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癌胚性TUFT1异常表达与HBV相关肝细胞癌的研究进展

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胃肠病学和肝病学杂志 2024年第3期33卷 342-346页

作者：姚敏巫梦娜赛文莉姚登福南通大学附属医院临床医学研究中心江苏南通226001 南通大学医学院免疫学系南通大学第二附属医院重症医学科

肝细胞癌(hepatocellular carcinoma,HCC)的预防、早期诊断及精准治疗仍是亟需攻克的难题。肝炎病毒(HBV or HCV)慢性持续性感染、化学致癌物摄入和非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)等与HCC进展密切相关。... 详细信息

肝细胞癌(hepatocellular carcinoma,HCC)的预防、早期诊断及精准治疗仍是亟需攻克的难题。肝炎病毒(HBV or HCV)慢性持续性感染、化学致癌物摄入和非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)等与HCC进展密切相关。最新发现,在牙釉质发育和矿化中起重要作用的釉丛蛋白(Tuftelin,TUFT1)异常转录,在人体组织中广泛分布。TUFT1作为酸性、磷酸化糖蛋白可促进肿瘤进展与转移。然而,TUFT1表达与HCC的研究报道尚不多见。本文综述了TUFT1基因结构、组织分布与肿瘤的关系,重点分析其在HBV-HCC进展中作用及其应用前景。

关键词： HBV-HCC 釉丛蛋白异常表达临床价值

Abnormal expression of insulin-like growth factor-Ⅱ and its dynamic quantitative analysis at different stages of hepatocellular carcinoma development

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Hepatobiliary & Pancreatic Diseases International 2008年第4期7卷 406-411页

作者： LiWei Qiu dengfu yao Lei Zong YuanYuan Lu Hua Huang Wei Wu XinHua Wu Research Center of Clinical Molecular Biology Affiliated Hospital of Nantong University Nantong 226001 China.

BACKGROUND:Hepatocellular carcinoma(HCC)is characterized by multiple causes,clear multiple stages and a multifocal process of tumor progression related intimately to the overexpression of many cellular factors. The aim of this study was to investigate the dynamic expression of insulin-like growth factor-Ⅱ(IGF-Ⅱ) and its abnormal alteration in the early stages of HCC development. METHODS:Hepatoma models were induced by 2-fluorenylacetamide(2-FAA)in male Sprague-Dawley *** changes of rat livers were assessed by pathological examination(HE staining).The levels of IGF-Ⅱexpression in the livers and sera of rats were quantitatively detected by an enzyme-linked immunosorbent assay(ELISA).Simultaneously,the expression and cellular distribution of liver IGF-Ⅱwere analyzed by immunohistochemistry. RESULTS:Histological examination confirmed that rat hepatocytes showed changes from granule-like degeneration to atypical hyperplasia to HCC,and progressively increasing hepatic IGF-Ⅱlevels during HCC *** levels of hepatic or serum IGF- Ⅱin HCC tissues and sera were significantly higher than those in normals and rats with *** immunohistochemical evidence indicated the positiveexpression and hepatocyte distribution of IGF-Ⅱin rat hepatoma.A positive relationship of IGF-Ⅱlevels was found between liver tissues and sera of experimental rats (P<0.01). CONCLUSION:Hepatic IGF-Ⅱmay participate in hepatocyte cancer development and detection of IGF-Ⅱ expression during HCC development could be a useful molecular marker for its early diagnosis.

关键词： hepatocellular carcinoma insulin-like growth factor-Ⅱ immunohistochemistry dynamic expression

癌胚性TUFT1过表达及基因转录干预对肝细胞癌的抑制作用

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胃肠病学和肝病学杂志 2024年第6期33卷 740-745页

作者：巫梦娜方蓉菲邰伯军谢群徐敏赛文莉姚登福姚敏南通大学附属医院临床医学研究中心江苏南通226001 南通大学医学院免疫学系南通大学第二附属医院重症医学科南通大学附属医院感染性疾病科海安市人民医院感染性疾病科

目的分析肝细胞癌(hepatocellular carcinoma,HCC)釉丛蛋白(tuftelin,TUFT1)表达及基因转录干预的抑制作用。方法收集术后HCC组织和癌旁组织,分析TUFT1表达;从不同肝癌细胞和对照肝LO2细胞株中提取TUFT1,以免疫印迹法筛选TUFT1低或过表... 详细信息

目的分析肝细胞癌(hepatocellular carcinoma,HCC)釉丛蛋白(tuftelin,TUFT1)表达及基因转录干预的抑制作用。方法收集术后HCC组织和癌旁组织,分析TUFT1表达;从不同肝癌细胞和对照肝LO2细胞株中提取TUFT1,以免疫印迹法筛选TUFT1低或过表达的癌细胞株;将已构建含TUFT1基因或干扰TUFT1-shRNA3的慢病毒质粒,分别转染低或过表达TUFT1的癌细胞株,以CCK-8法、划痕、Transwell法、流式细胞术等分析细胞增殖、侵袭、迁移等生物学行为;将稳定表达TUFT1的两种细胞株皮下接种裸鼠,分析对移植瘤生长的影响。结果肝癌组织TUFT1过表达,主要定位于细胞浆和细胞膜。肝癌组TUFT1阳性率为87.1%,均显著高于癌旁组(χ^(2)=18.563,P<0.001);肝癌细胞株中TUFT1表达明显高于LO2细胞(P<0.001)。体外研究,以有效的TUFT1-shRNA3质粒,转染高TUFT1表达的MHCC-97H细胞,在TUFT1功能丧失后癌细胞的增殖、侵袭和迁移能力显著抑制,凋亡率升高;以插入TUFT1基因的质粒,转染低TUFT1表达的Hep3B细胞,在TUFT1功能获得后癌细胞增殖、侵袭和迁移能力均显著增强。体内研究,将功能丧失的MHCC-97H细胞于裸鼠皮下接种,移植瘤中TUFT1下调,并显著抑制瘤体生长(Z=12.000,P<0.01)。结论TUFT1为癌胚性蛋白,对其干预有望成为潜在的HCC治疗的分子靶目标。

关键词：肝细胞癌釉丛蛋白转录干预生物学行为移植瘤

Quantitative analysis of hepatoma-specific α-fetoprotein(HS-AFP) by a new mini-column affinity chromatography and its clinical value in diagnosis of hepatocellular carcinoma

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The Chinese-German Journal of Clinical Oncology 2008年第3期7卷 131-134页

作者： Wei Wu dengfu yao Liwei Qiu Xiaoxiao Gu Xinhua Wu Research Center of Clinical Molecular Biology The Affiliated Hospital Nantong University Nantong 226001 China E-commerec Department School of Economy and Commercial Management Nanjing University of Chinese Medicine Nanjing 210046 China

Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin （LCA）-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay （RIA）. Results： Circulating AFP was separated into three peaks （AFP-1, AFP-2, and AFP-3） by LCA-affinity chromatography. Dunng the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HSAFP （AFP-3） from sera of HCC patients was eluted clearly on the LCA-sepharose gel mini-column with a solution containing α-methyI-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity （92.7%） and specificity （88.2%）. Conclusion： The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.

关键词： hepatoma-specific alpha-fetoprotein (HS-AFP) affinity chromatography hepatocellular carcinoma (HCC)

Abnormal expression of hypoxia inducible factor-1α and clinical values of molecular-targeted interference in hepatocellular carcinoma

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The Chinese-German Journal of Clinical Oncology 2012年第3期11卷 125-129页

作者： Shanshan Li dengfu yao Zhizhen Dong Yajie Qian Dandan Yu Ninghua yao Jie Chen Xiaodi Yan Chenglin Qin Oncology Center Affiliated Hospital of Nantong University Nantong 226001 China Research Center of Clinical Medicine Affiliated Hospital of Nantong University Nantong 226001 China Department of Diagnostics Affiliated Hospital of Nantong University Nantong 226001 China Medical College of Nantong University Nantong 226001 China

Objective:The aim of this study was to analyze the expression features of hypoxia inducible factor-1α (HIF-1α) in hepatocellular carcinoma (HCC) and effects of HIF-1α silencing on HepG2 ***:HIF-1α expression was analyzed in the self-control HCC specimens by *** HepG2 cells with miRNA transfection,the expression of HIF-1α was determined at mRNA or protein level by real-time polymerase chain reaction (PCR) or Western *** endothelial growth factor (VEGF) and angiopoietin-2 (ANG-2) were determined by *** of cell cycles and apoptosis of HepG2 cells were measured using a flow ***:Positive HIF-1α was brown and granule-like in the cytoplasm or *** difference was found between HCC (80%) and its surrounding tissues (100%,χ2=22.35,P < 0.001) and HIF-1α expression related to tumor *** 72 h after miRNA transfection,the expression of HIF-1α in HepG2 cells was down-regulated by 87% at mRNA or 65% at protein level,with VEGF and ANG-2 decreased to 54% and 36%,*** RNA interference combined with anti-cancer drug,the apoptotic rate of HepG2 cells was increasing from 22.46% ± 0.61% to 36.99% ± 0.88%,with up-regulation of G1 phase (65.68% ± 0.91%) and down-regulation of S phase (19.47 ± 1.34 %).Conclusion:Abnormal expression of HIF-1α is associated with development of HCC,and HIF-1α gene silencing can effectively inhibit HepG2 cell proliferation.

关键词： hepatocellular carcinoma (HCC) hypoxia inducible factor-1α (HIF-1α) expression RNA interference gene silencing

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肝细胞恶性转化与肝癌诊断的特异性标志物

中华肝脏病杂志 2011年第4期19卷 244-246页

作者：姚敏姚登福南通大学附属医院临床医学研究中心江苏省南通市226001

肝细胞癌（HCC）是由病毒、化学致癌物等多病因作用,因癌基因或癌相关基因激活、抗癌基因失活或胚胎期某些癌基因重新复活等诸多因素引起肝细胞生长失控而致癌变,其中数百种基因调控和表达与肝癌发生、发展相关.手术切除仍是肝癌治疗的... 详细信息

肝细胞癌（HCC）是由病毒、化学致癌物等多病因作用,因癌基因或癌相关基因激活、抗癌基因失活或胚胎期某些癌基因重新复活等诸多因素引起肝细胞生长失控而致癌变,其中数百种基因调控和表达与肝癌发生、发展相关.手术切除仍是肝癌治疗的首选方法,早诊断、早治疗是提高肝癌患者生存率的惟一途径.随着多种＂组学＂如基因组学、蛋白质组学、转录组学等新技术的发展和对肝癌发生机制研究的深入,越来越多的与肝细胞恶性转化相关分子标志物被发现,有望成为肝癌早期特异性诊断的潜在分子标志物.

关键词：癌,肝细胞诊断标志物

Diagnostic and monitoring values of circulating AFPmRNA and HS-GGT for hepatocellular carcinoma

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The Chinese-German Journal of Clinical Oncology 2014年第10期13卷 457-463页

作者： Li Wang Min yao yao yao Liuhong Pan Shaolin Lu dengfu yao Research Center of Clinical Medicine Affiliated Hospital of Nantong University Medical School of Nantong University The Hospital of Nantong Maternal and Child Care Service Center of Oncology Affiliated Hospital of Nantong University

Objective: Early finding of hepatocellular carcinoma(HCC) and monitoring of its metastasis are of the utmost importance. The objectives of this study were to investigate the values of circulating hepatoma-specific gamma-glutamyl transferase(HS-GGT) fraction and α-fetoprotein mRNA(AFP-mRNA) from peripheral blood mononuclear cells(PBMCs) in diagnosis or monitoring metastasis of HCC. Methods: Total RNA was extracted from hepatomas or PBMCs of patients, synthesized to AFP-cDNA through random primers and reverse transcriptase, amplified by using a nested PCR assay, and confirmed by sequencing. Simultaneity, the HS-GGT activities were quantitatively determined in sera of patients. The comprehensive assessments of two markers were investigated in HCC patients. Results: The amplified fragments of AFP-mRNA were identical to original designed ones(159 bp) and confirmed by sequencing. The lowest sensitivity was 2 ng/L of total RNA. The incidences of AFP-mRNA and HS-GGT were 60.4%(113 of 187) and 84.5%(158 of 187) in the HCC group, and their significantly higher(P < 0.01) than that in any of other groups. The high frequencies of HS-GGT and AFP-mRNA were 78.2% and 56.4% in the HCC patients with AFP < 50 ng/mL. All positive of AFP-mRNA was found in HCC patients with extrahepatic metastasis. No significant correlation was found between tumor size and AFP-mRNA or HS-GGT. The total incidence of combined AFP-mRNA and HS-GGT detection was 93.6 % for HCC diagnosis. Conclusion: HS-GGT and AFP-mRNA are specific markers for HCC diagnosis or differentiation, especially the former for early finding and the later for monitoring metastasis of HCC.

关键词： hepatocellular carcinoma(HCC) AFP-mRNA hepatoma-specific gamma-glutamyl transferase diagnosis metastasis

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肝癌细胞多药耐药逆转技术新进展

中华肝脏病杂志 2014年第9期22卷 715-717页

作者：时运姚敏姚登福南通大学附属医院临床医学研究中心 226001

肝细胞肝癌（HCC）以手术治疗为主,辅之化学治疗、局部治疗等.早期肝癌可选择切除术及肝脏移植,但大部分患者就诊时已失去手术治疗机会[1].尽管新化学治疗药物与方案不断涌现,其效果仍不佳,原因是肝癌存在多药耐药（multidrug resistanc... 详细信息

肝细胞肝癌（HCC）以手术治疗为主,辅之化学治疗、局部治疗等.早期肝癌可选择切除术及肝脏移植,但大部分患者就诊时已失去手术治疗机会[1].尽管新化学治疗药物与方案不断涌现,其效果仍不佳,原因是肝癌存在多药耐药（multidrug resistance,MDR）.药物作用于癌细胞后,其胞内结构、信号通路与调控机制等改变,产生耐药,对药物敏感性降低甚至消失.MDR机制复杂,如细胞表面转运蛋白过表达、凋亡基因失调、酶活性改变等,致化学治疗失败.针对肝癌MDR的逆转方法较多,可降低肝癌MDR,增强化学治疗药物疗效.本文综述了肝癌MDR机制与逆转方法的研究进展.

关键词：癌,肝细胞抗药性,多药逆转方法

Oncofetal glypican-3 is a specific biomarker and valuable gene-therapy for Hepatocellular carcinoma

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Oncofetal glypican-3 is a specific biomarker and valuable ge...