Low serum interleukin-6 levels as a predictive marker of recurrence in patients with hepatitis B virus related hepatocellular carcinoma who underwent curative treatment

BackgroundWe aimed to investigate the use of novel serum biomarkers for predicting the recurrence and survival of patients with hepatitis B virus (HBV)-related early hepatocellular carcinoma (HCC) after hepatic resection or radiofrequency ablation (RFA).MethodsOne hundred and five patients with HBV-related HCC, who fulfilled the Milan criteria without vascular invasion and underwent hepatic resection or RFA, were followed-up for a median duration of 52 months. Pretreatment serum concentrations of 16 cytokines including interleukin-6 (IL-6) were measured by using a Luminex 200 system. The measured serum cytokines and several clinical factors were analyzed retrospectively.ResultsUnivariate analysis showed that patients with lower pretreatment serum levels of IL-10, IL-6, monocyte chemoattractant protein-1, and tumor necrosis factor-α had significantly shorter disease-free survival (DFS) than those with higher levels. Multivariate analysis revealed that a low serum IL-6 level (⩽33.00 pg/mL; hazard ratio [HR] = 5.39; 95% confidence interval [CI] = 1.27–22.93; P = 0.022), low platelet count (<100 × 10⁹/L; HR = 2.23; 95% CI = 1.28–3.89; P = 0.005), and low serum albumin level (⩽3.5 g/L; HR = 2.26; 95% CI = 1.28–3.97; P = 0.005) had a negative prognostic impact on DFS. In the analysis for overall survival, a low serum platelet level (<100 × 10⁹/L; HR = 2.80; 95% CI = 1.31–5.99; P = 0.008) and multiple tumor (⩾2; HR = 4.05; 95% CI = 1.56–10.48; P = 0.004) showed a negative prognostic impact on the overall survival.ConclusionA low serum IL-6 level is, in addition to low platelet count and low serum albumin level, an independent prognostic factor for DFS in patients with HBV-related early HCC who underwent hepatic resection or RFA with curative intention.     

The Tumor Suppressor pVHL Down-regulates Never-in-Mitosis A-related Kinase 8 via Hypoxia-inducible Factors to Maintain Cilia in Human Renal Cancer Cells

NEK8 (never in mitosis gene A (NIMA)-related kinase 8) is involved in cytoskeleton, cilia, and DNA damage response/repair. Abnormal expression and/or dysfunction of NEK8 are related to cancer development and progression. However, the mechanisms that regulate NEK8 are not well declared. We demonstrated here that pVHL may be involved in regulating NEK8. We found that CAK-I cells with wild-type vhl expressed a lower level of NEK8 than the cells loss of vhl, such as 786-O, 769-P, and A-498 cells. Moreover, pVHL overexpression down-regulated the NEK8 protein in 786-O cells, whereas pVHL knockdown up-regulated NEK8 in CAK-I cells. In addition, we found that the positive hypoxia response elements (HREs) are located in the promoter of the nek8 sequence and hypoxia could induce nek8 expression in different cell types. Consistent with this, down-regulation of hypoxia-inducible factors α (HIF-1α or HIF-2α) by isoform-specific siRNA reduced the ability of hypoxia inducing nek8 expression. In vivo, NEK8 and HIF-1α expression were increased in kidneys of rats subjected to an experimental hypoxia model of ischemia and reperfusion. Furthermore, NEK8 siRNA transfection significantly blocked pVHL-knockdown-induced cilia disassembling, through impairing the pVHL-knockdown-up-regulated NEK8 expression. These results support that nek8 may be a novel hypoxia-inducible gene. In conclusion, our findings show that nek8 may be a new HIF target gene and pVHL can down-regulate NEK8 via HIFs to maintain the primary cilia structure in human renal cancer cells.     

一种“无血清”培养基对肿瘤细胞生长的支持

本文报告一种为适于杂交瘤细胞生长“无血清”培养基－适量的成牛血清低分子成分超滤液、１０ｍｇ／Ｌ转铁蛋白（Ｔ）、１０ｍｇ／Ｌ胰岛素（Ｉ）、２０μｍｏｌ／Ｌ乙醇胺（Ｅ）、４０ｎｍｏｌ／Ｌ硒酸钠（Ｓ）等诸补充成分替代胎牛血清加到基础液中－也完全适用于培养肿瘤细胞,且观察到与之相似的规律性结果,癌细胞在本“无血清”培养若的生长水平达到在含胎牛血清培养基中生长的水平。对于癌细胞的生长,ＬＭＷ－ＣＳ与Ｔ、Ｉ、     

Morphometric Analysis of Hepatocellular carcinoma

Ｍｏｒｐｈｏｍｅｔｒｉｃ　Ａｎａｌｙｓｉｓ　ｏｆ　Ｈｅｐａｔｏｃｅｌｌｕｌａｒ　ｃａｒｃｉｎｏｍａＬａｉＭａｏ－ｄｅ（来茂德）;ＣｈｅｎＰｅｉ－ｈｕｉ（陈培辉）ａｎｄＺｈｏｕＳｈｕｉ－ｙｕｎ（周水云）（ＤｅｐａｒｔｍｅｎｔｏｆＰａｔｈｏｌｏｇｙ,Ｚｈ...     

FSH通过SATB1调控上皮性卵巢癌ES-2细胞的增殖和侵袭活性

目的：探讨特异AT序列结合蛋白1（special AT-rich sequence-bindingprotein,SATB1）在卵泡刺激素（Follicle stimulating hor-mone,FSH）诱导的上皮性卵巢癌ES-2细胞增殖和侵袭中的作用。方法：以Real-timePCR检测不同浓度FSH（0、10、20、40、80mlU／ml）处理后sATB1基因mRNA表达水平的变化。实验分4组：①siCon组,转染si-阴性对照（si-Negativecontrol）序列的实验组,对SATBl无干扰作用;②siSATB1组：转染特异性干扰下调SATB1的siSATB1序列;（3）FSH＋siCon组：以FSH处理的siCon组;（4）FSH＋siSATBl组：以FSH处理的sisATB1组。MTT法检测4组细胞的增殖情况,Westernblotting技术检测4组细胞细胞周期蛋白（CyclinDl）,基质金属蛋白酶2（MMP．2）的蛋白表达情况,Transwell侵袭实验检测4组细胞侵袭能力的变化。结果：1．FSH＋siCon组的细胞增殖能力明显高于siCon组的细胞增殖能力,FSH＋siCon组的CyclinD1蛋白相对表达量0．90＋0．08明显高于siCon组的0．37＋0．01（P均〈0．01）,提示FSH具有促进ES．2细胞增殖的作用。2．FSH＋siCon组的穿膜细胞数（30212）个明显高于siCon组（13919）个,FSH＋siCon组的MMP．2蛋白相对表达量0．40＋0．01明显高于siCon组的0．28＋0．02,提示FSH具有促进ES．2细胞侵袭能力的作用。3．随着FsH浓度的增高,SATBlmRNA的表达量逐渐增加,分别为1,1．66±0．04,1．79±0．21,2.31±0．03,以FsH浓度为80mlU／m1时最显著（P〈0．05）。4．FSH＋siSATBl组的细胞增殖能力明显低于FSH＋siCon组的细胞增殖能力,FSH＋siSATBl组的CyclinD1蛋白相对表达量0．22±0．02明显低于FSH＋siCon组的0．90±0．08（P均〈0．01）;FSH＋siSATB1组的穿膜细胞数（5216）个低于FSH＋siCon组的（30212）个,FSH＋siSATB1组的MMP-2蛋白相对表达量0．15±0．00明显低于FSH＋siCon组的0．40±0．01（P均〈0．01）,FSH促进ES-2细胞增殖和侵袭的能力由于SATB1基因表达的下降而被阻断。结论：SATBl是FSH作用的重要靶分子,介导FSH对上皮性卵巢癌ES-2细胞系增殖、侵袭活性的调控。     

NEDD4 promotes cell growth and motility in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related death worldwide. In China, the situation is even worse as cancer incidence and mortality continue to increase rapidly. Although tremendous progress has been made toward HCC treatments, the benefits for liver cancer patients are still limited. Therefore, it is necessary to identify and develop novel therapeutic methods. Neuronally expressed developmentally downregulated 4 (NEDD4), an E3 ubiquitin ligase, plays a critical role in the development and progression of various types of human cancers. In our study, NEDD4 acts as an oncoprotein in both QGY7703 and SMMC7721 liver cancer cell lines. We found that depletion of NEDD4 by siRNA transfection led to inhibition of cell growth, invasion and migration, and promotion of apoptosis. In contrast, overexpression of NEDD4 via plasmid transfection resulted in facilitated cell proliferation, invasion and migration, and decreased apoptosis. Importantly, we observed that tumor suppressor LATS1, also a core component of Hippo pathway, was negatively regulated by NEDD4 in liver cancer cells. Our findings suggested that NEDD4 may be involved in the HCC progression via regulating LATS1 associated signaling pathway. Therefore, targeting NEDD4-LATS1 signaling could be a potential therapeutic option for HCC treatment.     

肺癌组织中岩藻糖化糖链结构免疫组化研究

Lewis X(Le~x)、唾液酸化的Lewis X(Sialyl Lewis X,SLe~x)和唾液酸化的双岩藻糖Lewis X(Sialyl Dimeric Lewis X,SDLe~x)是细胞表面外侧带α1,3岩藻糖的糖链结构。本文用免疫组化ABC法研究了肺癌原发灶、转移灶和癌旁组织中这三种抗原结构的表达。结果发现这三种抗原在肺癌细胞表面及胞浆中均有不同程度的表达,而在肺癌癌旁组织及正常肺组织中未见表达。有转移的肺癌和(或)低分化肺癌中这三种抗原结构的表达要明显高于未发生转移和高、中分化肺癌中相同抗原结构的表达。其中以SLe~x的表达与肺癌细胞的转移能力和分化程度关系最为密切。另外,肺癌浸润转移的淋巴结中也有Le~x、SLe~x的明显表达和SDLe~x的少量表达,而未被肺癌浸润转移的淋巴结中就没有它们的表达。     

Heat-shock protein 72 cell-surface expression on human lung carcinoma cells is associated with an increased sensitivity to lysis mediated by adherent natural killer cells

 The cell-surface expression patterns of major histocompatibility complex (MHC) class I, class II and heat-shock protein 72 (HSP72) molecules were measured on human lung (LX-1) and mammary (MX-1) carcinoma cells. No major differences were found in the MHC cell-surface expression pattern of both cell lines. However, they differ significantly in their capacity to express HSP72 on their cell surface. Under physiological conditions LX-1 cells express HSP72 molecules on more than 90% of the cells, whereas MX-1 cells exhibit no significant HSP72 cell-surface expression (less than 5%). These expression patterns remained stable in all further cell passages tested. The sensitivity to lysis mediated by an interleukin-2 (IL-2)-stimulated, adherent natural killer (NK) cell population could be correlated with the amount of cell-surface-expressed HSP72 molecules. By antibody-blocking studies, using HSP72-specific monoclonal antibody (mAb), a strong inhibition of lysis was only found with LX-1 cells but not with MX-1 cells. In contrast to the cell-surface expression, the cytoplasmic amount of HSP72 in MX-1 cells was twice as high compared to LX-1 cells under physiological conditions. After nonlethal heat-shock the rate of induction and the total cytoplasmic amounts of HSP72 were comparable in both cell lines. The clonogenic cell viability of LX-1 cells after incubation at temperatures ranging from 41°C to 44°C was significantly elevated compared to that of MX-1 cells. In conclusion we state the following: (i) HSP72 cell-surface expression on human carcinoma cells is independent of the cytoplasmic amount of HSP72; (ii) the cell-surface expression of HSP72 is associated with an increased sensitivity of tumour cells to lysis mediated by an IL-2-stimulated, adherent NK cell population; (iii) thermoresistance is not related to the cytoplasmic HSP72 level but might be related to the amount of HSP72 expressed on the cell surface. Received: 20 June 1996 / Accepted: 25 September 1996     

微小RNA(microRNA)在肝细胞肝癌发生发展中的作用研究进展

MicroRNA是近几年发现的一种非编码小分子RNA,其主要功能体现在参与调节生物个体发育、细胞增殖与分化、肿瘤细胞的发生等多种病理过程中。MicroRNA在调控癌基因和抑癌基因的表达过程中,以及在肝细胞肝癌发生和发展中发挥着关键性的作用。本综述对microRNA在肝细胞肝癌发生发展中的作用进行汇总,以期为肝细胞肝癌的临床诊断以及治疗提供理论参考。     

Discovery of small molecule antagonists of chemokine receptor CXCR6 that arrest tumor growth in SK-HEP-1 mouse xenografts as a model of hepatocellular carcinoma

The chemokine system plays an important role in mediating a proinflammatory microenvironment for tumor growth in hepatocellular carcinoma (HCC). The CXCR6 receptor and its natural ligand CXCL16 are expressed at high levels in HCC cell lines and tumor tissues and receptor expression correlates with increased neutrophils in these tissues contributing to poor prognosis in patients. Availability of pharmacologcal tools targeting the CXCR6/CXCL16 axis are needed to elucidate the mechanism whereby neutrophils are affected in the tumor environment. We report the discovery of a series of small molecules with an exo-[3.3.1]azabicyclononane core. Our lead compound 81 is a potent (EC₅₀ = 40 nM) and selective orally bioavailable small molecule antagonist of human CXCR6 receptor signaling that significantly decreases tumor growth in a 30-day mouse xenograft model of HCC.