p16与c-myc基因表达产物在肝细胞癌及癌旁组织中的表达及临床意义

目的：探讨ｐ１６、ｃ－ｍｙｃ蛋白与肝细胞癌（ＨＣＣ）的发生、发展及恶性程度的关系。方法：应用免疫组织化学法检测了６９例ＨＣＣ和６３例癌旁组织ｐ１６及ｃ－ｍｙｃ蛋白的表达,并结合临床资料进行分析。结果：ｐ１６蛋白主要分布在胞浆,少数细胞核内也可见阳性显色。ｃ－ｍｙｃ蛋白主要分布在胞核。ｐ１６蛋白在肝癌组织及癌旁组织中的阳性率分别为２６．５％（１８／６９）和９２％（５８／６３）,两者比较差异显著（Ｐ〉     

Thymidine transport in hepatocytes

Thymidine transport and phosphorylation were investigated in isolated rat hepatocytes and AS 30 D hepatoma cells. In contrast to hepatoma cells, hepatocytes exhibited a minimum of thymidine phosphorylation due to a 100-fold smaller thymidine kinase activity. In hepatocytes thymidine is transported by two transport systems: a specific concentrative high affinity system and an unspecific non-concentrative low affinity system. In hepatoma cells only the low affinity system could be detected. A single dose of 20 or 50 mg diethylnitrosamine/kg body weight induced in hepatocytes a remarkable increase of thymidine kinase activity and a decrease of the transport by the high affinity system. Thymidine transport and phosphorylation by hepatocytes are considered to be sensitive markers for early recognition of toxin-induced liver regeneration.

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Zusammenfassung Thymidintransport und -phosphorylierung wurden in isolierten Rattenhepatozyten und AS 30 D-Hepatomzellen untersucht. Hepatozyten wiesen im Gegensatz zu Hepatomzellen aufgrund einer 100fach niedrigeren Thymidinkinaseaktivität eine äußerst niedrige Thymidinphosphorylierungsrate auf. In Hepatozyten wurde Thymidin durch zwei Transportsysteme aufgenommen: ein spezifisches, konzentratives high affinity System und ein unspezifisches, nichtkonzentratives low affinity System. In ATP-freien Hepatomzellen konnte nur das low affinity System nachgewiesen werden. Eine einmalige Dosis von 20 bzw. 50 mg Diäthylnitrosamin/kg Körpergewicht bewirkte in Hepatozyten einen Anstieg der Thymidinkinaseaktivität und eine Verminderung des Thymidintransports über das high affinity System. Thymidintransport und -phosphorylierung in Hepatozyten erwiesen sich als sensitives System zur frühen Erkennung beginnender Leberregeneration nach toxischer Vorschädigung.

     

全反式维甲酸抑制Ets-1介导的肝癌7402细胞离散和侵袭及其分子机制

目的 研究全反式维甲酸(ATRA)对Ets介导的实体瘤细胞侵袭转移性的影响及探讨其分子机制,寻找新的抗癌药物作用靶点。方法 采用基因转染的方法,将Ets-1基因高表达在低侵袭转移的肝癌7402细胞中,使其增殖、离散侵袭能力增强,再经ATRA处理细胞,用细胞生物学方法检测细胞增殖性、肝癌细胞离散性及穿透Transwell聚碳酯膜能力的改变。并用Northern杂交、Western印迹等分子生物学手段检测ATRA的分子作用机制。结果 发现ATRA可抑制7402细胞因Ets-1高表达而增强的增殖、离散、侵袭性,可下调Ets、c-Met基因的表达,使MAPK、PI3K信号途径中两个关键分子 Erk/Akt的磷酸化水平降低。结论ATRA可能通过下调Ets基因的表达,进而使c-Met表达水平降低,导致MAPK、PI3K信号转导减弱而抑制肿瘤细胞增殖和侵袭;Ets基因可以作为抗癌药物作用的靶点来筛选药物或进行药理研究。     

人参皂甙Rg3在小鼠肝癌淋巴结转移模型中诱导细胞凋亡的作用

目的:观察人参皂甙Rg3对小鼠肝癌淋巴结转移模型中肿瘤细胞凋亡的诱导作用,探讨人参皂甙Rg3抗肿瘤淋巴结转移的机制.方法:建立小鼠肝癌淋巴结转移模型;光镜和透射电镜下观察各组(Rg3预防组、Rg3治疗组、Rg3与顺铂联合治疗组、顺铂治疗组及对照组)中原发瘤及转移瘤组织的形态学结构改变,并通过流式细胞仪分析肿瘤细胞的凋亡.结果:Rg3预防组及治疗组电镜下(5份样品)可见较多细胞凋亡小体的形成(5/5,4/5);顺铂治疗组的形态改变以细胞破坏为主,凋亡的细胞较少;联合治疗组细胞的凋亡和坏死程度相当.流式细胞学检测分析结果为:Rg3预防组、治疗组及联合治疗组均见细胞凋亡的特征峰(5/5),而顺铂治疗组为1/5,对照组未见凋亡峰(0/5).结论:人参皂甙Rg3抗肿瘤细胞淋巴结转移的作用与诱导细胞凋亡有关.     

TGF-β1对人肝癌细胞系凋亡的调控作用研究

 目的　本研究旨在明确人肝癌细胞系中TGF β1的作用及其与凋亡的关系。 方法　本研究选用了 3种含有不同 p5 3基因状态的人肝癌细胞系 ,应用TUNEL技术对TGF β1诱导的肝癌细胞的凋亡进行了定量检测。结果　TGF β1仅能诱导HepG2细胞 (野生型p5 3)凋亡 ,这提示凋亡与 p5 3基因的表达具有明确的联系。结论　HepG2细胞系比Huh 7和Hep3B系细胞更易发生TGF β1诱导的凋亡 ,TGF β1通过p5 3依赖性途径诱导肝癌细胞系发生凋亡     

不同p53状态的肝癌细胞系DNA损伤修复能力的比较

目的 观察不同p53状态的肝癌细胞在DNA损伤因素存在下，探讨p53在肝细胞癌发生过程中的作用。方法 选用内源性表达野生型p53、突变型p53、p53全基因缺失的HepG2、PLC／PRF／5、Hep3B肝癌细胞系，将含有p53结合序列的CAT报告基因质粒分别转染各细胞，通过ELISA方法检测报告基因CAT活化情况，观察p53功能状态；用细胞生长曲线比较不同p53状态的细胞生长速度的差异；给予一定剂量的紫外线照射，检测各细胞程序外DNA损伤修复(UDS)能力；观察紫外线照射后细胞克隆形成情况，反映不同细胞系在紫外线照射后细胞存活率的不同。结果 报告基因CAT在HepG2细胞表达最强，而在PLC／PRF／5、Hep3B肝癌细胞均处于较低水平，说明HepG2细胞具有功能性的p53表达；HepG2细胞生长速度明显慢于其他两种细胞；紫外线照射后，HepG2具有较好的DNA损伤修复能力以及较高的细胞生存率。结论 野生型p53具有抑制肝癌细胞生长、保持细胞良好的DNA损伤修复的功能。     

Endocrine Active Compounds: From Biology to Dose Response Assessment

Histone deacetylase (HDAC) inhibitors target key steps of tumor development: They inhibit proliferation, induce differentiation and/or apoptosis, and exhibit potent antimetastatic and antiangiogenic properties in transformed cells in vitro and in vivo. Preliminary studies in animal models have revealed a relatively high tumor selectivity of HDAC inhibitors, strenghtening their promising potential in cancer chemotherapy. Until now, preclinical in vitro research has almost exclusively been performed in cancer cell lines and oncogene-transformed cells. However, as cell proliferation and apoptosis are essential for normal tissue and organ homeostasis, it is important to investigate how HDAC inhibitors influence the regulation of and interplay between proliferation, differentiation, and apoptosis in primary cells as well. This review highlights the discrepancies in molecular events triggered by trichostatin A, the reference compound of hydroxamic acid-containing HDAC inhibitors, in hepatoma cells and primary hepatocytes (which are key targets for drug-induced toxicity). The implications of these differential outcomes in both cell types are discussed with respect to both toxicology and drug development. In view of the future use of HDAC inhibitors as cytostatic drugs, it is highly recommended to include both tumor cells and their healthy counterparts in preclinical developmental studies. Screening the toxicological properties of compounds early in their development process, using a battery of different cell types, will enable researchers to discard those compounds bearing undesirable adverse activity before entering into expensive clinical trials. This will not only reduce the risk for harmful exposure of patients but also save time and money.     

Antiangiogenic activity of 2-formyl-8-hydroxy-quinolinium chloride

Tumour growth is closely related to the development of new blood vessels to supply oxygen and nutrients to cancer cells. Without the neovascular formation, tumour volumes cannot increase and undergo metastasis. Antiangiogenesis is one of the most promising approaches for antitumour therapy. The exploration of new antiangiogenic agents would be helpful in antitumour therapy. Quinoline is an aromatic nitrogen compound characterized by a double-ring structure which exhibits a benzene ring fused to pyridine at two adjacent carbon atoms. The high stability of quinoline makes it preferable in a variety of therapeutic and pharmaceutical applications, including antitumour treatment. This work is to examine the potential antiangiogenic activity of the synthetic compound 2-Formyl-8-hydroxy-quinolinium chloride. We found that 2-Formyl-8-hydroxy-quinolinium chloride could inhibit the growth of human umbilical vein endothelial cells in vitro. Using the diethylnitrosamine-induced hepatocarcinogenesis model, 2-Formyl-8-hydroxy-quinolinium chloride showed strong antiangiogenic activity. Furthermore, 2-Formyl-8-hydroxy-quinolinium chloride could inhibit the growth of large Hep3B xenografted tumour from the nude mice. We assume that 2-Formyl-8-hydroxy-quinolinium chloride could be a potential antiangiogenic and antitumour agent and it is worthwhile to further study its underlying working mechanism.     

异丙酚持续静脉泵入麻醉对接受微波消融治疗的原发性肝癌患者血管内皮糖萼和脑代谢的影响

目的 探讨应用丙泊酚持续静脉泵注麻醉对接受手术治疗的原发性肝癌(PLC)患者血管内皮糖萼和脑代谢的影响。方法 2017年8月～2020年7月在我院行微波消融术治疗的PLC患者78例,其中一半患者接受异丙酚麻醉,另一半接受地佐辛麻醉。在术前(T0)、术后1 d(T1)和术后7 d(T2),行血气分析,计算脑氧摄取率(CEO₂),采用ELISA法检测硫酸乙酰肝素酶(HS)和多配体蛋白聚糖-1。结果 异丙酚麻醉组诱导时间和苏醒时间分别为(40.3±3.2) s和(6.5±0.8)min,显著短于地佐辛麻醉组【分别为(52.0±3.1) s和(9.8±1.1) min, P<0.05】;在T1和T2,异丙酚麻醉组HS分别为(17.3±2.1)pg/ml和(8.2±0.4)pg/ml,显著低于地佐辛麻醉组【分别为(37.2±5.6)pg/ml和(16.0±2.5)pg/ml,P<0.05】,血清多配体蛋白聚糖-1水平分别为(8.2±0.1)pg/ml和(3.8±0.1)pg/ml, 显著低于地佐辛麻醉组【分别为(11.7±1.4)pg/ml和(5.9±0.2) pg/ml,P<0.05】,CEO₂分别为(28.0±2.4)%和(32.7±3.7)%,显著高于地佐辛麻醉组【分别为(25.0±3.1)%和(29.7±4.1)%,P<0.05】。结论 应用异丙酚持续静脉泵注麻醉在接受微波消融治疗的PLC患者能抑制血管内皮糖萼的激活,改善脑代谢,提高麻醉效果,减少并发症的发生。     

索拉菲尼治疗进展期原发性肝癌患者有效性及安全性研究*

目的 探讨应用索拉菲尼治疗进展期原发性肝癌(aPLC)患者的有效性和安全性。方法 2013年4月~2016年12月我科诊治的aPLC患者82例,采用随机数字表法分为两组,每组41例。两组均给予射频消融术（RFA）治疗,观察组患者在RFA前后接受索拉菲尼治疗,观察12 w。采用ELISA法检测血清碱性成纤维细胞生长因子（bFGF）和血管内皮生长因子（VEGF）。结果 2例观察组患者被剔除,3例对照组患者失访;在治疗12 w末,观察组疾病控制率为61.5%,与对照组的52.6%比,差异无统计学意义（P>0.05）,但观察组肿瘤客观有效率为48.7%,显著高于对照组的26.3%,差异有统计学意义（P<0.05）;观察组血清AFP、bFGF和VEGF水平分别为（184.7±10.5）μg/L、（3.8±1.3） pg/mL和（172.3±25.4） pg/mL,均显著低于对照组的（213.6±11.6） μg/L、（6.4±2.0） pg/mL和（210.5±28.3） pg/mL,差异有统计学意义（P<0.05）;观察组中位无进展生存期（PFS）为10.2个月（95%CI为7.4~11.5）,对照组为7.9个月（95%CI为 6.0~10.1）,经Log-rank检验显示两组差异有统计学意义（P<0.05）;观察组在服药过程中手足综合征、皮疹、白细胞减少、口腔黏膜炎、脱发和肝功能异常发生率分别为43.6%、25.6%、17.9%、20.5%、25.6%和23.1%,显著高于对照组的0.0%、2.6%、0.0%、0.0%、0.0%和5.3%,差异有统计学意义（P<0.05）。结论 索拉菲尼可控制aPLC患者实体瘤扩散,延长无进展生存期,但索拉菲尼可引起多种不良反应,在用药过程中应注意观察并及时采取相应处理措施,以防止发生严重不良反应。