索拉菲尼治疗肝癌常见不良反应及处理的研究进展

索拉菲尼是一种口服多激酶抑制剂。通过作用于Raf激酶直接抑制肿瘤细胞增殖,还可作用于血管内皮生长因子受体1,2,3（VEGFR-1, -2, -3）,以及血小板源生长因子受体-β（PDGFR-β）、受体酪氨酸激酶、抑制肿瘤新生血管生成。索拉菲尼通过抑制肿瘤细胞增殖和抗血管生成的双重作用,从而达到抗肿瘤的目的。已被多个国家批准作为首个系统治疗肝细胞肝癌的分子靶向药物。其常见不良反应包括皮肤反应、恶心、腹泻、体质量减轻、高血压等,影响了患者的长期使用依从性,进而影响治疗效果。正确地认识和管理索拉非尼的不良反应则有助于发挥索拉非尼的治疗作用,提高临床效果。本文从索拉菲尼靶向治疗的常见不良反应、发生机制及处理方法等方面进行综述。      

索拉菲尼治疗晚期肝癌完全缓解1例

患者男,54岁,2010年9月体检。CT检查提示肝右叶病变,病灶最大直径约8cm,门脉右支癌栓形成,AFP>1 210 mg/mL(图1)。根据AASLD(美国肝病研究学会)诊断标准,临床诊断为HCC。入院后行TACE及门脉支架置入术,2010年10月开始口服索拉菲尼(400 mg/次,2次/d)治疗,截至目前患者AFP水平持续下降,影像学评估显示病灶稳定(图2)。     

线粒体自噬在肝癌发生和治疗中的作用及机制

摘 要：线粒体自噬是机体通过特异性清除自身异常线粒体以促进细胞生存的一种保护性机制。目前大量研究证明,线粒体自噬通过维持细胞稳态在肝癌的发生发展和演化过程中发挥重要作用,并且可以通过多种途径影响肝癌的治疗效果,这为研究肝癌的分子机制及探索新的靶点与治疗手段提供了新的思路。本文将对线粒体自噬在肝癌发生发展和治疗中的作用及机制进行综述,以期深入了解线粒体自噬在肝癌中的重大影响和潜在的治疗价值。     

细胞自噬在肿瘤中的作用

自噬为一种细胞的自我消耗过程,其具有潜在的抗肿瘤生物学活性,亦与肿瘤耐药有关.该文从自噬的调节机制入手,综述自噬参与肿瘤发生的机制,自噬在肿瘤治疗中的作用及应用,以期为自噬在临床上的应用提供依据.     

靶向肝癌细胞自噬提高大黄素的毒性杀伤作用

背景与目的:大黄素处理肝癌细胞后能够诱导内质网应激和凋亡.鉴于内质网应激与自噬之间的关联及后者作为细胞对抗应激环境的一种自我防御机制,该研究拟探讨通过抑制肝癌细胞自噬信号通路的策略提高大黄素对肿瘤细胞的毒性杀伤作用.方法:大黄素处理肝癌细胞后,应用CYTO-ID自噬检测试剂盒和蛋白[质]印迹法(Western blot)分别检测大黄素诱发细胞自噬情况;利用细胞自噬抑制剂(氯喹)预先抑制肝癌细胞自噬的产生,然后用大黄素处理肝癌细胞,最后通过ATPlite试验和细胞克隆形成实验检测肿瘤细胞存活;通过流式细胞术检测氯喹和大黄素联合处理诱导肝癌细胞发生凋亡的凋亡率,采用Western blot检测凋亡效应蛋白caspase-3活化断裂后产生活性片段的水平.结果:大黄素处理肝癌细胞后能够诱导肝癌细胞自噬;利用氯喹抑制肝癌细胞自噬能够显著能够提高大黄素对肝癌细胞克隆存活的抑制作用;氯喹和大黄素联合处理肝癌细胞能够显著提高细胞周期sub-G1期和活化caspase-3蛋白的表达水平.结论:靶向肝癌细胞自噬能够提高大黄素的毒性杀伤作用.     

桧木醇激活膀胱癌J82细胞自噬诱导细胞凋亡

背景与目的：膀胱癌是我国最常见的泌尿系统恶性肿瘤,近年来发病率逐年上升,严重威胁着人类的健康。本研究旨在探讨桧木醇对人膀胱癌J82细胞增殖、凋亡及自噬的作用,并初步阐明其作用机制。方法：采用CCK-8法检测细胞的增殖活力,采用流式细胞术检测细胞的凋亡状态,采用蛋白[质]印迹法(Western blot)检测cleaved caspase 3、LC3及P62蛋白的表达,转染EGFP-LC3后在激光共聚焦显微镜下观察细胞自噬状态。结果：桧木醇能够显著抑制J82细胞的增殖活力,通过激活caspase途径诱导肿瘤细胞凋亡,Z-VAD-FMK能够部分抑制桧木醇的凋亡诱导作用。桧木醇能够激活J82细胞发生自噬,上调LC3蛋白的表达,下调P62蛋白的表达。3-MA抑制自噬之后能够部分逆转桧木醇的抗肿瘤作用。结论：桧木醇可以显著抑制J82细胞增殖并通过过度激活自噬促进膀胱癌细胞凋亡。     

甲胎蛋白与自噬在肝癌发生发展中的作用及其相互关系

甲胎蛋白（AFP）作为临床诊断肝癌最常用的肿瘤标志物,有抑制免疫、促进细胞生长、抑制癌细胞凋亡的作用。自噬是一种维持细胞生存的重要途径之一,其与肝癌的发生发展及治疗有着密切联系,对肝癌既有抑制又有促进作用。PI3K/AKT作为两者共有的信号通路,它们是否有着相互关系来促进肝癌的发展尚需进一步研究。     

晚期肝癌索拉菲尼治疗完全缓解1例报告简

晚期肝癌缺乏有效的治疗方法.近几年来,索拉菲尼用于晚期肝癌的临床治疗,两个大型临床试验证实对晚期肝癌患者具有明确的临床疗效[1,2],但有关索拉菲尼治疗晚期肝癌获得完全缓解（CR）的报道甚少.作者应用索拉菲尼治疗晚期肝癌,1例获得完全缓解,无瘤生存时间超过2年,现报道如下.     

自噬在肝癌介入治疗中的研究进展

原发性肝癌是我国最常见的恶性肿瘤之一,介入治疗覆盖中国肝癌分期Ⅰa～Ⅲb期的患者。自噬作为真核细胞特有的代谢过程,参与代谢性疾病、恶性肿瘤的发生、发展,在肝癌的发生、发展中发挥重要作用,尤其促进了肿瘤细胞对介入治疗的耐受,使得治疗效果不佳。本文综述了原发性肝癌常见介入治疗方式对肿瘤细胞自噬现象的影响：消融治疗、经血管介入治疗及内放射治疗都会通过缺氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)/B淋巴细胞瘤2/腺病毒E1B相互作用蛋白3(B-cell lymphoma-2/adenovirus E1B 19-kDa-interacting protein 3,BNIP3)通路、磷脂酰肌醇3激酶（phosphoinositide 3-kinase,PI3K）/蛋白激酶B(protein kinase B,AKT)/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin,mTOR）等细胞通路诱导肿瘤细胞自噬水平的提高,从而产生治疗抵抗。体外试验中,自噬抑制剂的使用可以改善自噬引起的治疗抵抗。     

索拉非尼治疗晚期原发性肝癌的临床观察

目的　观察索拉非尼治疗晚期原发性肝癌的疗效、生活质量以及不良反应。方法　２０例无法手术的原发性肝癌患者口服索拉非尼（４００ｍｇｂｉｄ）,至出现不可耐受的不良反应或连续两次评价为疾病进展,观察客观疗效、无进展生存期（ＴＴＰ）、总生存期（OＳ）、生活质量和不良反应。结果　２０例患者中ＳＤ１５例,ＰＤ５例,中位ＴＴＰ为５.９月（１.５～１５.０个月）,中位ＯＳ为７.７个月（２.０～２１.０个月）。服药期间患者的总体健康状况及五项功能领域得分在第６周时下降,但自第１２周上述功能领域均能稳定在治疗前状态。疼痛及腹泻在治疗过程中持续存在,而乏力、恶心呕吐、失眠、厌食、便秘及经济困难均得以改善。最常见的不良反应是手足皮肤反应,食欲下降和腹泻。结论　索拉非尼用于晚期原发性肝癌患者疗效肯定,不良反应少且多数可耐受,服药期间患者的生活质量能保持稳定。     

Proteasome inhibitor interacts synergistically with autophagy inhibitor to suppress proliferation and induce apoptosis in hepatocellular carcinoma

BACKGROUND:

The ubiquitin‐proteasome system and autophagy‐lysosome system are 2 major protein degradation pathways in eukaryotic cells, which are tightly linked to cancer. Proteasome inhibitors have been approved in clinical use against hematologic malignancies, but their application in solid tumors is uncertain. Moreover, the role of autophagy after proteasome inhibition is controversial.

METHODS:

Two proteasome inhibitors, 2 autophagy inhibitors, and 3 hepatocellular carcinoma (HCC) cell lines were investigated in the current study. In vitro, cell proliferation was evaluated by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay, cell apoptosis was evaluated by flow cytometry analysis of annexin‐V/propidium iodide staining, and autophagy was evaluated by green fluorescent protein‐light chain 3 (GFP‐LC3) redistribution and LC3 Western blot analysis. In vivo, Ki‐67 staining was used to detect cell proliferation, terminal deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate nick‐end labeling (TUNEL) staining was used to detect apoptosis, and electron microscopy and p62 immunohistochemical staining were used to detect autophagy.

RESULTS:

Proteasome inhibitors suppressed proliferation, induced apoptosis, and activated autophagy in HCC cell lines in vitro, and autophagy exerted a protective role after proteasome inhibition. In vivo, anticancer effects of bortezomib on the MHCC‐97H orthotopic model (human HCC cells) were different from the effects observed on the Huh‐7 subcutaneous model (human HCC cells). The autophagy inhibitor chloroquine interacted synergistically with bortezomib to suppress proliferation and induce apoptosis in both tumor models.

CONCLUSIONS:

The current results indicated that simultaneous targeting of the proteasome and autophagy pathways may represent a promising method for HCC treatment. Cancer 2012. © 2012 American Cancer Society.     

SNCG在肝癌高/低淋巴道转移细胞株中的差异性表达

目的:研究SNCG蛋白在小鼠肝癌高/低淋巴道转移潜能细胞株中的定位和表达差异。方法:应用免疫细胞化学、Western blot方法检测SNCG蛋白在小鼠肝癌高/低淋巴道转移潜能细胞株Hca-F/Hca-P的定位情况和表达差异。结果:SNCG蛋白在Hca-F和Hca-P细胞株中均主要定位于细胞浆,细胞核区域也可见少量蛋白表达。SNCG在Hca-F中的表达［染色强度为(+++)］强于在Hca-P中的表达［染色强度为(++)］。检测到一条SNCG特异性蛋白条带,在Hca-F细胞中表达量为Hca-P细胞中的1.9倍。结论:SNCG的高表达可能与高淋巴道转移潜能有关。     

Sorafenib for Egyptian patients with advanced hepatocellular carcinoma; single center experience

BackgroundAccording to the results of a number of phase 3 randomized studies, sorafenib is the only approved systemic therapy for advanced HCC; however the issue of high economic cost remains challenging; thus we have conducted this retrospective analysis of our HCC patients treated with sorafenib.MethodsHCC patients treated at Ain Shams University Hospitals, in the period between 2010 and 2012 were reviewed. Eligible patients were those who had received sorafenib for advanced HCC not eligible for or progressed after surgery or locoregional therapy. We investigated the impact of baseline clinicopathological factors (age, gender, child status, performance score, BCLC tumor stage, cause of chronic liver disease, median baseline alpha fetoprotein level and previous treatment received for HCC) on overall survival (OS) in an adjusted Cox regression model.Results41 patients were included in the analysis fulfilling the inclusion criteria. At a median follow up period of 13 months, the median PFS for the whole group was 4 months; the median OS for the whole group is 6.25 months. Multivariate analysis identified three baseline characteristics that were prognostic indicators for overall survival: ECOG performance status (median OS for ECOG 1 = 7.01 months and for ECOG 2 = 3.03 months), Child–Pugh status (median OS for child A = 12.04 months and for child B = 5.23 months), and median baseline levels of alpha-fetoprotein.ConclusionsIn limited resource countries like Egypt, we suggest that the use of sorafenib for the treatment of advanced HCC cases should be restricted to a highly selected subgroup of patients with good performance and child A.     

Sorafenib combined with cryoablation to treat unresectable hepatocellular carcinoma

Objective  

To evaluate the efficacy and tolerability of sorafenib combined with cryoablation in treating unresectable hepatocellular carcinoma (HCC).     

Nitric oxide inhibits autophagy and promotes apoptosis in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the second most common cause of cancer‐related mortality worldwide. The expression of nitric oxide synthase (NOS) and the inhibition of autophagy have been linked to cancer cell death. However, the involvement of serum nitric oxide (NO), the expression of NOS and autophagy have not been investigated in HCC. In the present study, we first established that the NO level was significantly higher in hepatitis B virus‐related HCC than in the liver cirrhosis control (53.60 ± 19.74 vs 8.09 ± 4.17 μmol/L, t = 15.13, P < 0.0001). Using immunohistochemistry, we found that the source of NO was at least partially attributed to the expression of inducible NOS and endothelial NOS but not neuronal NOS in the liver tissue. Furthermore, in human liver cancer cells, NO‐induced apoptosis and inhibited autophagy. Pharmacological inhibition of autophagy also induced apoptosis, whereas the induction of autophagy could ameliorate NO‐induced apoptosis. We also found that NO regulates the switch between apoptosis and autophagy by disrupting the Beclin 1/Vps34 association and by increasing the Bcl‐2/Beclin 1 interaction. Overall, the present findings suggest that increased NOS/NO promotes apoptosis through the inhibition of autophagy in liver cancer cells, which may provide a novel strategy for the treatment of HCC.     

经导管升压化疗栓塞后二期肝切除肝癌细胞凋亡的研究

 本文报告11例经导管血管紧张素Ⅱ介导升压化疗栓塞（IHCE）后二期切除肝癌标本中残癌细胞凋亡的研究，并与9伊TCE后二期切除标本及10例一期肝癌切除标本比较。结果：在H·E染色标本中，IHCE,TCE组较易见到凋亡细胞，而一期切除肝癌细胞中极少见。用流式细胞仪进行定量分析，IHCE的残癌细胞调亡为42.8±27.4%,TCE组25.3±10.8%,一期切除组4.9±3.4%.     

siRNA沉默RhoC表达诱导人肝癌BEL7402细胞凋亡

目的:研究siRNA沉默RhoC基因表达对人肝癌细胞BEL7402凋亡的影响及其机制,为肝癌的基因治疗提供实验依据。方法:构建RhoC-siRNA真核表达载体pU6mRFPRhoC-siRNA,转染BEL7402细胞,激光共聚焦显微镜检测转染效率,RT-PCR和Western blotting鉴定RhoC基因沉默效果;流式细胞术、琼脂糖凝胶电泳和瑞氏染色检测BEL7402细胞凋亡,RT-PCR检测细胞凋亡相关基因Bcl-2和Bax的表达。结果:成功构建pU6mRFPRhoC-siRNA重组载体,转染BEL7402细胞的效率为70%,RT-PCR和Western blotting检测RhoC基因沉默效率分别为85%和82%。pU6mRFPRhoC-siRNA转染组BEL7402细胞凋亡显著高于未转染BEL7402细胞和pU6mRFP Scramble-siRNA转染组BEL7402细胞[(21.00±2.23)%vs(6.47±1.64)%、(4.63±0.47)%,P<0.01)],pU6mRFPRhoC-siRNA转染组BEL7402细胞DNA呈典型的梯状条带,瑞氏染色见转染组BEL7402细胞中有大量凋亡...     

TRAIL受体在人肝细胞癌中的表达及TRAIL诱导的肝癌细胞凋亡

目的 研究了RAIL受体在肝癌组织及肝癌细胞中的表达,探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体在肝癌细胞凋亡诱导中的作用。 方法 应用RT-PCR法和免疫组化方法检测肝癌组织及细胞系SK-Hepl中TRAIL受体的表达,并以TRAIL蛋白和γ-干扰素作用于SK-Hepl细胞,观察其对细胞的凋亡诱导作用。 结果 在肝癌组织及肝癌SK-Hepl细胞系中均可检测到TRAIL受体,其中,死亡受体DR4、DR5在肝癌、癌旁组织以及SK-Hepl肝癌细胞系中均呈高表达,而诱骗受体DcR1及DcR2呈低表达,明显低于正常组织。TRAIL对肝癌细胞有凋亡诱导作用,与干扰素的协同时,其作用明显增强。 结论 肝癌组织中存在着TRAIL受体的表达,TRAIL可诱导肝癌SK-Hepl细胞系凋亡,其作用与DR4、DR5的高表达有关。