能恢复．     

HBV转基因小鼠暴露ATB-1后肝脏ATB-1-DNA加成物水平的测定

目的探讨ＨＢＶ与ＡＴＢ１协同致肝癌的机理．方法用ＲＩＡ法测定ＨＢＶ转基因小鼠与正常小鼠暴露ＡＴＢ１后０，０５，１，２，４，８，２４ｈ７个不同时相肝脏ＡＴＢ１ＤＮＡ加成物的含量变化．结果暴露ＡＴＢ１后，ＨＢＶ转基因小鼠肝脏ＡＴＢ１ＤＮＡ加成物含量在各时相均高于正常小鼠，尤以１ｈ高峰时相值（５５９２ｐｍｏｌ／ｍｇ±４１５ｐｍｏｌ／ｍｇ比４１３６ｐｍｏｌ／ｍｇ±２８２ｐｍｏｌ／ｍｇＤＮＡ，Ｐ＜００１）及２４ｈ时相值（２４８７±２０３比９８９±８５，Ｐ＜００１）最显著．２４ｈ后，转基因小鼠肝脏ＡＴＢ１ＤＮＡ加成物仍维持高水平，但正常小鼠已基本恢复至暴露前水平．结论ＨＢＶ转基因小鼠暴露ＡＴＢ１后肝脏ＡＴＢ１ＤＮＡ加成物含量增加可能为ＨＢＶ与ＡＴＢ１协同致肝癌的直接原因．     

梗阻性黄疸内毒素血症与细胞免疫功能的关系

目的研究梗阻性黄疸免疫功能及其与内毒素血症的相关性．方法检测２８例梗阻性黄疸患者及２０例健康对照者血清内毒素，Ｔ淋巴细胞亚群及血清ＳＩＬ２Ｒ的水平．结果梗阻性黄疸患者血清内毒素和ＳＩＬ２Ｒ水平较对照组明显升高（４７０ｎｇ／Ｌ±１１３ｎｇ／Ｌ和７２５ｋＵ／Ｌ±２０１ｋＵ／Ｌｖｓ２８４ｎｇ／Ｌ±１０３ｎｇ／Ｌ和３２４ｋＵ／Ｌ±１１６ｋＵ／Ｌ，Ｐ＜００１），Ｔ淋巴细胞亚群ＣＤ３，ＣＤ４，ＣＤ４／ＣＤ８明显降低（５０４％±３３％和２９９％±３８％ｖｓ６３８％±４４％和３８３％±２８％，Ｐ＜００１；１２２±０３２ｖｓ１４３±０３７，Ｐ＜００５），同时亦发现梗阻性黄疸内毒素血症组较非内毒素血症组ＣＤ３，ＣＤ４水平明显减低，ＳＩＬ２Ｒ水平明显升高（４７４％±５１％和２７６％±５２％和８６７ｋＵ／Ｌ±２３１ｋＵ／Ｌｖｓ５２３％±５２％和３１２％±４３％和６７４ｋＵ／Ｌ±１８９ｋＵ／Ｌ，Ｐ＜００５）．相关分析显示血清内毒素水平与血清ＳＩＬ２Ｒ水平呈显著正相关（ｒ＝０８５１７，Ｐ＜００１）．结论梗阻性黄疸时内毒素血症与免疫功能状态密切相关．     

雷公藤多甙对大鼠急性肝衰竭的保护作用

目的探讨雷公藤多甙（ＧＴＷ）对实验性肝衰竭的保护作用．方法雄性Ｗｉｓｔａｒ大鼠２４只，随机分为对照组（ｎ＝８），急性肝衰竭（ＡＨＦ）组（ｎ＝８）和ＧＴＷ保护组（ｎ＝８）．ＧＴＷ组在实验前５ｄＧＴＷ２５ｍｇ／（ｋｇ·ｄ）经胃管灌胃，其余两组均以等量生理盐水溶液灌胃．从第６天开始，ＡＨＦ组和ＧＴＷ组均在空腹１２ｈ后ｉｐＤＧａｌＮ１６ｇ／ｋｇ，注射后４０ｈ，以３０ｇ／Ｌ戊巴比妥钠（４０ｍｇ／ｋｇ）ｉｐ麻醉，心脏抽血测定血清ＡＬＴ，ＴＢ和Ｔ淋巴细胞亚群．同时，在光镜和电镜下观察各组肝组织病理变化．结果ＡＨＦ组ＡＬＴ（ＩＵ／Ｌ）和ＴＢ（μｍｏｌ／Ｌ）分别为７８２８±５７６２和１２５６２７±６７０２７；ＯＸ８水平为１４０％±３％．ＧＴＷ组ＡＬＴ和ＴＢ分别为３５９±５４和４７±３５；ＯＸ８为４３％±４％．两组比较有显著统计学差异（Ｐ＜００５或００１）．同时，前者电镜下线粒体和内质网肿胀、破损，核内染色质凝聚，后者细胞器受损明显减轻．结论雷公藤多甙对实验性急性肝衰竭具有保护作用．     

胃癌及消化性溃疡患者胃窦粘膜胃肠激素的变化

目的探讨胃癌及消化性溃疡（ＰＵ）患者胃窦粘膜胃肠激素变化的意义．方法内镜及活检确诊的浅表性胃炎（ＣＳＧ）１０例，胃溃疡（ＧＵ）１５例，十二指肠溃疡（ＤＵ）１２例，胃癌（ＧＣ）６例．胃镜下取胃窦粘膜，用ＲＩＡ法测定胃泌素（Ｇａｓ）、生长抑素（ＳＳ）、Ｐ物质（ＳＰ）的含量，各组间进行比较．结果胃窦粘膜ＳＳ含量在ＧＵ，ＤＵ，ＣＳＧ，ＧＣ组分别为２５１ｐｇ／ｍｇ±１９４ｐｇ／ｍｇ（以下同），４７０±１７９，５３２±２１１及１２９３±５２３。其中ＧＵ组低于其余各组（Ｐ＜００５），而ＧＣ时则显著升高（Ｐ＜００１）．ＳＰ含量在ＤＵ组显著降低，与ＧＵ，ＣＳＧ，ＧＣ比较分别为４７９±１５７ｖｓ７６５±４１５，７８９±３９０及８０１±３４６，Ｐ＜００５；ＧＣ患者Ｇａｓ水平显著高于ＣＳＧ组，为４６４５±２９４４，ｖｓ２７６８±１５７２，Ｐ＜００１．结论胃粘膜中Ｇａｓ，ＳＳ，ＳＰ含量的变化可能在ＰＵ及胃癌的发病机理中起重要作用．     

褐藻胶对大鼠实验性肝纤维化的防治作用

目的研究褐藻胶对大鼠实验性肝纤维化的防治作用．方法用４０％四氯化碳（ＣＣｌ４）制备大鼠肝纤维化模型，实验分组为正常对照组（ｎ＝８），ＣＣｌ４组（ｎ＝８），秋水仙硷（ＣＯＬ）组（ｎ＝６）和褐藻胶组（ｎ＝６）（２００ｍｇ／ｋｇ，ｉｇ，３次／周×４）．观察肝脏组织学和血清Ⅲ型前胶原（ＰＣⅢ）及透明质酸（ＨＡ）水平的变化．结果褐藻胶组ＰＣⅢ（１４２８μｇ／Ｌ±７６１μｇ／Ｌ）和ＨＡ水平（２６５５μｇ／Ｌ±９３１μｇ／Ｌ）显著低于ＣＣｌ４组（２９３５μｇ／Ｌ±７８３μｇ／Ｌ，５１９８μｇ／Ｌ±１１８３μｇ／Ｌ，Ｐ＜００１），而褐藻胶组与ＣＯＬ组间无显著差异．病理学观察显示，ＣＣｌ４组肝纤维化程度重于褐藻胶组和ＣＯＬ组．结论褐藻胶对实验性肝纤维化有防治作用．     

七叶皂苷钠抑制人甲状腺未分化癌HTh74细胞株增殖作用的研究

目的 研究七叶皂苷钠对人甲状腺未分化癌HTh74细胞株增殖的潜在抑制作用及其分子机制.方法 以不同浓度的七叶皂苷钠(0,20,40,60,80 μmol/L)处理HTh74细胞48 h后,观察细胞形态变化;以不同浓度的七叶皂苷钠(0,20,40,60,80 μmol/L)处理HTh74细胞24h、48 h和72 h后,噻唑蓝比色(MTT)法检测七叶皂苷钠对HTh74细胞株增殖的抑制效应并计算细胞增殖率及半数抑制率(IC50);以不同浓度的七叶皂苷钠(0,5,10,15,20,25 μmol/L)处理HTh74细胞48 h后,Western印迹检测七叶皂苷钠引起的蛋白激酶B(Akt)、磷酸化Akt(p-Akt)以及细胞周期相关蛋白的表达.结果 七叶皂苷钠作用后,细胞发生明显皱缩,且作用浓度越高细胞皱缩越明显;MTT法结果显示,七叶皂苷钠对HTh74细胞有显著的生长抑制作用,呈现典型的时间与剂量依赖性,80 μmol/L七叶皂苷钠作用24 h,对细胞的生长抑制作用有统计学意义(F=111.4,P＜0.01),而40 μmol/L作用48 h及72 h时,细胞生长均受到抑制(F=543.6,722.1,P＜0.01),并且48 h的IC50为69.4μmol/L,72 h的IC50为32.5μmol/L;七叶皂苷钠作用48 h后Akt、p-Akt和细胞周期蛋白依赖性激酶(CDK)2的表达显著降低,并呈现剂量依赖效应.与未经处理的细胞相比,低浓度(5 μmol/L和10 μmol/L)七叶皂苷钠处理时Akt和p-Akt的表达水平已开始降低(F=613.9,P＜0.05),而从15 μmol/L开始,CDK2表达的降低有统计学意义(F=208.9,208.3,P＜0.05).CDK1、CDK6、细胞周期蛋白D以及细胞周期蛋白E的表达在各处理组间差异无统计学意义(P均＞0.05).结论 七叶皂苷钠可抑制人甲状腺未分化癌HTh74细胞的增殖,可能是通过调节Akt、p-Akt以及CDK2等蛋白的表达实现的.     

Inhibitory effects of extracellular adenosine triphosphate on growth of esophageal carcinoma cells

AIM: To study the growth inhibitory effects of ATP on TE-13 human squamous esophageal carcinoma cells in vitro. METHODS: MTT assay was used to determine the inhibition of proliferation of ATP or adenosine (ADO) on TE-13 cell line. The morphological changes of TE-13 cells induced by ATP or ADO were observed under fluorescence light microscope by acridine orange (A0)/ethidium bromide (EB) double stained cells. The internucleosomal fragmentation of genomic DNA was detected by agarose gel electrophoresis. The apoptotic rate and cell cycle after treatment with ATP or ADO were determined by flow cytometry. RESULTS: ATP and ADO produced inhibitory effects on TE-13 cells at the concentration between 0.01 and 1.0 mmol/L. The IC50 of TE-13 cells exposed to ATP or ADO for 48 and 72 h was 0.71 or 1.05, and 0.21 or 0.19 mmol/L, respectively. The distribution of cell cycle phase and proliferation index (PI) value of TE-13 cells changed, when being exposed to ATP or ADO at the concentrations of 0.01, 0.1, and 1 mmol/L for 48 h. ATP and ADO inhibited the cell proliferation by changing the distribution of cell cycle phase via either G0/G1 phase (ATP or ADO, 1 mmol/L) or S phase (ATP, 0.1 mmol/L) arrest. Under light microscope, the tumor cells exposed to 0.3 mmol/L ATP or ADO displayed morphological changes of apoptosis. A ladder-like pattern of DNA fragmentation was obtained from TE-13 cells treated with 0.1-1 mmol/L ATP or ADO in agarose gel electrophoresis. ATP and ADO induced apoptosis of TE-13 cells in a dose-dependent manner at the concentration between 0.03 and 1 mmol/L. The maximum apoptotic rate of TE-13 cells exposed to ATP or ADO for 48 h was 16.63% or 16.9%, respectively. CONCLUSION: ATP and ADO inhibit cell proliferation, arrest cell cycle, and induce apoptosis of TE-13 cell line.     

5-Fu对低氧下胃癌SGC7901细胞系中SP细胞比例及HIF-2、ABCG2表达的影响

目的:探讨低氧下5-Fu化疗抵抗的机制.方法:MTT检测常氧和低氧下的细胞活力,并计算5-Fu的半数抑制浓度(IC50).以IC50的5-Fu分别作用细胞常氧和低氧组细胞24、48和72h后,收集标本,Hoechst33342染色法检测侧群(side population,SP)细胞的比例,免疫细胞化学法检测低氧诱导因子(hypoxia-inducible factor-2α,HIF-2α),荧光免疫细胞化学法检测ABCG2(ATP-binding cassettesuperfamily Gmember 2)的表达.结果:常氧和低氧下,5-Fu均呈时间、剂量依赖性地抑制SGC7901细胞增殖,其IC50分别为100、200mg/L.常氧下SP细胞的比例为1.87%,低氧诱导后其比例逐渐增加.常氧下5-Fu-IC50作用于细胞不用时间后,SP细胞的比例无明显变化,低氧下其比例却逐渐增加.常氧下,HIF-2和ABCG2蛋白呈低水平表达,且5-Fu-IC50作用于不同时间后也无明显变化,低氧下5-Fu-IC50作用于不同时间后二者的表达逐渐增加.结论:低氧下5-Fu对胃癌SGC7901细胞存在化疗抵抗可能与低氧通过诱导HIF-2α-ABCG2通路的表达、促进肿瘤细胞的干细胞化有关,这可能是肿瘤化疗抵抗和复发的根源.     

Effects of ursolic acid and oleanolic acid on human colon carcinoma cell line HCT15

AIM：Ursolic acid(UA) and oleanolic acid(OA) are triperpene acids having a similar chemical structure and are distributed wildly in plants all lover the world.Iecent years,it was found that they had marked anti-tumor effects,There is little literature currently available regrading their effects on colon carcinoma cells.The present study was designed to investigate their inhibitory effects on human colon carcinoma cell line HCT 15. METHODS：HCT15 cells were cultured with different drugs,The treated cells were stained with hematoxylin-eosin and their morphologic changes observed under a light microscope,The cytotoxicity of these drugs was evaluated by tetrazolium dye assay.Cell dcycle analysis was performed by flow cytomery(FCM),Data were expressed as means±SEM and Analysis of variance and Student‘ t-test for individual comparisons.RESULTS:Twenty-four to 72 h after UA or OA60μmol/L treatment,the numbers of dead cells and cell fragments were increased and most cells were dead at the 72 nd hour.The cytotoxicity of UA was stronger than that of OA.Seventy-eight hours after 30μmol/L of UA or OA treatment a number of cells were degenerated,but cell fragments were rarely seen.The IC50 values for UA and OA were 30 and 60μmol/L,respectively,Proliferation assay showed that proliferation of UA and OA-treated cells was slightly increased at 24h and signficantly decreased at 48h and 60h.whereas untreated control cells maintained an exponential growth curve,Cell cycle analysis by FCM showed HCT15 cells treated with UA 30 and OA 60 for 36h and 72h gradually accumulated in G0/G1 phase(both drugs P<0.05 for 72h),with a concomitant decrease of cell populations in S phase(both drugs P<0.01 for 72h)and no detectable apoptotic fraction.CONCLUSION:UA and OA have significant anti-tumor activity.The effect of UA is stronger than that of OA.The possible mechanism of action is that both drugs have an inhibitory effect on tumor cell proliferation through cell-cycle arrest.     

天然药物IHA-01筛选敏感肿瘤细胞株及其对结肠癌细胞增殖的影响

目的筛选出对天然药物IHA-01的敏感肿瘤细胞,并对其抗瘤机制进行初步研究。方法体外培养12种人肿瘤细胞株,经3.125、6.25、12.5、25和50μg/ml 5个浓度IHA-01作用48 h后光镜下观察细胞株形态变化,采用CCK-8法计算IC50值（半数抑制浓度）,确认敏感细胞株及最佳作用浓度。流式细胞仪检测IHA-01最佳作用浓度下敏感细胞株细胞凋亡情况及端粒酶活性。结果 5个浓度IHA-01均能抑制12种癌细胞增殖,确定结肠癌HCT-8细胞为敏感细胞（IC50值最低）,最佳作用浓度为1.29μg/ml;鼻咽癌CNE-2细胞为不敏感细胞（IC50值最高）。1.29μg/ml的IHA-01作用后HCT-8细胞凋亡率明显高于、端粒酶活性明显低于CNE-2细胞（P均〈0.01）。结论 IHA-01对HCT-8细胞有较强的抑制作用;其机制可能为诱导肿瘤细胞凋亡及抑制细胞端粒酶活性。     

Chemoembolization of rat liver metastasis with irinotecan and quantification of tumor cell reduction

Purpose Hematogenic metastasis of patients with colorectal cancer most frequently effects the liver; the prognosis of affected patients is dramatically worsened by the presence of this lesion. The aim of this study was to evaluate the effect of hepatic arterial chemoembolization (HACE) with irinotecan versus 5-fluorouracil as a standard agent in a rat liver metastasis model.Materials and methods Diffuse liver metastasis was induced by injecting 4×10⁶ CC531-lac-Z rat colorectal carcinoma cells into the portal vein of male Wag/Rij rats. Irinotecan (10 mg/kg, 30 mg/kg, and 60 mg/kg) and 5-fluorouracil (40 mg/kg, 60 mg/kg, and 90 mg/kg) were administered concomitantly with degradable starch microspheres (30 mg/kg) for temporary embolization. The tumor cell load was determined quantitatively using a chemoluminescence assay.Results HACE with irinotecan induced a complete remission in 44% of the animals and the highest dose reduced the mean tumor cell load by 66% (P <0.001). In contrast, the highest dose of 5-FU caused a reduction of only 18% (P = 0.026) and altogether 23% complete remissions were observed in response to 5-FU. The sensitivity of CC531-lac-Z cells versus irinotecan (IC50 32 pM after 72 h) and 5-FU (IC50 80 µM) mirrored the effects observed in vivo on a qualitative basis.Conclusion In conclusion, the effect of HACE with irinotecan surpassed that of HACE with 5-FU and prompts further investigation in clinical trials.     

初步探索DIM-C-pPhOH（ NR4A1拮抗剂）对胶质瘤细胞增殖、迁移及侵袭的抑制作用及作用机制

目的探索DIM-C-pPhOH（NR4A1拮抗剂）对胶质瘤细胞增殖、迁移、侵袭的抑制作用及其作用机制，以及对于胶质瘤小鼠模型生存期及肿瘤生长的影响。 方法不同浓度DIM-C-pPhOH处理胶质瘤细胞系（GL261、U251、U118）48 h后，用CellTiter法检测细胞活性及IC₅₀；采用划痕实验和3D-invasion实验检测DIM-C-pPhOH对U251细胞系侵袭迁移作用的影响；通过腹腔注射该化合物治疗小鼠胶质瘤模型，观察DIM-C-pPhOH对于小鼠胶质瘤生长以及生存期的影响。 结果U251、GL261和U118胶质瘤细胞系经DIM-C-pPhOH处理48 h后，细胞活性随药物浓度增加显著降低，IC₅₀分别为5.76、6.87、9.93 μmol/L；U251细胞系经10 μmol/L DIM-C-pPhOH处理后其迁移和侵袭能力显著下降；同时DIM-C-pPhOH可以抑制由TGF-β诱导的NR4A1出核表达；小鼠胶质瘤模型中DIM-C-pPhOH治疗组生存期延长，肿瘤生长受到抑制。蛋白免疫印迹显示DIM-C-pPhOH可以明显抑制Akt、P70S6K蛋白表达，通过抑制PI3K/Akt/mTOR/p70s6k信号通路，诱导细胞自噬发生。 结论DIM-C-pPhOH可以抑制胶质瘤的迁移及侵袭能力，延长小鼠胶质瘤模型的生存期并抑制肿瘤生长，作用机制可能与DIM-C-pPhOH诱导细胞发生自噬有关。     

帕瑞昔布对人胰腺癌细胞株BxPC-3、AsPC-1增殖凋亡的影响及其机制

目的:研究帕瑞昔布(parecoxib,PCB)对人胰腺癌细胞株BxPC-3和AsPC-1的增殖、凋亡及其可能的分子机制.方法:BxPC-3、AsPC-1细胞用不同浓度PCB的培养液孵育后,利用MTT法测定细胞活性,计算IC50值,TUNEL法检测处理后细胞的凋亡情况,RT-PCR验证相关蛋白的变化表达.结果:PCB对两种细胞生长呈时间和计量依赖性抑制;PCB处理后BxPC-3、AsPC-1两种细胞IC50值为:400.98μmol/L±10.78μmol/L、256.3μmol/L±2.98μmol/L;TUNEL法检测证明凋亡率增加;RT-PCR显示COX-2表达明显降低.结论:PCB可以抑制胰腺癌细胞增殖,并诱导其凋亡生长,其可能机制是通过抑制COX-2表达来实现的.     

Antiproliferation and apoptosis induction of paeonol in HepG2 cells

AIM To investigate the antiproliferative effect of paeonol (Pae) used alone or in combination with chemotherapeutic agents [cisplatin (CDDP), doxorubicin (DOX) and 5-fluorouracil (5-FU)] on human hepatoma cell line HepG2 and the possible mechanisms.METHODS The cytotoxic effect of drugs on HepG2 cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetra-zolium bromide (MTT) assay.Morphologic changes were observed by acridine orange (AO) fluorescence staining. Cell cycle and apoptosis rate were detected by flow cytometry (FCM). Drug-drug interactions were analyzed by the coefficient of drug interaction (CDI).RESULTS Pae (7.81-250 mg/L) had an inhibitory effect on the proliferation of HepG2 cells in a dose-dependent manner, with the IC50 value of (104.77 7.28) mg/L. AO fluorescence staining and FCM assays showed that Pae induced apoptosis and arrested cell cycle at S phase in HepG2 cells. Further, different extent synergisms were observed when Pae (15.63, 31.25, 62.5 mg/L) was combined with CDDP (0.31-2.5 mg/L), DOX (0.16-1.25 mg/L), or 5-FU (12.5-100 mg/L) at appropriate concentrations. The IC50 value of the three drugs decreased dramatically when combined with Pae (P ＜0.01). Of the three different combinations, the sensitivity of cells to drugs was considerably different.CONCLUSION Pae had a significant growth-inhibitory effect on the human hepatoma cell line HepG2,which may be related to apoptosis induction and cell cycle arrest. It also can enhance the cytotoxicity of chemotherapeutic agents on HepG2 cells, and the S phase arrest induced by Pae may be one of the mechanisms of these interactions.     

Anti-hepatocarcinoma effects of 5-fluorouracil encapsulated by galactosylceramide liposormes in vivo and in vitro

AIM: To study the anti-hepatocarcinoma effects of 5fluorouracil (5-Fu) encapsulated by galactosylceramide liposomes (5-Fu-GCL)in vivo and in vitro. METHODS: Tumor-bearing animal model and HepA cell line were respectively adopted to evaluate the anti-tumor effects of 5-Fu-GCL in vivo and in vitro. Tumor cell growth inhibition effects of 5-Fu-GCL in vitro were assessed bycell viability assay and MTT assay. In vivo experiment, the inhibitory effects on tumor growth were evaluated by tumor inhibition rate and animal survival days. High performance liquid chromatography was used to detect the concentration-time course of 5-Fu-GCL in intracellular fluidin vitro and the distribution of 5-Fu-GCL in liver tumor tissues in vivo. Apoptosis and cell cycle of tumor cells were demonstrated by flow cytometry.RESULTS: In vitro experiment, 5-Fu-GCL (6.25-100 μmol/L) and free 5-Fu significantly inhibited HepA cell growth. Furthermore, IC50 of 5-Fu-GCL (34.5 μmol/L) was lower than that of free 5-Fu (51.2 μrnol/L). In vivo experiment, 5-Fu-GCL (20, 40, 80 mg/kg) significantly suppressed the tumor growth in HepA bearing mice model. Compared with free 5-Fu, the area under curve of 5-Fu-GCL in intracellular fluid increased 2.6 times. Similarly, the distribution of 5-Fu-GCL in liver tumor tissues was significantly higher than that of free 5-Fu. After being treated with 5-Fu-GCL, the apoptotic rate and the proportion of HepA cells in the S phase increased, while the proportion in the G0/G1 and G2/M phases decreased. CONCLUSION: 5-Fu-GCL appears to have anti-hepatocarcinoma effects and its drug action is better than free 5-Fu. Its mechanism is partly related to increased drug concentrations in intracellular fluid and liver tumor tissues, enhanced tumor cell apoptotic rate and arrest of cell cycle in S phase.