Inhibition of histone deacetylase for the treatment of biliary tract cancer： A new effective pharmacological approach

AIM:TO investigate in vitro and in vivo therapeutic effects of histone deacetylase inhibitors NVP-LAQ824 and NVP-LBH589 on biliary tract cancer. METHODS:Cell growth inhibition by NVP-LAQ824 and NVP-LBH589 was studied in vitro in 7 human biliary tract cancer cell lines by MTT assay.In addition,the anti- tumoral effect of NVP-LBH589 was studied in a chimeric mouse model.Anti-tumoral drug mechanism was assessed by immunoblotting for acH4 and p21~(WAF-1/CIP-1), PARP assay,cell cycle analysis,TUNEL assay,and immunhistochemistry for MIB-1. RESULTS:In vitro treatment with both compounds significantly suppressed the growth of all cancer cell lines[mean IC_(50)(3 d)0.11 and 0.05μmol/L, respectively],and was associated with hyperacetylation of nucleosomal histone H4,increased expression of p21~(WAF-1/CIP-1),induction of apoptosis(PARP cleavage),and cell cycle arrest at G2/M checkpoint.After 28 d,NVP- LBH589 significantly reduced tumor mass by 66%(bile duct cancer)and 87%(gallbladder cancer)in vivo in comparison to placebo,and potentiated the efficacy of gemcitabine.Further analysis of the tumor specimens revealed increased apoptosis by TUNEL assay and reduced cell proliferation(MIB-1). CONCLUSION:Our findings suggest that NVP-LBH589 and NVP-LAQ824 are active against human biliary tract cancer in vitro.In addition,NVP-LBH589 demonstrated significant in vivo activity and potentiated the efficacy of gemcitabine.Therefore,further clinical evaluation of this new drug for the treatment of biliary tract cancer is recommended.     

Molecular genetics and targeted therapeutics in biliary tract carcinoma

The primary malignancies of the biliary tract, cholangio-carcinoma and gallbladder cancer, often present at an advanced stage and are marginally sensitive to radiation and chemotherapy. Accumulating evidence indicates that molecularly targeted agents may provide new hope for improving treatment response in biliary tract carcinoma(BTC). In this article, we provide a critical review of the pathogenesis and genetic abnormalities of biliary tract neoplasms, in addition to discussing the current and emerging targeted therapeutics in BTC. Genetic studies of biliary tumors have identified the growth factors and receptors as well as their downstream signaling pathways that control the growth and survival of biliary epithelia. Target-specific monoclonal antibodies and small molecules inhibitors directed against the signaling pathways that drive BTC growth and invasion have been developed. Numerous clinical trials designed to test these agents as either monotherapy or in combination with conventional chemotherapy have been completed or are currently underway. Research focusing on understanding the molecular basis of biliary tumorigenesis will continue to identify for targeted therapy the key mutations that drive growth and invasion of biliary neoplasms. Additional strategies that have emerged for treating this malignant disease include targeting the epigenetic alterations of BTC and immunotherapy. By integrating targeted therapy with molecular profiles of biliary tumor, we hope to provide precision treatment for patients with malignant diseases of the biliary tract.     

Inhibitor of differentiation proteins do not influence prognosis of biliary tract cancer

AIM:To investigate the expression and clinical relevance of inhibitor of differentiation(ID)proteins in biliary tract cancer.METHODS:ID protein expression was analyzed in129 samples from patients with advanced biliary tract cancer(BTC)(45 extrahepatic,50 intrahepatic,and 34 gallbladder cancers),compared to normal controls and correlated with clinical an pathological parameters.RESULTS:ID1-3 proteins are frequently overexpressed in all BTC subtypes analyzed.No correlation between increased ID protein expression and tumor grading,tumor subtype or treatment response was detected.Survival was influenced primary tumor localization(extrahepatic vs intrahepatic and gall bladder cancer,OS 1.5 years vs 0.9 years vs 0.7 years,P=0.002),by stage at diagnosis(OS 2.7 years in stageⅠv s 0.6 years in stageⅣ,P<0.001),resection status and response to systemic chemotherapy.In a multivariate model,ID protein expression did not correlate with clinical prognosis.Nevertheless,there was a trend of shorter OS in patients with loss of cytoplasmic ID4 protein expression(P=0.076).CONCLUSION:ID protein expression is frequently deregulated in BTC but does not influence clinical prognosis.Their usefulness as prognostic biomarkers in BTC is very limited.     

EGFR and HER2 expression in advanced biliary tract cancer

AIM： To analyze the pathogenetic role and potential clinical usefulness of the epidermal growth factor receptor （EGFR） and the human epidermal growth factor receptor 2 （HER2） in patients with advanced biliary tract cancer （BTC）. METHODS： EGFR and HER2 expression was studied in biopsy samples from 124 patients （51% women; median age 64.8 years）, with advanced BTC diagnosed between 1997 and 2004. Five micrometers sections of paraffin embedded tissue were examined by standard, FDA approved immunohistochemistry. Tumors with scores of 2＋ or 3＋ for HER2 expression on immunochemistry were additionally tested for HER2 gene amplification by fluorescence in situ hybridisation （FISH）.RESULTS： 34/124 patients （27.4%） had gallbladder cancer, 47 （37.9%） had intrahepatic BTC and 43 （34.7%） had extrahepatic or perihilar BTC. EGFR expression was examined in a subset of 56 samples. EGFR expression was absent in 22/56 tumors （39.3%）. Of the remaining samples expression was scored as 1＋ in 12 （21.5%）, 2＋ in 13 （23.2%） and 3＋ in 9 （16%）, respectively. HER2 expression was as follows： score 0 73/124 （58.8%）, score 1＋ 27/124 （21.8%）, score 2＋ 21/124 （17%） and score 3＋ 4/124 （3.2%）. HER2 gene amplification was present in 6/124, resulting in an overall amplification rate of 5%. CONCLUSION： Our data suggest that routine testing and therapeutic targeting of HER2 does not seem to be useful in patients with BTC, while targeting EGFR may be promising.     

Establishment of various biliary tract carcinoma cell lines and xenograft models for appropriate preclinical studies

We recently reported several driver genes of biliary tract carcinoma(BTC) that are known to play important roles in oncogenesis and disease progression. Although the need for developing novel therapeutic strategies is increasing, there are very few BTC cell lines and xenograft models currently available for conducting preclinical studies. Using a total of 88 surgical BTC specimens and 536 immunodeficient mice, 28 xenograft models and 13 new BTC cell lines, including subtypes, were established. Some of our cell lines were found to be resistant to gemcitabine, which is currently the first choice of treatment, thereby allowing highly practical preclinical studies to be conducted. Using the aforementioned cell lines and xenograft models and a clinical pathological database of patients undergoing BTC resection, we can establish a preclinical study system and appropriate parameters for drug efficacy studies to explore new biomarkers for practical applications in the future studies.     

KML001, an arsenic compound,as salvage chemotherapy in refractory biliary tract cancers: A prospective study

Background

Sodium meta-arsenite (NaAsO₂, KML001) is a potential oral anticancer agent acting on telomerase and telomere length. This prospective study evaluated its safety, tolerability, and effectiveness as salvage chemotherapy in patients with advanced biliary tract cancer (BTC) resistant to gemcitabine-based chemotherapy.

Improvement of prognosis for unresectable biliary tract cancer

AIM:To evaluate the chemotherapeutic outcomes and confirm the recent improvement of prognosis for unresectable biliary tract cancer.METHODS:A total of 186 consecutive patients with unresectable biliary tract cancer,who had been treated with chemotherapy between 2000 and 2009 at five institutions in Japan,were retrospectively analyzed.These patients were divided into three groups based on the year beginning chemotherapy:Group A(2000-2003),Group B(2004-2006),and Group C(2007-2009).The data were fixed at the end of December 2011.Overall survival and time-to-progression were analyzed and compared chronologically.RESULTS:No patient characteristics were significantly different among the three groups.The gallbladder was involved in about half of the patients in each group,and metastatic biliary tract cancer was present in three quarters of the enrollees.In Group A,5-fluorouracilbased chemotherapies were primarily selected as firstline chemotherapy,and only 24% were treated with second-line chemotherapy.In Group B,gemcitabine or S-1 monotherapy was mainly introduced as firstline chemotherapy,and 51% of the patients who were refractory to first-line chemotherapy were treated with second-line chemotherapy mainly with monotherapy.In Group C,the combination therapy with gemcitabine and S-1 was mainly chosen as first-line chemotherapy,and 53% of the patients refractory to first-line chemotherapy were treated with second-line chemotherapy mainly with combination therapy.The median timeto-progressions were 4.4 mo,3.5 mo and 5.9 mo in Groups A,B and C,respectively(4.4 mo vs 3.5 mo vs 5.9 mo,P < 0.01).The median overall survivals were 7.1,7.3,and 11.7 mo in Groups A,B and C(7.1 mo vs 7.3 mo vs 11.7 mo,P = 0.03).Induction rates of all three drugs(gemcitabine,platinum analogs,and fluoropyrimidine) in Groups A,B and C were 4%,2% and 27%(4% vs 2% vs 27%,P < 0.01).CONCLUSION:The prognosis of unresectable biliary tract cancer has improved recently.Using three effective drugs(gemcitabine,platinum analogs,and fluoropyrimidine) may     

A rare surgical case of metachronous double carcinoma of the biliary tract

We report on a rare case of metachronous double carcinoma of the biliary tract, occurring in a 65-year-old male. The patient was admitted to the hospital with jaundice in March 2004. Ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI) scans of the abdomen showed a minimally dilated intrahepatic biliary tree with normal-appearing choledocus. Obstruction of the common hepatic duct was revealed by endoscopic retrograde cholangiopancreatography (ERCP). The patient underwent a resection of the middle third of the extrahepatic duct and cholecystectomy (cholangiocarcinoma, pT1N0M0), with the surgical margins of resection showing as negative. After 2 years, during follow-up, the findings of a positron emission tomography (PET)-CT scan suggested a possible cholangiocarcinoma of the distal part of the biliary tract; CT and MRI scanning of the abdomen showed mild dilatation of the distal common hepatic duct; an ERCP showed mild dilatation of the retropancreatic remnant of the biliary tree with endoluminal defects. Eventually the patient underwent pancreaticoduodenectomy. The histopathological diagnosis of the resected specimen confirmed a cholangiocarcinoma; 10 lymph nodes were negative (pT1N0M0). At 6 months post-op after the second operation the patient is progressing well with no signs of recurrence. Patients with cholangiocarcinoma – in whom survival is prolonged with surgical resection – should undergo careful follow-up for both recurrence and second primary cancer. PET scanning seems to play the most important diagnostic role.     

曲古抑菌素A对乳腺癌细胞株MDA-MB-231增殖的影响

目的探讨曲古抑菌素A（TSA）对乳腺癌细胞株增殖的影响。方法培养乳腺癌细胞株MDA-MB-231,用组不同浓度TSA分别作用于细胞,于24、48、72、96h用MTT比色法观察细胞生长情况,流式细胞术检测S期细胞的比例和周期素（Cyclin）D1、A2表达。结果72h内和4μmol／L的浓度范围内,乳腺癌细胞生长受到抑制,并有一定的时效关系;TSA作用后周期素Cyclin A2表达下降,Cyclin D1表达上升（P均〈0．05）。结论TSA能抑制乳腺癌细胞生长,周期素Cyclin D1、Cyclin A2参与了抑制剂对细胞增殖周期的调控。     

Optimum chemotherapy for the management of advanced biliary tract cancer

Biliary tract cancers(BTCs) are highly fatal malignancies, which are often diagnosed at an advanced stage and have relatively poor prognosis.The treatment of patients with advanced BTC is systemic, based on chemotherapy or best supportive care, depending on their performance status.Despite clinical trials studyingmany chemotherapeutic regimens and targeted therapies for the treatment of BTC, the standard of care for advanced BTC remains the combination of gemcitabine with cisplatin.Many new molecules targeting proliferation and survival pathways, the immune response and angiogenesis are currently undergoing phase Ⅰ and Ⅱ trials for the treatment of advanced BTC with promising results.     

分子靶向治疗晚期胆系恶性肿瘤的研究进展

胆系恶性肿瘤(BTC)是起源于肝内外胆管及胆囊壁黏膜上皮细胞恶性肿瘤,约占消化系统恶性肿瘤的3％,近年来发病率和病死率不断增加.基于临床和病理异质性的特点,BTC早期发现困难,确诊时往往已处于晚期,预后极差.一直以来,晚期BTC都是唯化学治疗,但治疗效果不尽如人意.近年来,随着精准治疗的兴起,尤其是分子靶向药物在BTC...     

Treatment outcomes of chemotherapy between unresectable and recurrent biliary tract cancer

AIM: To evaluate the differences in the treatment outcomes between the unresectable and recurrent biliary tract cancer patients who received chemotherapy.METHODS: Patients who were treated with gemcitabine and S-1 combination therapy in the previous prospective studies were divided into groups of unresectable and recurrent cases. The tumor response, time-to-progression, overall survival, toxicity, and dose intensity were compared between these two groups.RESULTS: Response rate of the recurrent group was higher than that of the unresectable group (40.0% vs 25.5%; P = 0.34). Median time-to-progression of the recurrent and unresectable groups were 8.7 mo (95%CI), 1.2 mo, not reached) and 5.7 mo (95%CI: 4.0-7.0 mo), respectively (P = 0.14). Median overall survival of the recurrent and the unresectable groups were 16.1 mo (95%CI: 2.0 mo-not reached) and 9.6 mo (95%CI: 7.1-11.7 mo), respectively (P = 0.10). Dose intensities were significantly lower in the recurrent groups (gemcitabine: recurrent group 83.5% vs unresectable group 96.8%; P < 0.01, S-1: Recurrent group 75.9% vs unresectable group 91.8%; P < 0.01). Neutropenia occurred more frequently in recurrent group (recurrent group 90% vs unresectable group 55%; P = 0.04).CONCLUSION: Not only the efficacy but also the toxicity and dose intensity were significantly different between unresectable and recurrent biliary tract cancer.     

ERCP对肝外胆管癌的诊断价值

目的对手术病理证实的３１例肝外胆管癌进行回顾性分析．方法３１例患者均行胰胆管造影，并和Ｂ超及ＣＴ检查进行对照．结果上段胆管癌１６例，下段胆管癌１３例，中上段胆管癌２例．１例为横纹肌肉瘤，２例为腺瘤癌变，其余均为腺癌．ＥＲＣＰ成功２８例（９０３％），Ｂ超诊断符合率为８０７％，而ＣＴ显示梗阻８５０％，能明确病因者仅７００％．结论ＥＲＣＰ在早期诊断肝外胆管癌方面明显优于Ｂ超及ＣＴ检查，并能清晰地显示胆道系统的全貌，对治疗及手术方案选择有重要价值．     

组蛋白脱乙酰基酶抑制剂曲古霉素对心肌细胞肥大的影响

目的利用体外培养的大鼠乳鼠心肌细胞,研究曲古霉素(TSA)对血管紧张素II(Ang II)诱导的心肌细胞肥大的影响,并探讨相关机制。方法分离乳鼠心肌细胞进行原代培养,应用血管紧张素Ⅱ制备心肌细胞肥大模型。分别给予不同浓度的TSA预处理心肌细胞,观察TSA对心肌细胞面积、3H亮氨酸掺入率、心房利钠肽(ANP)、脑利钠肽(BNP)的mRNA表达水平的影响。同时通过检测乙酰化组蛋白3的蛋白表达水平,观察AngⅡ和TSA对组蛋白脱乙酰基酶(HDAC)活性的影响。应用Western blot检测AngⅡ和TSA处理后磷酸化的c-Jun氨基末端激酶(JNK)表达水平的变化。结果10-6mmol/L AngⅡ作用48 h后,心肌细胞面积增加至对照组的(1.63±0.46)倍(P<0.01),而10-7mmol/L和3×10-7mmol/L TSA预处理可以剂量依赖性的抑制AngⅡ引起的心肌细胞面积增大。TSA干预也阻断了AngⅡ引起的蛋白合成速率增加以及ANP和BNP的蛋白表达水平的增加(均为P<0.05)。AngⅡ刺激心肌细胞使乙酰化组蛋白3的蛋白表达水平降低,TSA可逆转这一效应。同时TSA抑制了AngⅡ介导的磷酸化JNK表达水平的升高。结论 TSA可显著抑制AngⅡ诱导的心肌细胞肥大和HDAC活性增加,可能通过抑制JNK的激活而发挥抑制心室肥厚的作用。     

Clinical results of intra-arterial adjuvant chemotherapy for prevention of liver metastasis following curative resection of biliary tract cancer

Background. This is a report on the clinical results of intra-arterial adjuvant chemotherapy in the prevention of liver metastasis following curative resection of biliary tract cancer. Methods. Nineteen patients of advanced biliary tract underwent a pathologically radical operation between 2001 and 2006 (8 M and 11 F; mean age 66.2 years). Intra-arterial adjuvant chemotherapy with CDDP and 5-FU was performed selectively for 9 patients. The control group comprised 10 patients. Age, gender, staging of the disease, resection of the portal vein, postoperative radiotherapy, histological liver invasion as demographics and clinical characteristics were compared between the two groups. Results. Demographics and clinical characteristics were similar in the two groups. Liver metastasis occurred in 4 of 9 patients (44.4%) in the chemotherapy group and in 5 of 10 patients (50%) in the control group. There was no difference in the rate of liver metastasis between the two groups. The median survival term was 23.3 months for 9 patients who underwent the intra-arterial adjuvant chemotherapy, whereas the median survival term for 10 patients who were curatively resected without intra-arterial adjuvant chemotherapy was 21.7 months. The median survival term was statistically similar in both groups. Furthermore, in the recurrence-free survival, there was no major difference between the chemotherapy and control groups statistically. Conclusions. In the patients with advanced biliary tract cancer who underwent the curative operation, the intra-arterial adjuvant chemotherapy could not suppress the rate of liver metastasis nor improve cumulative survival.