GENERATION AND CHARACTERIZATION OF TWO MONOCLONAL ANTIBODIES AGAINST CYTOKERATINS

Thirty-five hybridoma cell lines against cytoke-ratins, isolated from Hela cells and human cellus respectively, were generated by fusion of immunized spleen cells of BALB/C mice with P3×63-Ag8.653, a mouse myeloma cell line. Two of them (HI and C53) were characterized by indirect immu-nofluorescence, ABC immunostaining and immuno-blotting. The results of immunofluorescence and ABC immunostaining suggested that both monoclonal antibodies were specific for keratin-type intermediate filaments. However, the two monoclonal antibodies showed different specificities in normal tissues and neoplasms as observed on both frozen and deparaf-finized sections. In normal tissues, H1 stained transitional epithelium and all types of simple epithelium except endothelium and mesothelium but did not stain stratified squamous epithelium. In contrast, C35 recognized only stratified squamous epithelium, but failing to react with simple epithelium. Both monoclonal antibodies did not react with nonepithelia cells and tissues. In neoplasms, H1 stained varieties of adenocaroinomas and C35 recognized merely squamous cell carcinomas. Therefore, all the epithelial tissues can nearly be recognized by combination of the two monoclonal antibodies. All the results indicated that H1 and C35 can be used in cell biology and histology studies, and can be used in differential diagnosis of adenocarcinoma and squamous cell carcinoma in pathology.

HISTOLOGICAL RECOGNITION SPECTRUMS OF 4 MONOCLONAL ANTIBODIES AGAINST CYTOKERATINS

The histological specificities of 4 monoclonal antibodies against cytokeratins (HK2, HK5, K174 and K27) were investigated in various kinds of human and rat tissues with ABC immunohistochemical technique. K174 was shown to have the same recognition spectrum with a polyclonal antibody (RAK1) to epidermal keratin. HK2 and HK5 were similar except the difference in reaction with stratified squqmous epithelium. K27 could only label the suprabasal layers of the squamous epithelium, and could be used as a marker of cornified or cornifying epithelium. This study provided a sound basis for the use of this group of antibodies in the subtyping of different epithelial tissues and the tumors originating from them.

THE EXPRESSION OF CYTOKERATINS IN HUMAN HEPATOCELLULAR AND CHOLANGIOCELLULAR CARCINOMAS

In order to determine the usefulness value of the antibodies to cytokeratins(CK) of'bile duct type'in the differential diagnosis between hepatocellular carcinoma(HCC) and cholangiocellular carcinoma(CC),we have made an immunocytochemical investigation,using the antibodies specifically recognizing CK19 and CK18,seperately,in liver,and laminin(LN) antibody.All the CC examined(10 cases) were found CK19-positive;interestingly,CK19-positive cancer cells were also observed in 38% of HCCs(14/37).Therefore,CK19 was not a reliable marker in differentiating HCC from CC,in our consideration.The CK19 expression in HCC was showed to be irrelevant to their differentiation degres,but related to the histologic subtypes which indicated the directions of their differentiation.CK19 expression was observed in all the HCC cell nests with glandular differentiation,and an untontinuous LN-Positive basement membrane-like structure was immunolocalized around these cells.Which indicated that the glandular differentiation and CK19 expression in HCC were also related to the LN deposition,as in fetal liver and some chronic liver disorders.

桥本甲状腺炎不典型细胞群Galectin-3、CK19、CK7的表达及意义

目的探讨桥本甲状腺炎(Hashimoto′s thyroiditis,HT)中不典型细胞群与甲状腺乳头状癌(papillary thyroid carcinoma,PTC)的关系及其临床病理意义。方法从存档病例中选取45例PTC和40例HT,以40例腺瘤周围甲状腺组织为对照,进行常规形态学观察和Gal-3、CK19和CK7、MaxVision法免疫组化染色,以χ2检验分析表达结果。结果(1)HT增生的淋巴滤泡周围可见到PTC-核样改变的不典型细胞群;(2)Gal-3在PTC,HT不典型细胞群及对照组表达率分别为89%,10%,0,差异有统计学意义(P<0.05),前组高于后两组,后两组间差异无统计学意义(P>0.05);(3)CK19和CK7在PTC,HT不典型细胞群及对照组的阳性表达率分别为100%,95%,10%和95%,92.5%,12.5%,两种抗体的阳性率和表达部位均较接近,前两组均高于对照组(P<0.05),PTC和HT两组间差异无统计学意义(P>0.05)。结论Gal-3在HT不典型细胞群和PTC组表达的差异性可协助诊断HT是否合并PTC;CK19和CK7在HT不典型细胞群中与PTC组有类似的表达模式,提示HT不典型细胞群可能是PTC的前驱病变,应密切随访,谨防癌变。

Cytokeratin 19、HBME-1和Galectin-3在甲状腺乳头状癌诊断中的应用

目的探讨Cytokeratin 19(CK19)、HBME-1和Galectin-3在甲状腺乳头状癌和乳头状增生性甲状腺疾病鉴别诊断中的作用。方法收集289例甲状腺切除标本,包括144例经典型乳头状癌(CPC)、21例滤泡型乳头状癌(FVPC)、19例乳头状微小癌(PM)、46例结节性甲状腺肿伴乳头状增生(NGWPH)和59例甲状腺腺瘤伴乳头状增生(FAWPH),采用EnVision二步法检测CK19、HBME-1和Galectin-3的表达。结果 CK19、HBME-1和Galectin-3在CPC中的阳性表达率分别为100%、93.1%和89.6%,在FVPC中为100%、76.2%和76.2%,在PM中为100%、73.7%和78.9%,在NGWPH中为5.1%、2.2%和4.3%,在FAWPH中为4.3%、1.7%和5.1%。CK19、HBME-1和Galectin-3的阳性表达率在乳头状癌与乳头状增生性甲状腺疾病之间比较,差异均有统计学意义(P<0.01)。结论在甲状腺乳头状癌鉴别诊断中,CK19的敏感性较高,HBME-1和Galectin-3特异性较强;三种抗体联合应用,有助于鉴别甲状腺乳头状癌与乳头状增生。

Hep Par 1、CD34及Cytokeratin在血清AFP阴性肝细胞癌诊断中的应用

目的:探讨HepPar1、CD34及Cytokeratin在血清[AFP(-)HCC]诊断及其在ICC、MAC鉴别诊断中的意义.方法:选取南通市肿瘤医院1989-2007年手术切除的70例[AFP(-)HCC]、6例ICC及24例MAC的石蜡标本,免疫组织化学染色法检测HepPar1、CD34及Cytokeratin在血清[AFP(-)HCC]中的表达.结果:HepPar1、CD34在血清AFP(-)HCC、ICC及MAC中的表达差异性均具有显著统计学意义(HepPar1:χ2=50.7937、9.5745及37.4532;CD34:χ2=67.0330、9.9836及49.3927,均P<0.01).在分化不良组与分化较好组血清AFP(-)HCC中表达差异性的比较中,HepPar1与CD34、CK20、CK19的表达均有统计学意义(P<0.01或0.05).联合应用HepPar1、CD34对AFP(-)HCC与ICC、MAC的鉴别诊断评价准确度、灵敏度及特异度分别为:90.7%、89.8%及93.3%.结论:联合应用HepPar1、CD34及Cytokeratin可提高血清AFP(-)HCC的诊断准确率及其与ICC、MAC的鉴别诊断准确率,为临床治疗方案的选择与预测评估提供了病理依据.

Cytokeratms and carcinoembryonic antigen in diagnosis,staging and prognosis of colorectal adenocarcinoma

AIM: To evaluate the serum levels of cytokeratins and carcinoembryonic antigen (CEA) in diagnosis, staging and prognosis of patients with colorectal adenocarcinoma.METHODS: The sample consisted of 169 patients. One hundred blood donors formed the control group. Radical surgery was performed on 120 patients, with an average follow-up duration of 22.3 mo. Relapses occurred in 23individuals after an average of 18.09 mo. CEA was assayed via the Delfia(R) method with a limit of 5 ng/mL. Cytokeratins were assayed via the LIA-mat(R) TPA-M Prolifigen(R) method with a limit of 72 U/L.RESULTS: In the diagnosis of patients with colorectal adenocarcinoma, CEA showed a sensitivity of 56%, a specificity of 95%, a positive predictive value of 94%, a negative predictive value of 50% and an accuracy of 76.8%.TPA-M had a sensitivity of 70%, a specificity of 96%, a positive predictive value of 97%, a negative predictive value of 66% and an accuracy of 93.6%. The elevation of one of the markers was shown to have a sensitivity of 76.9%, a specificity of 91%, a positive predictive value of 93.5%, a negative predictive value of 70% and an accuracy of 83.6%.There was no variation in the levels of the markers according to the degree of cell differentiation while there was an elevation in their concentrations in accordance with the increase in neoplastic dissemination. There was a statistically significant difference between the patients with stage Ⅳ lesions and those with stages Ⅰ, Ⅱ and Ⅲ tumors.With regard to CEA, the averagelevel was 14.2 ng/mL in patients with stage Ⅰ lesions, 8.5 ng/mL in patients with stage Ⅱ lesions, 8.0 ng/mL in patients with stage Ⅲ lesions and 87.7 ng/mL in patients with stage Ⅳ lesions. In relation to TPA-M, the levels were 153.1 U/L in patients with stage Ⅰtumors, 106.5 U/L in patients with stage Ⅱ tumors, 136.3 U/L in patients with stage Ⅲ tumors and 464.3 U/L in patients with stage Ⅳ tumors. There was a statistical difference in patients with a high CEA level in relation to a shorter survival(P＜0.05). However, there was no correlation between patients with high TPA-M levels and prognostic indices of patients undergoing radical surgery.CONCLUSION: Cytokeratins demonstrate a greater sensitivity than CEA in the diagnosis of colorectal adenocarcinoma.There is an increase in the sensitivity of the markers with tumor dissemination. Cytokeratins cannot identify the worse prognosis in patients undergoing radical surgery.Cytokeratins constitute an advance in the direction of a perfect tumor marker in the treatment of patients with colorectal cancer.

P-gp170、cytokeratin及nm23在人涎腺粘液表皮样癌MEC-1/5Fu耐药细胞中的表达

目的 :探讨人涎腺粘液表皮样癌耐药细胞株MEC 1/ 5Fu细胞的耐药机制。方法 :应用免疫组织化学SP方法观察该细胞及其亲本细胞中P 糖蛋白 (P gp170 )、cytokeratin(CK)及nm 2 3的表达。 结果 :P gp170在MCE 1/ 5Fu细胞中的阳性表达率为 95 .1% ,明显高于亲本细胞的阳性表达率 12 .3 % (P <0 .0 5 ) ,而CK与nm2 3在MEC 1/ 5Fu细胞与亲本细胞中的表达率分别达 90 %以上 ,无显著差异 (P >0 .0 5 )。结论 :MEC 1/5Fu细胞具有多药耐药 (MDR)表型 ,其耐药性的产生主要是由P

肝硬化组织中卵圆细胞的光镜、电镜与免疫电镜观察

目的 :探讨人肝硬化肝组织是否存在卵圆细胞。方法 :对 30例人肝细胞肝癌手术标本的癌旁肝硬化组织行常规组织学观察 ,并用胆管上皮分化标记CK7和肝细胞分化标记白蛋白对以上组织作免疫组化染色 ,同时对其中 10例作了超微结构观察 ,5例作免疫电镜标记和观察。另外对 5例正常肝脏和 5例肝炎后肝硬化组织同时作了光镜、免疫组化和电镜观察 ,对 2例肝炎后肝硬化作了免疫电镜标记和观察。结果 :癌旁肝硬化组织的 30例光镜和 10例电镜观察以及 5例肝炎肝硬化光、电镜观察 ,均可在再生的肝细胞结节边缘见到散在的小细胞和增生的小胆管样结构。这些小细胞和增生的小胆管内少数小细胞 ,为卵圆形 ,体积较小 ,核大 ,胞质少 ,胞质内含较多的游离核糖体 ,仅含少量粗面内质网和线粒体 ,胞质内可见张力微丝结构 ,这类细胞与邻近细胞间均有细胞间连接。免疫电镜示 ,卵圆细胞均表达CK7和白蛋白 ,但有些细胞内表达CK7多些 ,有些细胞则表达白蛋白多些。在正常肝组织内未见到类似细胞。结论 :与动物实验性肝癌模型肝组织一样 ,人类肝硬化肝组织中也存在同样细胞形态和免疫表型特点的卵圆细胞。

大肠癌患者外周血CK19和MUC-1 mRNA水平及其临床意义

目的探讨大肠癌(CRC)患者外周血中角蛋白(CK)19、黏蛋白(MUC)-1 mRNA水平并分析两者与临床病理特征的关系。方法采用巢式逆转录聚合酶链式反应(RT-PCR)检测20例健康体检者(健康组)、20例大肠良性腺瘤患者(良性疾病组)及90例CRC患者(CRC组)的外周血CK19和MUC-1 mRNA水平,并分析两者与CRC临床病理参数(Dukes分期、分化程度、远处转移及组织学类型)的关系。结果 CRC组CK19 mRNA阳性率为58.9%(53/90),均高于健康组的0和良性疾病组的5.0%(1/20),差异有统计学意义(P<0.05);CRC组MUC-1 mRNA阳性率为52.2%(47/90),与健康组的60.0%(12/20)和良性疾病组的50.0%(10/20)比较,差异无统计学意义(P>0.05);健康组和良性疾病组CK19 mRNA、MUC-1mRNA阳性率的差异无统计学意义(P>0.05)。CRC中CK19 mRNA和MUC-1 mRNA阳性率均与Dukes分期、分化程度及远处转移有关(P<0.05),但与组织学类型无关(P>0.05)。结论 CRC患者外周血中CK19 mRNA阳性率明显升高,且外周血中CK19mRNA、MUC-1 mRNA与Dukes分期、分化程度及远处转移有关,提示两者可能在CRC发生发展中均有重要意义。

不同分离纯化法建立人早孕滋养细胞模型的效果比较

目的对4种建立滋养细胞的方法进行比较,以期寻求一种简便高效的人早孕滋养细胞的体外培养方法,为相关研究提供基础。方法早孕绒毛通过胰酶联合胶原酶消化分离后,分别采用直接接种、差速贴壁联合差速消化法、人外周血淋巴细胞分离液分离纯化法、完整绒毛消化联合简化的percoll密度梯度分离法4种方法纯化培养,倒置显微镜观察细胞形态,应用HE染色、免疫组化和免疫细胞化学法检测细胞纯度。结果直接接种、差速贴壁联合差速消化法、人外周血淋巴细胞分离液分离纯化法得到的细胞细胞角蛋白7(CK-7)表达阳性率不足60%;简化的percoll密度梯度分离法得到的细胞CK-7表达阳性率达90%以上。结论完整绒毛消化联合简化的percoll密度梯度分离纯化可以得到大量较高纯度的滋养细胞以供后期实验。

血清AFP阴性肝细胞肝癌免疫组化诊断谱研究

目的探讨Hep Par1等抗体在诊断血清AFP阴性肝细胞肝癌(HCC)及鉴别肝内胆管细胞癌(ICC)、肝转移性腺癌(MAC)中的意义。方法选取手术切除100例血清AFP阴性HCC、30例ICC及36例MAC(胃腺癌30例、肺腺癌6例)石蜡标本,根据Hep Par1等抗体的染色、表达率、金标准诊断、综合性能计分(CCS)筛选免疫组化诊断谱。结果血清AFP阴性HCC中,Hep Par1胞浆染色(CCS≥8);CD34血管均匀染色(CCS=9);CD10(CCS=8)及CEA毛细胆管染色。ICC、MAC中,CK19(ICC中CCS=7)、CK7(MAC中CCS=7)胞浆染色,CEA膜浆染色。结论诊断血清AFP阴性HCC联用CD34与Hep Par1(或CD10、CEA);鉴别诊断ICC、MAC分别加用CK19、CK7。

Modeling and validating three dimensional human normal cervix and cervical cancer tissues in vitro

The use of three dimensional in vitro systems in cancer research is a promising path for developing effective anticancer therapies.The aim of this study was to engineer a functional 3-D in vitro model of normal and cancerous cervical tissue.Normal epithelial and immortalized cervical epithelial carcinoma cell lines were used to construct 3-D artificial normal cervical and cervical cancerous tissues.De-epidermised dermis（DED） was used as a scaffold for both models.Morphological analyses were conducted by using hematoxylin and eosin staining and characteristics of the models were studied by analyzing the expression of different structural cytokeratins and differential protein marker MAX dimerisation protein 1（Mad1） using immunohistochemical technique.Haematoxylin and eosin staining results showed that normal cervical tissue had multi epithelial layers while cancerous cervical tissue showed dysplastic changes.Immunohistochemistry staining revealed that for normal cervix model cytokeratin 10 was expressed in the upper stratified layer of the epithelium while cytokeratin 5 was expressed mainly in the middle and basal layer.Cytokeratin 19 was weakly expressed in a few basal cells.Cervical cancer model showed cytokeratin 19 expression in different epithelial layers and weak or no expression for cytokeratin 5 and cytokeratin 10.Madl expression was detected in some suprabasal cells.The 3-D in vitro models showed stratified epithelial layers and expressed the same types and patterns of differentiation marker proteins as seen in corresponding in vivo tissue in either normal cervical or cervical cancerous tissue.These findings imply that they can serve as functional normal and cervical cancer models.

胃黏膜内癌淋巴结微转移的临床意义

目的：研究胃黏膜内癌淋巴结微转移的发生率、病理学特征及其临床意义.方法：胃黏膜内癌患者84例手术切除淋巴结2526枚进行连续超薄切片,分别进行HE染色及抗Cytokeratin(CK；CAM5.2)免疫组化研究,并与临床病理资料进行对比分析.结果：在84例中16例患者具有淋巴结受累(19%)；2526枚淋巴结中45枚淋巴结受累(1.8%；45/2526),显著高于HE染色的1.2% (1/84；P＜0.05),淋巴结微转移率为18% (15/84).尽管没有显著统计学差异,微转移在大于1.0cm的肿瘤(15/72；21%)较小于或等于1.0cm的肿瘤(1/12；8%)更为多见(P=0.307).大于2.0cm的肿瘤,淋巴结的微转移均为淋巴结内多发散在或聚集状态的肿瘤细胞.结论：胃黏膜内癌具有较高的淋巴结微转移率,内镜下黏膜切除术不宜用于直径大于1.0cm的胃黏膜内癌.

High yield reproducible rat model recapitulating human Barrett's carcinogenesis

AIM To efficiently replicate the biology and pathogenesis of human esophageal adenocarcinoma(EAC) using the modified Levrat model of end-to-side esophagojejunostomy. METHODS End-to-side esophagojejunostomy was performed on rats to induce gastroduodenoesophageal reflux to develop EAC. Animals were randomly selected and serially euthanized at 10(n = 6),17(n = 8),24(n = 9),31(n = 6),38(n = 6),and 40(n = 6) wk postoperatively. The esophagi were harvested for downstream histopathology and gene expression. Histological evaluation wascompleted to determine respective rates of carcinogenic development. Quantitative reverse transcriptionpolymerase chain reaction was performed to determine gene expression levels of MUC2,CK19,and CK20,and results were compared to determine significant differences throughout disease progression stages.RESULTS The overall study mortality was 15%. Causes of mortality included anastomotic leak,gastrointestinal hemorrhage,stomach ulcer perforation,respiratory infection secondary to aspiration,and obstruction due to tumor or late anastomotic stricture. 10 wk following surgery,100% of animals presented with esophagitis. Barrett's esophagus(BE) was first observed at 10 wk,and was present in 100% of animals by 17 wk. Dysplasia was confirmed in 87.5% of animals at 17 wk,and increased to 100% by 31 wk. EAC was first observed in 44.4% of animals at 24 wk and increased to 100% by 40 wk. In addition,two animals at 38-40 wk post-surgery had confirmed macro-metastases in the lung/liver and small intestine,respectively. MUC2 gene expression was progressively down-regulated from BE to dysplasia to EAC. Both CK19 and CK20 gene expression significantly increased in a stepwise manner from esophagitis to EAC. CONCLUSION Esophagojejunostomy was successfully replicated in rats with low mortality and a high tumor burden,which may facilitate broader adoption to study EAC development,progression,and therapeutics.

造釉细胞瘤45例临床病理分析

造釉细胞瘤(Ameloblastoma)是一种常见的牙源性肿瘤,其发病率几乎等于除牙瘤外所有其它牙源性肿瘤的发病率的总和。该瘤具有多变的组织结构特点。1974～1992年间我院收治45例报告如下。

ki67、Cytokeratin19在口腔粘膜扁平苔藓及口腔鳞癌上皮组织中表达的研究

目的研究口腔粘膜扁平苔藓及口腔鳞癌患者口腔粘膜上皮ki67,Cytokeratin19的表达,探讨OLP发病的可能机理及是否为癌前病变。方法用免疫组化SABC法检测20例口腔粘膜扁平苔藓患者口腔粘膜组织及30例OSCC口腔粘膜组织中ki67,Cytokeratin19的表达并行细胞计数,其中对Cytokeratin19的表达行半定量统计分析。结果(1)OLP与正常组中ki67中阳性表达率有显著差异,P<0.05;(2)Cytokeratin19行半定量分析发现OLP组与OSCC组有显著差异,P<0.05。结论(1)ki67,Cytokeratin19的异常表达可能是OLP,OSCC的发病原因。(2)Cytokertin19、ki67的高表达均提示OLP可能是一种癌前病变。(3)Cytokertin19、ki67可作为口腔鳞癌检测的指标。

妊娠滋养细胞疾病中EMT相关蛋白CK19和N-cad表达的研究

目的:探讨上皮细胞向间质转化(EMT)相关蛋白细胞角蛋白19(CK19)和神经钙黏蛋白(N-cad)在妊娠滋养细胞疾病发生、发展中的作用。方法:采用免疫组化SP法检测了41例正常早孕绒毛(NP)、31例葡萄胎(HM)、32例妊娠滋养细胞肿瘤(GTN)[26例侵蚀性葡萄胎(IM)+6例绒癌(CCA)]中CK19和N-cad的定位及表达情况。结果:NP组中CK19和N-cad蛋白相对表达均显著高于HM、GTN组(P<0.05),HM组显著高于GTN组(P<0.05)。CK19与N-cad的表达无相关性(r=0.202,P=0.268)。结论:伴随着滋养细胞恶性程度的升高,CK19和N-cad表达逐渐降低,提示EMT可能与滋养细胞的恶性转化有关。

Prognostic significance of preoperative and postoperative CK19 and CEA m RNA levels in peripheral blood of patients with gastric cardia cancer

AIM To evaluate the clinical and prognostic significance of preoperative and postoperative cytokeratin 19(CK19) and carcinoembryonic antigen(CEA) m RNA levels in peripheral blood of patients with gastric cardia cancer(GCC).METHODS We detected the preoperative and postoperative mR NA levels of CK19 and CEA in peripheral blood of 129 GCC patients by using reverse transcription-polymerase chain reaction and evaluated their clinical and prognostic significance by univariate Kaplan-Meier survival analysis and multivariate Cox proportional hazard analysis. A new prognostic model which stratified patients into three different risk groups was established based on the independent prognostic factors.RESULTS Elevated preoperative and postoperative CK19 and CEA mR NA levels in peripheral blood of GCC patients were associated with lymph node metastasis. Univariate analysis showed that tumor size, histological grade, depth of tumor invasion, lymph node metastasis, preoperative CK19 m RNA, and preoperative and postoperative CEA m RNA levels were correlated with the prognosis of GCC patients. The multivariate analysis showed that lymph node status(P = 0.018), preoperative CK19(P = 0.035) and CEA(P = 0.011) m RNA levels were independent prognostic factors for overall survival(OS). The 5-year OS rates for the low-, intermediate-, and high-risk groups were 48.3%, 22.6%, and 4.6%, respectively(P < 0.001).CONCLUSION Elevated preoperative CK19 and CEA mR NA levels may be regarded as promising biomarkers for predicting lymph node metastasis and poor prognosis in patients with GCC. This new prognostic model may help us identify the subpopulations of GCC patients with the highest risk.

Prognosis of hepatocellular carcinoma expressing cytokeratin 19:Comparison with other liver cancers

AIM:To investigate whether expressing biliary phenotype predicted poor outcome after the surgical treatment in primary liver cancers. METHODS:Out of 204 patients that underwent liver resection due to hepatocellular carcinoma (HCC), liver specimens of 70 patients with HCC were evaluated for biliary components by cytokeratin (CK) 19 immunostain (CK19 - HCC and CK19 + HCC). CK19 positivity was defined as membranous and/or cytoplasmic expression in ≥ 5% of tumor cells with moderate or strong intensity. Patients with other primary liver cancers, such as com- bined HCC and cholangiocarcinoma (cHCC-CC), intrahe- patic cholangiocarcinoma (ICC) who received curative liver resection, were also included in the study. Clinical characteristics of CK19-HCC and CK19 + HCC patients, including survival outcome after curative liver resection, were compared with that of cHCC-CC and ICC patients. RESULTS: The overall survival (OS) rate of CK19 - HCC(n = 49) after the curative surgical treatment was 90.7%, and 80.4% at 1 and 5 years after the resection. OS rate of CK19 + HCC (n = 21) was 74.3%, 28.9% and OS rate of cHCC-CC (n = 22) was 66.7%, 32.2% at 1 and 5 years after the surgery. For ICC (n = 19), 1 and 5-year-OS rate was 50.2% and 14.3% after the cura-tive resection. The OS rates of CK19 + HCC and cHCC-CC were significantly lower than that of CK19-HCC, but higher than the OS rate of ICC (P = 0.000). There was no statistically significant difference in OS rate between CK19 + HCC and cHCC-CC. The disease free survival (DFS) rate of CK19-HCC was 72.0% and 54.5% at 1 and 3 years after the surgical treatment. DFS rate of CK19 + HCC was 53.3%, 34.3% and DFS rate of cHCC- CC was 51.5%, 39.2% at 1 and 3 years after the resection. For ICC, 1 and 3-year-DFS rate was 28.0% and 14.0% after the curative resection. DFS rate of CK19-HCC was significantly higher than that of ICC (P = 0.017), but marginally higher than DFS rate of either CK19 + HCC or cHCC-CC (P = 0.097, P = 0.089, respec-tively). Predictors of outcome after the surgery of primary liver cancer were pathology of the resected mass, existence of microvascular invasion and accompanying satellite nodule. CONCLUSION: Primary liver cancers with biliary components tended to show poorer surgical outcome. This suggested that immuno-phenotype of liver cancers was as important as their morphological classification.