Idarubicin vs doxorubicin in transarterial chemoembolization of intermediate stage hepatocellular carcinoma

BACKGROUND Liver cancer is the fifth most common cancer and the second cause of cancerrelated deaths worldwide.Transarterial chemoembolization(TACE)is the best treatment of intermediate hepatocellular carcinoma(HCC).Doxorubicin is the most commonly used drug despite a low level of evidence.AIM To compare the objective response rate of idarubicin-based TACE(Ida-TACE)against doxorubicin-based TACE(Dox-TACE)in intermediate stage HCC.METHODS Between January 2012 and December 2014,all patients treated with TACE at our academic hospital were screened.Inclusion criteria were patients with Child-Pugh score A or B,a performance status below or equal to 1,and no prior TACE.Either lipiodol TACE or drug-eluting beads TACE could be performed with 10 mg of idarubicin or 50 mg of doxorubicin.Each patient treated with idarubicin was matched with two doxorubicin-treated patients.The TACE response was assessed by independent radiologists according to the mRECIST criteria.RESULTS Sixty patients were treated with doxorubicin and thirty with idarubicin.There were 93%and 87%of cirrhotic patients and 87%and 70%of Child-Pugh A in the doxorubicin and idarubicin groups,respectively.The median number of HCC per patient was two in both groups with 31%and 26%of single nodules in doxorubicin and idarubicin groups,respectively.Objective response rate after first TACE was 76.7%and 73.3%(P=0.797)with 41.7%and 40.0%complete response in doxorubicin and idarubicin groups,respectively.Progression-free survival was 7.7 mo in both groups,and liver transplant-free survival was 24.9 mo and 21.9 mo in doxorubicin and idarubicin groups,respectively.Safety profiles were similar in both groups,with grade 3-4 adverse events in 35%of Dox-TACE and 43%of Ida-TACEs.CONCLUSION Ida-TACE and Dox-TACE showed comparable results in terms of efficacy and safety.Ida-TACE may represent an interesting alternative to Dox-TACE in the management of patients with intermediate stage HCC.     

三苯氧胺在肝癌化疗中作用机制的实验研究

目的 研究三苯氧胺在肝癌化疗中的作用效果，并对其作用机制进行初步探讨。方法 通过阿霉素（ADM）浓度梯度递增诱导法，建立人肝癌多药耐药细胞株Hep-3B／ADM。MTT法检测细胞对化学疗法药物的敏感性；流式细胞仪检测细胞表面多药耐药基因（MDRI）表达产物P-170及分析细胞内Rhdaming123（Rh123）相对荧光强度；流式细胞仪及电镜观察TAM对ADM诱导Hep-3B／ADM细胞凋亡的影响。结果 TAM预处理组ADM对Hep-3B／ADM细胞的IC50下降为非预处理组的1／7；TAM（2．5μmol／L）处理前后，Hep-3B／ADM细胞表面P-170表达及细胞内Rh123相对荧光强度均无明显变化；TAM（2．5μmoL／L）可明显增强ADM诱导Hep-3B／ADM细胞凋亡的效果。结论 TAM（2．5μmol／L）具有增强ADM对Hep-3B／ADM细胞的毒性作用，其作用机制与逆转MDR无关，而是增强了ADM诱导耐药细胞凋亡的作用。     

组蛋白去乙酰化酶1、2在大肠癌及腺瘤组织中的表达及其与临床病理的关系

目的:研究组蛋白去乙酰化酶1(HDAC1)和组蛋白去乙酰化酶2(HDAC2)在大肠正常组织、腺瘤、腺癌组织中的表达,探讨其可能的临床病理及预后的关系.方法:以免疫组织化学SP法检测HDAC1和HDAC2在正常大肠组织、腺瘤、腺癌组织中的表达.通过计算染色指数(SI),评估两者的表达水平与年龄、肿瘤大小、分期、分化等临床病理特征之间的关系.采用Kaplan-Meier法进行生存分析.结果:HDAC1和HDAC2在正常组织表达明显低于腺瘤及癌组织(14.3±9.3vs22.4±12.4,22.8±8.5;5.6±3.3,12.3±4.2vs16.2±9.7,均P<0.05).HDAC2在三者中的表达呈递增趋势.直径≥5cm的癌组织HDAC1的表达高于<5cm的癌组织(25.1±8.2vs20.4±8.5),差异均有统计学意义.结论:HDAC1和HDAC2是大肠癌发生的早期事件,两者共同促进大肠癌的发生发展,可能成为大肠癌治疗的新靶点.     

Effects and mechanisms of silibinin on human hepatocellular carcinoma xenografts in nude mice

AIM： To investigate the in vivo effects and mechanisms of silibinin on the growth of hepatocellular carcinoma （HCC） xenografts in nude mice. METHODS： Nude mice bearing HuH7 xenografts were used to assess the anti-HCC effects and mechanisms of silibinin. RESULTS： Silibinin resulted in a potent dosedependent reduction of HuH7 xenografts in association with a significant decrease in Ki-67 and α-fetoprotein production, nuclear NF-κB content, polo-like kinase 1, Rb phosphorylation, and E2F1/DP1 complex, but increased p27/CDK4 complex and checkpoint kinase 1 expression, suggesting that the in vivo effects of silibinin are mediated by inhibiting G1-S transition of the cell cycle. Silibinin-induced apoptosis of HuH7 xenografts was associated with inhibited survivin phosphorylation. Silibinin-reduced growth of HuH7 xenografts was associated with decreased p-ERK, increased PTEN expression and the activity of silibinin was correlated with decreased p-Akt production, indicating involvement of PTEN/PI3K/Akt and ERK pathways in its in vivo anti-HCC effects. Silibinin-reduced growth of HuH7 xenografts was also associated with a significant increase in AC-H3 and AC-H4 expression and the production of superoxide dismutase （SOD）-1.CONCLUSION： Silibinin reduces HCC xenograft growth through the inhibition of cell proliferation, cell cycle progression and PTEN/P-Akt and ERK signaling, inducing cell apoptosis, and increasing histone acetylation and SOD-1 expression.     

肝细胞癌患者血浆SPINK1水平及其与患者预后的关系分析

目的 分析肝细胞癌（HCC）患者血浆丝氨酸蛋白酶抑制剂Kazal 1型（SPINK1）水平及与其患者术后预后的关系。方法 2014年5月~2015年9月我院收治的HCC患者80例、肝硬化患者40例,选择正常人30例,所有纳入对象在晨起空腹取静脉血,采用ELISA法测定血浆SPINK1和甲胎蛋白（AFP）水平,绘制受试者工作特征曲线（ROC）,评价血浆SPINK1和AFP对HCC的诊断效能,并分析术前血浆SPINK1水平与HCC患者预后的关系。结果 HCC患者血浆SPINK1为（2.4±0.3） μg/L,显著高于肝硬化患者的（2.0±0.2） μg/L或正常人的（1.3±0.1） μg/L,肝硬化患者血浆SPINK1水平显著高于正常人（P<0.05）;HCC患者血浆AFP水平为（125.7±14.5） μg/L,显著高于肝硬化患者的（35.4±4.2）μg/L或正常人的【（3.6±0.5）μg/L,P<0.05】;ROC曲线分析结果显示,以1.70 μg/L为截断点,血浆SPINK1水平诊断HCC的灵敏度为77.2%,特异度为67.6%,而以最佳截断点为124.8μg/L,血浆AFP 水平则分别为63.4%和62.5%;42例术前血浆SPINK1水平>1.7 μg/L的HCC患者术后总生存期（OS）为（12.2±1.3） m,无瘤生存期（DFS）为（11.2±1.2） m,显著短于38例血浆SPINK1<1.7 μg/L组的（20.5±2.1） m和（15.7±1.9） m,而复发率为38.1%,显著高于术前血浆SPINK1<1.7 μg/L患者的18.4%（x²=5.18,P<0.05）。结论 HCC患者血浆SPINK1呈高水平状态,SPINK1诊断HCC的灵敏度、特异度较AFP高,且SPINK1与HCC患者预后密切相关,临床可将其作为HCC肿瘤标志物。     

靶向Plk1的siRNA对肝癌细胞凋亡的影响

目的:观察靶向作用于Plk1的siRNA对肝癌细胞系BCL-7402细胞中Plk1基因和p53基因表达及细胞凋亡的影响,探求靶向该基因的治疗在肝癌基因治疗中的可行性及效应.方法:设计合成两对靶向作用于Plk1基因的双链siRNA序列,分别命名为siRNA1和siRNA2,应用脂质体法将其转入BCL-7402细胞中.实验...     

TRAIL-induced apoptosis of hepatocellular carcinoma cells is augmented by targeted therapies

AIM: To analyze the effect of chemotherapeutic drugs and specific kinase inhibitors, in combination with the death receptor ligand tumor necrosis factor-related apoptosis inducing ligand (TRAIL), on overcoming TRAIL resistance in hepatocellular carcinoma (HCC) and to study the efficacy of agonistic TRAIL antibodies, as well as the commitment of antiapoptotic BCL-2 proteins, in TRAIL-induced apoptosis.METHODS: Surface expression of TRAIL receptors (TRAIL-R1-4) and expression levels of the antiapoptotic BCL-2 proteins MCL-1 and BCL-xL were analyzed by flow cytometry and Western blotting, respectively.Knock-down of MCL-1 and BCL-xL was performed by transfecting specific small interfering RNAs. HCC cells were treated with kinase inhibitors and chemotherapeutic drugs. Apoptosis induction and cell viability were analyzed via flow cytometry and 3-(4,5-Dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay.RESULTS: TRAIL-R1 and -R2 were profoundly expressed on the HCC cell lines Huh7 and Hep-G2.However, treatment of Huh7 and Hep-G2 with TRAIL and agonistic antibodies only induced minor apoptosis rates. Apoptosis resistance towards TRAIL could be considerably reduced by adding the chemotherapeutic drugs 5-fluorouracil and doxorubicin as well as the kinase inhibitors LY294002 [inhibition of phosphoinositol-3-kinase (PI3K)], AG1478 (epidermal growth factor receptor kinase), PD98059 (MEK1), rapamycin (mammalian target of rapamycin) and the multi-kinase inhibitor Sorafenib. Furthermore, the antiapoptotic BCL-2 proteins MCL-1 and BCL-xL play a major role in TRAIL resistance: knock-down by RNA interference increased TRAIL-induced apoptosis of H CC cells. Additionally,knock-down of MCL-1 and BCL-xL led to a significant sensitization of HCC cells towards inhibition of both c-Jun N-terminal kinase and PI3K.CONCLUSION: Our data identify the blockage of survival kinases, combination with chemotherapeutic drugs and targeting of antiapoptotic BCL-2 proteins as promising ways to overcome TRAIL resistance in HCC.     

siRNA沉默AFP基因对肝癌细胞系EGHC-9901凋亡的影响

目的建立稳定表达针对AFP基因siRNA质粒的肝癌细胞模型并探讨对其凋亡的影响。方法构建针对AFP基因的siRNAs表达质粒,脂质体法将该质粒转染肝癌细胞系EGHC-9901,G418筛选4~5周后Western blot及RT-PCR检测靶基因抑制效果,细胞分组：实验组（siRNA-afp）,转染AFP-siRNA质粒组;载体对照组,转染空载体组;空白对照组,未做任何处理组。免疫荧光检测细胞在无血清状态下凋亡情况,Western blot及RT-PCR检测Caspase-3、Caspase-8等凋亡相关蛋白表达。结果在体外成功构建针对AFP的siRNA表达质粒并在体外建立稳定表达该质粒肝癌细胞系EGHC-9901,转染后细胞表达AFP近乎完全抑制;免疫荧光表明促进细胞在无血清状态下的实验组凋亡率为32%±4%,对照组凋亡率为17%±3%,差异有统计学意义（P〈0.05）;RT-PCR及Western blot检测凋亡相关蛋白表明实验组Caspase-3表达较对照组高,差异有统计学意义（P〈0.05）,而Caspase-8、Caspase-9、bcl-2表达无显著差异（P〉0.05）。结论在体外成功建立稳定表达针对AFP基因的siRNA肝癌细胞系,抑制AFP表达可能通过上调Caspase-3表达促进其凋亡。     

PITX1和Pan-ras在肝癌细胞中的表达及意义

目的研究抑癌基因PITX1和其下游癌基因Pan-ras在正常胚肝细胞株L02，肝癌细胞株HepG2和SMMC-7721中的表达，探讨其在肝癌发生发展中的作用和关系。方法应用SABC免疫组织化学染色技术和westem blot蛋白质印迹以及半定量RT-PCR检测L02、HepG2和SMMC-7721细胞株中PITX1和Pan-ras基因的表达情况，并分析其意义。结果PITX1在肝癌细胞（HepG2、SMMC-7721）中表达比正常肝细胞L02显著降低，Pan-ras在肝癌细胞（HepG2、SMMC-7721）中的表达与正常肝细胞L02相比显著升高。结论PITX1在肝癌细胞中的低表达，以及Pan-ras的高表达，可能导致肝癌无限增殖，构成了肝癌细胞信号传导网络中的重要一环。     

Dual targeting of Polo-like kinase 1 and baculoviral inhibitor of apoptosis repeat-containing 5 in TP53-mutated hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma(HCC), often diagnosed at advanced stages without curative therapies, is the fifth most common malignant cancer and the second leading cause of cancer-related mortality. Polo-like kinase 1(PLK1) is activated in the late G2 phase of the cell cycle and is required for entry to mitosis. Interestingly, PLK1 is overexpressed in many HCC patients and is highly associated with poor clinical outcome. Baculoviral inhibitor of apoptosis repeatcontaining 5(BIRC5) is also highly overexpressed in HCC and plays key roles in this malignancy.AIM To determine the expression patterns of PLK1 and BIRC5, as well as their correlation with p53 mutation status and patient clinical outcome.METHODS The expression patterns of PLK1 and BIRC5, and their correlation with p53 mutation status or patient clinical outcome were analyzed using a TCGA HCC dataset. Cell viability, cell apoptosis, and cell cycle arrest assays were conducted to investigate the efficacy of the PLK1 inhibitors volasertib and GSK461364 and the BIRC5 inhibitor YM155, alone or in combination. The in vivo efficacy of volasertib and YM155, alone or in combination, was assessed in p53-mutated Huh7-derived xenograft models in immune-deficient NSIG mice.RESULTS Our bioinformatics analysis using a TCGA HCC dataset revealed that PLK1 and BIRC5 were overexpressed in the same patient subset and their expression was highly correlated. The overexpression of both PLK1 and BIRC5 was more frequently detected in HCC with p53 mutations. High PLK1 or BIRC5 expression significantly correlated with poor clinical outcome. PLK1 inhibitors(volasertib and GSK461364) or a BIRC5 inhibitor(YM155) selectively targeted Huh7 cells with mutated p53, but not HepG2 cells with wild-type p53. The combination treatment of volasertib and YM155 synergistically inhibited the viability of Huh7 cells via apoptotic pathway. The efficacy of volasertib and YM155, alone or in combination, was validated in vivo in a Huh7-derived xenograft model.CONCLUSION PLK1 and BIRC5 are highly co-expressed in p53-mutated HCC and inhibition of both PLK1 and BIRC5 synergistically compromises the viability of p53-mutated HCC cells in vitro and in vivo.     

肝细胞肝癌谷胱甘肽硫转移酶P1甲基化研究

目的本研究探讨谷胱甘肽硫转移酶（GST）PI启动子CpG岛甲基化与肝细胞肝癌（HCC）的关系。方法用甲基化特异性PCR技术检测53例HCC肿瘤组织及其癌旁非肿瘤组织中，GSTP1基因启动子CpG岛甲基化状况。结果GSTP1基因在HCC肿瘤组织中的甲基化率显著高于癌旁非肿瘤组织（X^2=19．08，P〈0．001），在Ⅲ-Ⅳ期肿瘤中的甲基化率显著高于Ⅰ-Ⅱ期肿瘤（X^2=4．84，P=0．028），在不同大小的肿瘤之间、单个结节与多个结节的肿瘤之间以及包膜完整的肿瘤与包膜不完整的肿瘤之间甲基化率的差异均无显著性。结论GSTP1启动子CDG岛甲基化可能参与肝细胞的痛性转变。     

Clinicopathological significance of RASSF1A reduced expression and hypermethylation in hepatocellular carcinoma

Purpose  

Protein downregulation and hypermethylation of Ras association domain family 1A (RASSF1A) has been recognized as an important early event in different classes of carcinogenesis, but clinicopathological significance of RASSF1A protein expression and methylation in hepatocellular carcinoma (HCC) remains largely unknown. The aim of the study was to investigate the expression of RASSF1A protein and methylation in HCC and their clinical significance.     

RhoC和IQGAP1蛋白在原发性肝细胞癌中的表达及意义

目的:探讨RhoC和IQGAP1蛋白在原发性肝细胞癌(hepatocellular carcinoma,HCC)组织中的表达及其临床病理意义.方法:采用免疫组织化学法分别检测56例原发性肝细胞癌和15例正常肝组织中RhoC及IQGAP1蛋白的表达,并分析两者的相关性及与临床病理因素的关系.结果:RhoC和IQGAP1在...     

lncRNA-SNHG15 accelerates the development of hepatocellular carcinoma by targeting mi R-490-3p/histone deacetylase 2 axis

BACKGROUND Hepatocellular carcinoma(HCC) has become a great threat for people's health.Many long noncoding RNAs are involved in the pathogenesis of HCC.SNHG15,as a tissue specific long noncoding RNAs,has been studied in many human cancers,except HCC.AIM To explore the regulatory mechanism of SNHG15 in HCC METHODS In the present research,101 HCC patient samples,two HCC cell lines and one normal liver cell line were used.RT-qPCR and Western blot analysis were applied to detect SNHG15,miR-490-3 p and histone deacetylase 2(HD AC2)expression.The regulatory mechanism of SNHG15 was investigated using CCK8,Transwell and luciferase reporter assays.RESULTS Our research showed that up-regulation of SNHG15 was found in HCC and was related to aggressive behaviors in HCC patients.Moreover,knockdown of SNHG15 restrained HCC cell proliferation,migration and invasion.In addition,SNHG15 served as a molecular sponge for miR-490-3 p.Further,miR-490-3 p directly targets HDAC2.HD AC2 was involved in HCC progression by interacting with the SNHG15/miR-490-3 p axis.CONCLUSION In conclusion,long noncoding RNA SNHG15 promotes HCC progression by mediating the miR-490-3 p/HDAC2 axis in HCC.     

锌α2糖蛋白对实验性大鼠肝癌的抑制作用

目的探讨锌α2糖蛋白(AZhGP1)与肝细胞癌(HCC)发生、发展和转移的关系。方法二乙基亚硝胺(DEN)构建肝硬化和HCC大鼠模型,以过表达AZGP1基因干扰的HepG2细胞液构建裸鼠皮下成瘤和肝原位移植模型。免疫组织化学、蛋白免疫和PCR检测AZGP1和TGFβ1的表达。结果AZGP1 mRNA在正常肝组织、肝硬化和肝癌中的表达分别为(0.98±0.02)、(0.52±0.03)、(0.20±0.02);而TGFβ1的基因和蛋白表达在肝硬化和肝癌组织中呈现显著上调。AZGP1过表达的HepG2细胞接种裸鼠构建皮下肿瘤(HepG2-AZGP1组)。与对照组(HepG2-GFP组)比较,肿瘤大小无差异。裸鼠肝内移植术6周后,HepG2-GFP组57%和HepG2-AZGP1组14%发生肺转移(P=0.0157)。与HepG2-GFP组比较,HepG2-AZGP1组肝脏原发灶和肺转移灶结节数目明显减少,癌细胞异型性明显较轻。结论肝硬化进展至肝癌过程中抑癌基因.AZGP1发生缺失,伴随着失去阻抑TGFβ1作用。恢复AZGP1功能可能是一种新的有前途的治疗肝癌方法。