CD147和CK19在肝细胞癌中的表达及临床意义

目的 探讨细胞外基质蛋白诱导因子（CD147）和细胞角蛋白19（CK19）在肝细胞癌（HCC）中的表达及临床意义。方法 采用组织芯片技术和免疫组织化学法检测CD147和CK19在272例HCC组织和81例癌旁组织中的表达情况。结果 CD147在HCC中的阳性表达率为73.53％（200／272）,在癌旁组织中的阳性表达率为13.58％（11／81）,差异有统计学意义（P＜0.05）。CK19在HCC组织中的阳性表达率为14.34％（39／272）,CK19在癌旁组织中无表达。在HCC中,CD147的表达与组织学分级、临床分期、术后无瘤生存时间、肿瘤直径、脉管或门静脉癌栓相关,与性别、年龄、肝硬化、AFP水平、HBV感染、淋巴结转移、病灶数目、肝被膜浸润及卫星灶无关（P＞0.05）;CK19在HCC中的表达与术后无瘤生存时间、组织学分级、肿瘤直径、肝硬化、卫星灶、淋巴结转移及临床分期有关,与性别、年龄、病灶数目、肝被膜浸润、AFP水平、HBV感染及脉管或门静脉癌栓无关（P＞0.05）。在HCC中,CD147 阳性表达者与阴性表达者的中位复发时间分别为13个月和48个月（P＜0.05）,中位生存时间分别为24个月和60个月（P＜0.05）;CK19阳性表达者与阴性表达者的中位复发时间分别为7个月和31个月（P＜0.05）,中位生存时间分别为13个月和42个月（P＜0.05）。 CD147的表达和CK19的表达无明显相关性（r=0.061,P=0.317）。结论 HCC中CD147和CK19的表达与预后密切相关,且两者均可作为HCC预后不良的判断指标。     

食管鳞癌中p53、p57Kip2和CD68的表达及其临床意义（英文）

Objective: The aim of our study was to investigate the expression of p53, p57(Kip2) and CD68 in esophageal squamous cell carcinoma (ESCC) and their correlation with the biological behavior of ESCC. Methods: The protein expressions of p53, p57(Kip2) and CD68 were detected in 51 cases of ESCC with S-P immunohistochemical method. Results: The total positive rate of those proteins was p53 64.71%, CD68 58.82% and p57(Kip2) 45.09% respectively in ESCC. The positive expression rate of p57(Kip2) was significantly lower in the positive p53 of ESCC than in the negative p53 (P < 0.05). The positive expression rate of p57(Kip2) was significantly lower in the positive CD68 of esophageal squamous cell carcinoma than in the negative (P < 0.05). The positive expression of p53 and CD68 were related to differentiate and TNM of ESCC, but p57(Kip2) was not related to TNM (P > 0.05). Conclusion: There are significant negative correlations between p57(Kip2) and p53, CD68 protein expression and related to biological behavior. Multy predictors are better guide to patients than single predictor.     

Downregulation of CD147 expression alters cytoskeleton architecture and inhibits gelatinase production and SAPK pathway in human hepatocellular carcinoma cells

Background

CD147 plays a critical role in the invasive and metastatic activity of hepatocellular carcinoma (HCC) cells by stimulating the surrounding fibroblasts to express matrix metalloproteinases (MMPs). Tumor cells adhesion to extracellular matrix (ECM) proteins is the first step to the tumor metastasis. MMPs degrade the ECM to promote tumor metastasis. The aim of this study is to investigate the effects of small interfering RNA (siRNA) against CD147 (si-CD147) on hepatocellular carcinoma cells' (SMMC-7721) architecture and functions.

Methods

Flow cytometry and western blot assays were employed to detect the transfection efficiency of si-CD147. Confocal microscopy was used to determine the effects of si-CD147 on SMMC-7721 cells' cytoskeleton. Invasion assay, gelatin zymography and cell adhesion assay were employed to investigate the effects of si-CD147 on SMMC-7721 cells' invasion, gelatinase production and cell adhesive abilities. Western blot assay was utilized to detect the effects of si-CD147 on focal adhesion kinase (FAK), vinculiln and mitogen-activated protein kinase (MAPK) expression in SMMC-7721 cells.

Results

Downregulation of CD147 gene induced the alteration of SMMC-7721 cell cytoskeleton including actin, microtubule and vimentin filaments, and inhibited gelatinase production and expression, cells invasion, FAK and vinculin expression. si-CD147 also blocked SMMC-7721 cells adhesion to collagen IV and phosphorylation level of SAPK/JNKs. SAPK/JNKs inhibitor SP600125 inhibited gelatinase production and expression.

Conclusion

CD147 is required for normal tumor cell architecture and cell invasion. Downregulation of CD147 affects HCC cell structure and function. Moreover, the alteration of cell behavior may be related to SAPK/JNK Pathway. siRNA against CD147 may be a possible new approach for HCC gene therapy.     

Overexpression of retinoic acid-induced protein 3 predicts poor prognosis for hepatocellular carcinoma

Aim

To investigate clinical significance of retinoic acid-induced protein 3 (RAI3) in hepatocellular carcinoma (HCC).

Methods

Expression of RAI3 at both mRNA and protein levels in tumor, para-tumor and normal liver tissues was detected in 106 HCC patients by real-time quantitative RT-PCR, Western blot and immunohistochemistry. Then, the correlation of RAI3 expression with clinicopathological characteristics and survivals of HCC patients was analyzed.

Results

Our data first found that RAI3 mRNA and protein expression were both significantly higher in HCC than in para-tumor (both P < 0.001) and normal liver tissues (both P < 0.001). The correlation analysis showed a positive correlation between RAI3 mRNA level and RAI3 protein level in HCC tissues (r = 0.8, P < 0.001). Immunohistochemistry data also revealed that overexpression of RAI3 was present in 73.6 % (78/106) of HCC tissues. In addition, high RAI3 protein expression was correlated with advanced TNM stage (P = 0.001), high serum AFP (P = 0.008), vascular invasion (P = 0.01) and tumor recurrence (P = 0.008). Moreover, HCC patients with overexpression of RAI3 had significantly shorter overall (P = 0.01) and disease-free survival (P = 0.01). Furthermore, multivariate analysis showed that overexpression of RAI3 was an independent prognostic factor for both overall (P = 0.02) and disease-free survival (P = 0.03) in HCC.

Conclusion

Our data for the first time provide a basis for the concept that overexpression of RAI3 may contribute to the malignant progression of HCC and predict poor prognosis for patients with this deadly disease after curative hepatectomy. RAI3 might be an important marker for tumor progression and prognosis, as well as a potential therapeutic target of HCC.     

CK19 and CD10 expression in pancreatic neuroendocrine tumors diagnosed by endoscopic ultrasound‐guided fine‐needle aspiration cytology

BACKGROUND:

CK19 and CD10 are useful markers in the differential diagnosis of pancreatic tumors. The authors evaluated CK19 and CD10 expression in pancreatic neuroendocrine tumors (NETs) obtained by endoscopic ultrasound (EUS)‐guided fine‐needle aspiration (FNA).

METHODS:

Twenty‐eight patients diagnosed with pancreatic NETs based on EUS‐FNA cytology were studied retrospectively (2004‐2007) for immunohistochemical expression of CK19 and CD10. Immunohistochemistry was performed on cell blocks for each case. The pattern of expression for CD10 (cytoplasmic or membranous) and its intensity (0‐2) were noted. The staining of the stromal elements for CD10 was recorded as negative. Cytoplasmic staining in tumor cells and percentage distribution (1+ to 4+) for CK19 were regarded as positive.

RESULTS:

Twenty‐three of 28 (82.14%) NETs showed positive cytoplasmic and/or membranous staining for CD10, and 25 of 28 (89.29%) cases were positive for CK19.

CONCLUSIONS:

The findings demonstrate the high expression of CD10 and CK19 in pancreatic NETs. This indicates that CD10 and CK19 cannot reliably differentiate NETs from other tumors with similar cytomorphologic features (solid pseudopapillary tumors, which frequently stain with CD10, and pancreatic adenocarcinoma, which stains with CK19). Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

The expression and clinical significance of β-catenin and colorectal cancer stem cells marker EpCAMhigh/CD44+ in colorectal cancer

Objective

The aim of the study was to explore the role of Wnt/β-catenin signalling pathway in the maintenance, invasion and metastasis of colorectal cancer stem cells.

Methods

Double immunohistochemical staining was used to detect the expression of EpCAM^high/CD44⁺ which is regarded as the marker of colorectal cancer stem cells in 80 cases of colorectal cancer and their corresponding liver metastases. The SP method of immunohistochemistry was used to detect the expression of the key protein β-catenin in the Wnt pathway in these tissue. The expression and correlation of β-catenin and EpCAM^high/CD44⁺ in colorectal cancer were analyzed and their role on the biological behavior of colorectal cancer was explored.

Results

The abnormal expression of β-catenin was significantly higher in colorectal cancer than in the paraneoplastic normal intestinal mucosa [55% (44/80) vs 10% (2/20), P < 0.05]. The positive expression of EpCAM^high/CD44⁺ was significantly higher in colorectal cancer than in the paraneoplastic normal intestinal mucosa [66.25% (53/80) vs 0% (0/20), P < 0.05]. In the 80 cases of colorectal cancer, the abnormal expression of β-catenin has no correlation with gender (P = 0.079), age (P = 0.416) and the magnitude (P = 0.816) of the tumor (P > 0.05), but it was significantly correlated with degree of differentiation (P = 0.001), depth of invasion (P = 0.001), clinical stage (P = 0.000) and metastasis (P = 0.000). In the colorectal cancer, the expression of EpCAM^high/CD44⁺ cells has no correlation with gender (P = 0.934) and the magnitude (P = 0.160) of the tumor (P > 0.05), but was significantly correlated with age (P = 0.021), degree of differentiation (P = 0.013), depth of invasion (P = 0.000), clinical stage (P = 0.000) and metastasis (P = 0.000). In the corresponding liver metastases, we could also detecte EpCAM^high/CD44⁺ cells. In cases with abnormal expression of β-catenin, the positive expression rate of EpCAM^high/CD44⁺ was significantly higher than those with normal expression of β-catenin (84.1% vs 44.4%), and the difference was statistically significant (P < 0.05).

Conclusion

The abnormal activation of Wnt/β-catenin signalling pathway may prompt the abnormal proliferation of the colorectal cancer stem cells, which leads to the recurrence and metastasis of the cancer.     

肝脏肿瘤组织中HEP、AFP、CK19及CD1O的表达及意义

目的研究HEP、AFP、CK19及CD10在肝脏肿瘤组织中的表达及在鉴别诊断中的应用价值。方法对272例肝细胞肝癌（HCC）、62例胆管细胞癌（BCC）、27例混合性肝癌（CHC）、40例肝脏转移性癌（MC）及10例肝血管平滑肌脂肪瘤（AML）分别进行HEP、AFP、CK19及CD10免疫组织化学标记。结果HEP在HCC中表达率为91．91％，明显高于BCC（9．67％）、MC（12．5％）及AML（0％）（P〈0．01），且HEP表达与HCC的分化相关；AFP在HCC、BCC、CHC、MC及AML中表达率分别为47．79％（130／272），0％（0／62）、48．14％（13／27），10％（4／40），0％（0／10），其在HCC中的表达亦明显高于BCC、MC及AML（P〈0．01）；HCC及BCC中CK19的表达率分别为23．16％（63／272）、100％（62／62），后者明显高于前者（P〈0．01）；而CDl0在HCC中呈现特征性的细胞间小粱状、分支状及逗点状着色，阳性率为50．74％，而在BCC、MC及AML均无此着色。结论联合检测HEP、AFP、CK19及CD10对区别HCC及非HCC具有较高的应用价值。     

MCT4 surpasses the prognostic relevance of the ancillary protein CD147 in clear cell renal cell carcinoma

Cluster of differentiation 147 (CD147/BSG) is a transmembrane glycoprotein mediating oncogenic processes partly through its role as binding partner for monocarboxylate transporter MCT4/SLC16A3. As demonstrated for MCT4, CD147 is proposed to be associated with progression in clear cell renal cell carcinoma (ccRCC). In this study, we evaluated the prognostic relevance of CD147 in comparison to MCT4/SLC16A3 expression and DNA methylation.

Methods

CD147 protein expression was assessed in two independent ccRCC-cohorts (n = 186, n = 59) by immunohistochemical staining of tissue microarrays and subsequent manual as well as automated software-supported scoring (Tissue Studio, Definien sAG). Epigenetic regulation of CD147 was investigated using RNAseq and DNA methylation data of The Cancer Genome Atlas. These results were validated in our cohort. Relevance of prognostic models for cancer-specific survival, comprising CD147 and MCT4 expression or SLC16A3 DNA methylation, was compared using chi-square statistics.

Results

CD147 protein expression generated with Tissue Studio correlated significantly with those from manual scoring (P < 0.0001, r_S = 0.85), indicating feasibility of software-based evaluation exemplarily for the membrane protein CD147 in ccRCC. Association of CD147 expression with patient outcome differed between cohorts. DNA methylation in the CD147/BSG promoter was not associated with expression. Comparison of prognostic relevance of CD147/BSG and MCT4/SLC16A3, showed higher significance for MCT4 expression and superior prognostic power for DNA methylation at specific CpG-sites in the SLC16A3 promoter (e.g. CD147 protein: P = 0.7780, Harrell''s c-index = 53.7% vs. DNA methylation: P = 0.0076, Harrell''s c-index = 80.0%).

Conclusions

Prognostic significance of CD147 protein expression could not surpass that of MCT4, especially of SLC16A3 DNA methylation, corroborating the role of MCT4 as prognostic biomarker for ccRCC.