Sorafenib for recurrence of hepatocellular carcinoma after liver transplantation

BackgroundRecurrence of hepatocellular carcinoma after orthotopic liver transplantation not amenable to surgical approaches is associated with poor outcome.AimsRetrospective evaluation of the safety and efficacy of sorafenib in patients with post-transplant hepatocellular carcinoma recurrence.MethodsPatients with post-transplant hepatocellular carcinoma recurrence were treated with sorafenib. Adverse events were assessed using National Cancer Institute Common Toxicity Criteria of AEs version 3.0, tumour response was evaluated according to Response Evaluation Criteria in Solid Tumours.ResultsFirst-line therapy after recurrence was surgery (n = 6), radiation therapy (n = 1), chemotherapy (n = 1), and sorafenib (n = 3). Finally, 11 patients were treated with sorafenib. Nine patients (82%) received an additionally targeted therapy with sirolimus as part of their immunosuppressive regimen. Most common grade 3 adverse events included diarrhoea (46%), hand-foot skin reaction (27%), nausea, fatigue, and leucopoenia (all 18%). Sorafenib had to be discontinued in two patients due to adverse events and six additional patients required a dose adjustment. No deterioration of liver graft function occurred. Median time to progression was 4.1 months; however, patients were treated with ongoing sorafenib in case of clinical benefit (median 8.9 months). Median overall survival after initiation of sorafenib treatment was 20.1 months.ConclusionSorafenib in combination with immunosuppression including sirolimus may be administered to patients with post-transplant hepatocellular carcinoma recurrence with acceptable toxicity and without deterioration of liver graft function.     

肝癌转移、复发预测的蛋白质分子标志物

肝癌转移是一个多步骤、多因素相互作用的复杂过程.在肝癌转移复发的蛋白质组学研究方面,发现了很多新的潜在蛋白质分子标志物.由于仅用单个指标是无法进行准确预测的,采用多个指标进行联合检测将是今后肝癌转移预测的研究方向.本文综述了所涉及肝癌转移复发早诊的新型分子标志物.     

单克隆靶向转移人肝癌细胞株和裸鼠移植模型的建立

目的 建立具有器官转移亲嗜性的单克隆人肝癌细胞株和相应的裸鼠移植模型. 方法 取肺和淋巴结转移亲嗜性人肝癌荧光细胞株HCCLM3-R-LM1及HCCLM3-R-LnM1,通过极限稀释法进行单细胞培养,获得8个HCCLM3-R-LM1来源的单克隆细胞株(LM1-S2,-S3,-S4,-S5,-S11,-S15,-S21,-S23)和5个HCCLM3-R-LnM1来源的单克隆细胞株(LnM1-S7,-S11,-S13,-S17,-S20);将上述人肝癌单克隆细胞分别接种于4周龄的裸鼠皮下,3周后皮下瘤组织移植至裸鼠的肝脏,6周后观察裸鼠肺和腹腔淋巴结转移灶荧光面积,并与肺组织连续切片中的转移灶数目进行比较.同一株细胞的肺和淋巴结转移情况比较用Wilcoxon秩和检验,单克隆细胞株之间的肺、淋巴结转移情况比较用Kruskal-Wallis检验. 结果 在13株人肝癌单克隆细胞中,有6株细胞形成皮下瘤;移植至裸鼠肝脏后表现出不同的转移潜能和器官靶向特性.其中LM1-S3,LM1-S4,LM1-S5和LM1-S11单克隆细胞的肺转移积分吸光度值分别为80923±10162、1506000±297064、36 140±8210和508448±1342729(P＜0.01),但不发生淋巴结转移.LnM1-S11单克隆细胞的肺与淋巴结转移灶积分吸光度值分别为435 062±206 620和1254000±225171. 结论 成功建成了不同转移潜能和器官亲嗜性的人肝癌单克隆细胞株和裸鼠移植模型,其中LM1-S3,LM1-S4,LM1-S5和LM1-S11为肺特异亲嗜性的人肝癌单克隆细胞株,而LnM1-S11细胞为肺和淋巴结双重亲嗜性的人肝癌单克隆细胞株,为肝癌转移器官靶向性研究提供了理想的体内外模型.     

Sorafenib in the treatment of hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the 5th most frequent tumour disease and, at the same time, the 3rd most frequent cause of death from cancer worldwide. More than 600 000 new patients are diagnosed every year and more than 80% are diagnosed at an advance stage where surgical treatment is not indicated and systemic chemotherapy does not provide longer survival time. Sorafenib is the first substance that provides proven significant prolongation of survival time of HCC patients. This is a multikinase inhibitor with anti-proliferative and anti-angiogenic properties. Its efficacy was shown in the SHARP study that enabled licensing of sorafenib for the therapy of inoperable, metastasizing hepatocellular carcinoma, including patients with liver cirrhosis, functional class Child-Pugh A or B, in more than 60 countries worldwide, including the Czech Republic. The aim of this paper is to provide a comprehensive summary of the current treatment of HCC and, at the same time, to point out some new therapeutic approaches that, in the near future, shall certainly play a major role in the treatment of HCC.     

Regression of established hepatocellular carcinoma is induced by chemoimmunotherapy in an orthotopic murine model

The high rate of mortality and frequent incidence of recurrence associated with hepatocellular carcinoma (HCC) reveal the need for new therapeutic approaches. In this study we evaluated the efficacy of a novel chemoimmunotherapeutic strategy to control HCC and investigated the underlying mechanism that increased the antitumor immune response. We developed a novel orthotopic mouse model of HCC through seeding of tumorigenic hepatocytes from SV40 T antigen (Tag) transgenic MTD2 mice into the livers of syngeneic C57BL/6 mice. These MTD2-derived hepatocytes form Tag-expressing HCC tumors specifically within the liver. This approach provides a platform to test therapeutic strategies and antigen-specific immune-directed therapy in an immunocompetent murine model. Using this model we tested the efficacy of a combination of oral sunitinib, a small molecule multitargeted receptor tyrosine kinase (RTK) inhibitor, and adoptive transfer of tumor antigen-specific CD8(+) T cells to eliminate HCC. Sunitinib treatment alone promoted a transient reduction in tumor size. Sunitinib treatment combined with adoptive transfer of tumor antigen-specific CD8(+) T cells led to elimination of established tumors without recurrence. In vitro studies revealed that HCC growth was inhibited through suppression of STAT3 signaling. In addition, sunitinib treatment of tumor-bearing mice was associated with suppression of STAT3 and a block in T-cell tolerance. CONCLUSION: These findings indicate that sunitinib inhibits HCC tumor growth directly through the STAT3 pathway and prevents tumor antigen-specific CD8(+) T-cell tolerance, thus defining a synergistic chemoimmunotherapeutic approach for HCC.     

Sorafenib induced tumor lysis syndrome in an advanced hepatocellular carcinoma patient

A 55-year-old male patient with hepatitis B-related liver cirrhosis was found to have advanced hepatocellular carcinoma. His AFP was initially 9828 μg/L and rapidly dropped to 5597 μg/L in ten days after oral sorafenib treatment. However, he developed acute renal failure, hyperkalemia, and hyperuricemia 30 d after receiving the sorafenib treatment. Tumor lysis syndrome was suspected and intensive hemodialysis was performed. Despite intensive hemodialysis and other supportive therapy, he developed multiple organ failure （liver, renal, and respiratory failure） and metabolic acidosis. The patient expired 13 d after admission.     

Macrophages promote tumour growth and liver metastasis in an orthotopic syngeneic mouse model of colon cancer

Purpose

Tumour-associated macrophages have been shown to promote proliferation, angiogenesis and metastasis in several carcinomas. The effect on colon cancer has not yet been clarified. Furthermore, Kupffer cells in the liver might initiate the formation of metastases by directly binding tumour cells.

Methods

An orthotopic syngeneic mouse model of colon cancer as well as a liver metastases model has been studied, using murine CT-26 colon cancer cells in Balb/c-mice. Macrophages were depleted in both models by clodronate liposomes. Tumour sizes and metastases were determined using 7-Tesla MRI. The macrophage and vascular density in the orthotopic tumours as well as the Kupffer cell density in the livers were evaluated using immunohistochemistry.

Results

Animals in the macrophage-depleted group displayed significantly smaller primary tumours (37?±?20 mm³) compared to the control group (683?±?389 mm³, p?=?0.0072). None of the mice in the depleted group showed liver or peritoneal metastases, whereas four of six control mice displayed liver and five out of six mice peritoneal metastases. The vascular density was significantly lower in the macrophage-depleted group (p?=?0.0043). In the liver metastases model, animals of the Kupffer cell-depleted group (14.3?±?7.7) showed significantly less liver metastases than mice of the two control groups (PBS liposomes, 118.5?±?28.2, p?=?0.0117; NaCl, 81.7?±?23.2, p?=?0.0266). The number of liver metastases correlated directly with the Kupffer cell density (p?=?0.0221).

Conclusion

Macrophages promote tumour growth, angiogenesis and metastases in this orthotopic syngeneic mouse model. Kupffer cells enhance the formation of metastases in the liver.     

Recent progress in molecular mechanisms of postoperative recurrence and metastasis of hepatocellular carcinoma

Hepatectomy is currently considered the most effective option for treating patients with early and intermediate hepatocellular carcinoma (HCC). Unfortunately, the postoperative prognosis of patients with HCC remains unsatisfactory, predominantly because of high postoperative metastasis and recurrence rates. Therefore, research on the molecular mechanisms of postoperative HCC metastasis and recurrence will help develop effective intervention measures to prevent or delay HCC metastasis and recurrence and to improve the long-term survival of HCC patients. Herein, we review the latest research progress on the molecular mechanisms underlying postoperative HCC metastasis and recurrence to lay a foundation for improving the understanding of HCC metastasis and recurrence and for developing more precise prevention and intervention strategies.     

Targeting of circulating hepatocellular carcinoma cells to prevent postoperative recurrence and metastasis

Currently,the main treatment for hepatocellular carcinoma(HCC)involves the surgical removal of tumors or liver transplantation.However,these treatments are often not completely curative,as they are associated with a risk for postoperative recurrence and metastasis.Circulating tumor cells(CTCs)are increasingly recognized as the main source for recurrence and metastasis after radical hepatectomies are performed.Many studies have demonstrated the association between the presence of either pre-or postoperative CTCs and an increased risk for HCC recurrence.To improve the therapeutic outcome of HCC,a personalized,comprehensive and multidisciplinary approach should be considered,involving the application of appropriate diagnostic and therapeutic measures targeting HCC CTCs in different stages throughout the course of treatment.This article proposes some HCC CTC-based strategies for the treatment of HCC,including the monitoring of HCC CTCs before,during and after radical hepatectomy,therapeutic targeting of HCC CTCs,prevention of the generation and colonization of CTCs,as well as the use of CTC indexes for the selection of indications,prediction of prognoses,and planning of individualized therapeutic regimens.Innovation and technological development of therapies targeting CTCs,as well as their translation into clinical practice,will help to effectively reduce postoperative recurrence and metastasis,and significantly prolong the survival of HCC patients.     

人肝癌肺和淋巴结靶向转移细胞及裸鼠移植模型的建立

目的 克隆具有肺和淋巴结转移亲嗜性的人肝癌细胞并建立相应的裸鼠移植模型.方法 将荧光人肝癌细胞HCCLM3-R接种于4周龄的裸鼠肝脏,6周后在荧光解剖下观察并获取肺及腹腔淋巴结转移灶组织,经体外克隆培养,所获细胞分别标记为HCCLM3-R-LM1和HCCLM3-R-LnM1.接种上述两种细胞于4周龄裸鼠肝脏,观察各自肺和腹腔淋巴结转移灶荧光数量,并与肺组织连续切片中的转移灶数目进行比较.计量资料采用Wilcoxon秩和检验和Kruskal-Wallis秩和检验进行统计学分析.结果 HCCLM3-R-LM1、HCCLM3-R和HCCLM3-R-LnM1细胞接种于裸鼠肝脏后第6周,在肺和腹腔淋巴结中均能发现肿瘤转移.3株细胞接种后,裸鼠肺和腹腔淋巴结转移灶荧光面积分别为8687.00±1844.63和2570.00±318.20(P＜0.001),6457.67±832.62和10 994.33±2212.31(P＜0.001),2968.67±2571.00和24 416.00±7186.13(P＜0.001),每只裸鼠光镜下肺转移灶中位数分别为755、430、310个(P＜0.001),与荧光定量结果相吻合.结论 成功建成人肝癌肺和淋巴结亲嗜性转移细胞和裸鼠移植模型,其中HCCLM3-R-LM1细胞具有明显的肺转移特性,而HCCLM3-R-LnM1细胞具有明显的淋巴结转移特性,为肝癌器官靶向转移研究提供了理想的体内外模型.

Abstract：

Objective To establish a systematic site-specific metastatsis model of human hepatocellular carcinoma (HCC) in nude mouse.Methods HCCLM3-R cells were seeded into mice liver to establish xenograft mouse models.With the help of RFP,metastasis foci in lungs and lymph nodes in mice were detected using fluorescent stereomicroscopy and were removed.Cells derived from the metastasis foci were named HCCLM3-R-LM1 and HCCLM3-R-LnM1 respectively.HCCLM3-R-LM1 and HCCLM3-R-LnM1 cells were seeded into mice livers to analyze the lung and lymph node metastasis.Lungs of all tested mice were collected,examined by pathological evaluation and counted lung metastasis.Results Both lung and lymph node metastasis were found in HCCLM3-R-LM1,HCCLM3-R and HCCLM3-R-LnM1 cells and a significant difference was found between the lung and the lymph node metastasis levels in the three cells.The fluorescent areas (pixels) of lung and lymph node metastasis were 8687.00 ± 1844.63 versus 2570.00 ±318.20 (P = 0.0031) in HCCLM3-R-LM1 cells,6457.67±832.62 versus 10 994.33±2 212.31(P=0.0036) in HCCLM3-R cells,and 2968.67 ± 2571.00 versus 24 416.00 ± 7 186.13 (P = 0.0094) in HCCLM3-R-LnM 1 cells,respectively.The middle numbers of microscopic lung metastatic foci were 775,430 and 310in HCCLM3-R-LM1,HCCLM3-R and HCCLM3-R-LnM1 cells (P＜0.001),respectively,consist with the results quantified by RFP.Conclusion We established the systematic site-specific metastasis models which demonstrates lung- and lymph node-specific metastasis potential in nude mice and can be used as a model for researches on site-specific metastasis of HCC.     

Expression of X-linked inhibitor-of-apoptosis protein in hepatocellular carcinoma promotes metastasis and tumor recurrence

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. Despite significantly improved diagnosis and treatment in recent years, the long-term therapeutic effect is compromised by the frequent recurrence and metastasis, of which the molecular mechanisms are not fully understood. Our initial studies in established HCC cell lines with different metastatic capabilities indicated a correlation of metastasis with the resistance to apoptosis and therefore the ability to survive in stressed conditions. Subsequent investigation revealed that increased expression of X-linked inhibitor-of-apoptosis protein (XIAP) was correlated with the resistance to apoptosis and enhanced invasiveness in vitro, which could contribute to increased metastatic foci in vivo. Furthermore, we found that nearly 90% of clinical samples from advanced HCC patients expressed high levels of XIAP. Patients with XIAP-positive tumors had a significantly increased risk of relapse, which resulted from metastasis after total liver resection and orthotopic liver transplantation. Indeed, XIAP expression could be an independent prognostic factor for predicting disease-free survival rate and overall survival rate of these patients. XIAP expression was also highly correlated with advanced cases that exceeded the Milan criteria and could be a prognostic factor for disease-free survival in these patients as well. CONCLUSION: Our studies have shown an important molecule in controlling HCC metastasis, defined a biomarker that can be used to predict HCC recurrence and patient survival after treatment, and suggest that XIAP can be a molecular target subject to intervention to reduce metastasis and recurrence.     

Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma

In our previous study, osteopontin (OPN) was identified as one of the leading genes that promote the metastasis of hepatocellular carcinoma (HCC). However, the mechanism by which OPN promotes metastasis of HCC is not understood. In this study, RNA interference mediated by viral vectors-which could induce a long-lasting down-regulation in gene expression-was applied to analyze the role of OPN in metastasis of HCC. Three lentiviral vectors encoding microRNA against OPN, Lenti.OPNi-1, Lenti.OPNi-2, and Lenti.OPNi-3, were constructed and found to down-regulate the OPN level by 62%, 78%, and 95%, respectively, in HCCLM3 cells which had an overexpression of OPN and a higher metastatic potential. Consequently, both Lenti.OPNi-2 and Lenti.OPNi-3 induced a significant decrease in matrix metalloproteinase (MMP)-2 and urokinase plasminogen activator expression, and led to an obvious inhibition of both in vitro invasion and in vivo lung metastasis of HCCLM3 cells (P < 0.001). Moreover, Lenti.OPNi-3, rather than Lenti.OPNi-2, could also suppress in vitro proliferation and in vivo tumor growth of HCCLM3. Smaller detectable tumors were found in only 50% of mice after implantation of Lenti.OPNi-3-transfected HCCLM3 cells (341 +/- 502.6 mm(3) versus >3500 mm(3) in controls; P < 0.001). Lenti.OPNi-3, not Lenti.OPNi-2, significantly suppressed the MEK/ERK1/2 pathway in HCCLM3 cells. Recombinant OPN was found to induce translocation of p65 into the nucleus of HCC cells and activation of MMP-2 and MEK/ERK/1/2, which were suppressed by the nuclear factor kappaB (NF-kappaB) inhibitor pyrrolidine dithiocarbamate. Conclusion: OPN plays an important role in metastasis as well as tumor growth of HCC, in which different minimum threshold levels of OPN are needed. These effects may occur through activation of the mitogen-activated protein kinase and NF-kappaB pathways, and MMP-2. OPN could be a hopeful target for the control of HCC.     

Targeting vascular endothelial growth factor with the monoclonal antibody bevacizumab inhibits human hepatocellular carcinoma cells growing in an orthotopic mouse model

Background: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer related death worldwide. Liver resection or transplantation is curative for a subset of patients with localized disease, but treatments for advanced disease are generally toxic and ineffective. Aberrant expression of the vascular endothelial growth factor (VEGF) has been implicated in the progression of HCC and represents a valid target for anticancer therapy. Bevacizumab, a humanized anti‐VEGF monoclonal antibody, is currently being evaluated in the treatment of HCC. In addition, other novel anti‐angiogenesis agents are being developed in HCC. Aim: This study examines the effect of bevacizumab in a newly characterized orthotopic model of the disease using the human HCC cell line, Hep 3B, and provides preclinical evidence that an anti‐angiogenic approach holds promise in HCC. Results: Administration of bevacizumab 5 mg/kg intraperitoneal twice a week significantly decreased microvessel density in tumours, decreased human serum α‐fetoprotein measurements and prolonged the time to progression for treatment mice compared with control mice. Conclusions: Our findings suggest that targeting VEGF with bevacizumab may be an effective approach to the treatment of HCC and further study of other novel anti‐angiogenic agents in HCC is warranted.