癌相关分子CD147与肝癌靶向治疗

CD147是一种在肝癌细胞膜表面高表达的跨膜糖蛋白,能够调节肝癌细胞生长、诱导基质金属蛋白酶（matrix metalloprotei-nase,MMP）分泌、促进肝癌细胞侵袭和转移,并且参与肝癌血管生长和耐药性形成。以CD147为靶点的单克隆抗体治疗肝癌具有显著疗效。本文就肝癌细胞CD147的分子特点、功能与机理以及针对CD147的靶向治疗等方面研究进展作较全面的综述。     

人CD147的克隆表达及其对成纤维细胞基质金属蛋白酶表达的影响

CD147可以促进基质金属蛋白酶(MMPs)的表达,与肿瘤的生长和浸润有关。为了研究CD147在大肠癌中的作用,利用RT-PCR从一健康人克隆了cd147基因,测序发现该基因存在两个碱基突变,其中C634T造成了CD147跨膜区212位氨基酸由L突变为F。分别构建CD147的原核(pGEX-5x-147)和真核(pEGFP-147)表达系统,在宿主菌BL21和CCL229细胞中均获得了稳定表达。Western印迹显示原核表达产物比真核CD147分子量小,说明原核CD147缺乏糖基化。荧光显微镜显示真核CD147表达定位于CCL229细胞膜,表明突变L212F不影响CD147的膜定位。用明胶电泳检测表达的CD147对MMPs表达的影响,结果显示原核产物不能诱导MMPs表达上调,而真核产物能够明显诱导MMPs表达上调,说明糖基化对于CD147活性是必需的,真核系统能够表达具有生物功能的CD147,并且突变L212F不会影响蛋白质的活性。     

Role of CD10 immunochemistry in differentiating hepatocellular carcinoma from metastatic carcinoma of the liver

Background:  Differentiation of hepatocellular carcinoma (HCC) from metastatic carcinoma in liver may be difficult on fine needle aspiration cytology (FNAC), especially when both appear as moderate to poorly differentiated tumours. A panel of immunocytochemical stains is frequently used in case of diagnostic difficulty. Recently, CD10 immunostain with a canalicular staining pattern has been shown to be a specific marker for hepatocytic differentiation.
Objective:  The present study was designed to assess the value of CD10 immunostain in distinguishing HCC from metastatic carcinoma in material obtained by FNAC of liver masses.
Materials and methods:  Formalin-fixed, paraffin-embedded cell blocks of 22 cases (7 cases of HCC and 15 cases of metastatic carcinoma), direct acetone-fixed smears and destained smears of 28 cases (18 cases of HCC and 10 cases of metastatic carcinoma) prepared from FNAC of the liver were immunostained using monoclonal antibody against CD10.
Results:  Seventeen (68%) of twenty-five cases of HCC were positive for CD10 with a canalicular staining pattern. Among them 7 (70%) of 10 cases were well-differentiated HCC and 10 (66%) of 15 cases were moderate to poorly differentiated HCC. Of 25 cases of metastatic carcinoma, four (16%) were positive for CD10 with a cytoplasmic (three cases) and membranous staining (one case) pattern.
Conclusion:  CD10 immunostaining is useful in discriminating HCC and metastatic carcinoma of the liver and is easily applied on cell blocks as well as FNAC smears.     

CD147 promotes progression of head and neck squamous cell carcinoma via NF‐kappa B signaling

CD147/basigin (BSG) is highly upregulated in many types of cancer, our previous study has found that CD147/BSG is highly expressed in head and neck squamous cell carcinoma (HNSCC) stem cells, but its role in HNSCC and the underlying mechanism is still unknown. In this study, we investigated the role of CD147 in the progression of HNSCC. Real‐time PCR, western blot and immunohistochemistry were used to detect the expression of CD147 in total 189 HNSCC tissues in compared with normal tissues. In addition, we used proliferation, colony formation, cell cycle and apoptosis, migration and invasion as well as wound‐healing assay to determine the biological roles of CD147 in HNSCC. Then, a xenograft model was performed to evaluate tumor‐promoting and metastasis‐promoting role of CD147 in HNSCC. The results showed that upregulated CD147 expression was associated with aggressive clinicopathologic features in HNSCC. In addition, CD147 promoted proliferation, migration and reduced the apoptosis phenotype of HNSCC cells in vitro as well as tumor initiation and progression in vivo. Furthermore, we demonstrated that CD147 promoted HNSCC progression through nuclear factor kappa B signaling. Therefore, we concluded that CD147 promoted tumor progression in HNSCC and might be a potential prognostic and treatment biomarker for HNSCC.     

TMAO通过CD147/MMPs通路诱导巨噬细胞活化

肠道菌群代谢产物氧化三甲胺(trimethylamine N-oxide,TMAO)可通过多种途径促进动脉粥样硬化(atherosclerosis,AS)的进展,现研究发现,在临床上其与斑块稳定性存在密切联系,但其分子机制目前尚不明确.金属蛋白酶诱导因子(extracellular matrix metalloprot...     

Novel Functions of CD147 in the Mitochondria Exacerbates Melanoma Metastasis

Melanoma is an aggressive form of skin cancer characterized by rapid invasion and metastasis. CD147 is known to be functioning in cell invasion. In this study, we showed that CD147 was translocated from the cell membrane to the mitochondria in advanced melanoma. Melanoma patients with CD147 localized to the mitochondria confer a worse prognosis. The mitochondrial CD147 levels are correlated with the invasion potential of various melanoma cell lines as well as mitochondrial energy metabolism. Depletion of CD147 decreased the activity of mitochondrial complex V. STRING analysis for protein-protein interaction networks (PPIN) in CD147-depleted melanoma cells showed that mitochondrial proteins HSP60 and ATP5B, a subunit of mitochondrial complex V, were node proteins. HSP60 upregulation was correlated with a worse prognosis of melanoma patients. Co-immunoprecipitation (Co-IP) assay indicates that CD147 interacts with HSP60. These data suggested that mitochondrial CD147 may prompt HSP60 to activate ATP5B, thereby promoting the mitochondrial aerobic oxidation and the invasive abilities of melanoma cells. Correlation analysis of the data acquired from patients was helpful to draw a 5-year survival curve for patients who screened positive and negative for mitochondrial CD147. This study unravels the function of CD147 in tumor invasion and highlights it as a potential tumor therapeutic target.     

EMMPRIN (CD147) expression and differentiation of papillary thyroid carcinoma: implications for immunocytochemistry in FNA cytology

Y. Aratake, K. Marutsuka, K. Kiyoyama, T. Kuribayashi, T. Miyamoto, K. Yakushiji, S. Ohno, Y. Miyake, T. Sakaguchi, T. K. Kobayashi, A. Okayama, K. Tamura and E. Ohno
EMMPRIN (CD147) expression and differentiation of papillary thyroid carcinoma: implications for immunocytochemistry in FNA cytology Objective: Extracellular matrix metalloproteinase inducer (EMMPRIN) and its induced matrix metalloproteinases (MMPs) play a crucial role in tumour progression, invasion and metastasis. EMMPRIN expression has been demonstrated in several tumours, but its expression profile in thyroid cancer remains unclear. Methods: We evaluated the expression profile of EMMPRIN at various stages of differentiation of thyroid carcinoma, including 20 cases of well‐differentiated papillary carcinoma (WDPC), 15 cases of papillary carcinoma with a poorly differentiated carcinoma component (PC/PDC) and four cases with an undifferentiated carcinoma (UDC) component, using paraffin‐embedded sections for immunohistochemical stains. Also, we used 32 fine needle aspiration cytology and imprint smears from the same cases for immunocytochemical stains. The staining results were evaluated with a scoring system. Results: Immunohistochemical staining showed that EMMPRIN expression was absent or weak in almost all WDPC specimens, whereas it was moderate or strong in PDC and UDC components. In tumours that showed a gradual morphological transformation from WDPC to PDC components, the expression of EMMPRIN was progressively stronger from the areas of WDPC to those of PDC. WDPC, PC/PDC and UDC had expression scores of 4.9, 45.0 and 245.7, respectively. Results of immunocytochemical staining showed almost the same staining profile as those of immunohistochemical staining. The cytological atypia of EMMPRIN‐positive cells was greater than that of negative cells. Conclusion: These results indicated that EMMPRIN expression correlates significantly with the degree of dedifferentiation of thyroid carcinoma. This study demonstrates the feasibility of expression of EMMPRIN using fine needle aspiration samples. Therefore, immunocytochemical analysis of EMMPRIN may be a novel aid to evaluate the differentiation of thyroid carcinoma.     

CD63 negatively regulates hepatocellular carcinoma development through suppression of inflammatory cytokine-induced STAT3 activation

Tetraspanin CD63 has been widely implicated in tumour progression of human malignancies. However, its role in the tumorigenesis and metastasis of hepatocellular carcinoma (HCC) remains unclear yet. In the present study, we aimed to investigate the specific function and underlying mechanisms of CD63 in HCC progression. CD63 expression in HCC tissues was detected using immunohistochemistry and quantitative real-time PCR analyses; effects of CD63 on HCC cell proliferation and migration were investigated by CCK-8 assay, colony formation assay, transwell assay and a xenograft model of nude mice. RNA-sequencing, bioinformatics analysis, dual-luciferase reporter assay and Western blot analysis were performed to explore the underlying molecular mechanisms. Results of our experiments showed that CD63 expression was frequently reduced in HCC tissues compared with adjacent normal tissues, and decreased CD63 expression was significantly associated with larger tumour size, distant site metastasis and higher tumour stages of HCC. Overexpression of CD63 inhibited HCC cell proliferation and migration, whereas knockdown of CD63 promoted these phenotypes. IL-6, IL-27 and STAT3 activity was regulated by CD63, and blockade of STAT3 activation impaired the promotive effects of CD63 knockdown on HCC cell growth and migration. Our findings identified a novel CD63-IL-6/IL-27-STAT3 axis in the development of HCC and provided a potential target for the diagnosis and treatment of this disease.     

PD-L1和CD147在口腔鳞癌中的表达与临床意义

目的:探讨口腔鳞癌组织中免疫共刺激分子PD-L1与细胞外基质蛋白酶诱导因子CD147的表达、两者的相关性及临床意义。方法:应用免疫组化技术检测66例口腔鳞癌组织及36例正常口腔黏膜组织中PD-L1和CD147的表达,分析PD-L1、CD147表达的相关性及二者与口腔鳞癌临床病理参数的关系。结果:PD-L1在口腔鳞癌组织中表达阳性率为68.18%(45/66),正常口腔黏膜组织中表达阳性率仅为16.67%(6/36);CD147在口腔鳞癌组织中表达阳性率为74.24%(49/66),明显高于其在正常口腔黏膜组织中的表达13.88%(5/36)。PD-L1和CD147两者在口腔鳞癌组织中阳性表达率与口腔黏膜组织相比均明显升高(P0.01)。统计学分析显示,PD-L1和CD147在口腔鳞癌组织中的高表达与患者的性别年龄、吸烟史及肿瘤的体积等因素无明显相关,但与TNM分期及鳞癌的组织分化程度紧密相关。口腔鳞癌组织中PD-L1与CD147两者相关性分析r=0.342,P值小于0.01,说明二者的表达呈显著正相关。结论:口腔鳞癌组织中PD-L1与CD147均呈高表达,并且二者的过度表达可能与口腔鳞癌的发生、发展关系密切,合并检测二者可能为OSCC的诊疗及预后指明新的方向,为口腔鳞癌的靶向治疗提供新的靶点。     

Silencing of CD147 inhibits hepatic stellate cells activation related to suppressing aerobic glycolysis via hedgehog signaling

Hepatic stellate cells (HSCs) activation is a key step that promotes hepatic fibrosis. Emerging evidence suggests that aerobic glycolysis is one of its important metabolic characteristics. Our previous study has reported that CD147, a glycosylated transmembrane protein, contributes significantly to the activation of HSCs. However, whether and how it is involved in the aerobic glycolysis of HSCs activation is unknown. The objective of the present study was to validate the effect of CD147 in HSCs activation and the underlying molecular mechanism. Our results showed that the silencing of CD147 decreased the expression of α-smooth muscle-actin (α-SMA) and collagen I at both mRNA and protein levels. Furthermore, CD147 silencing decreased the glucose uptake, lactate production in HSCs, and repressed the lactate dehydrogenase (LDH) activity, the expression of hexokinase 2 (HK2), glucose transporter 1 (Glut1). The effect of galloflavin, a well-defined glycolysis inhibitor, was similar to CD147 siRNA. Mechanistically, CD147 silencing suppressed glycolysis-associated HSCs activation through inhibiting the hedgehog signaling. Moreover, the hedgehog signaling agonist SAG could rescue the above effect of CD147 silencing. In conclusion, CD147 silencing blockade of aerobic glycolysis via suppression of hedgehog signaling inhibited HSCs activation, suggesting CD147 as a novel therapeutic target for hepatic fibrosis.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10616-021-00460-9.     

The expression and significance of NET-1 and Contactin in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and a major cause of cancer-related mortality. In this study, the significance of NET and Contactin on the pathogenesis and prognosis of HCC was investigated, and further to explore their functions in vitro and in vivo by down regulation with siRNA. The expression of NET-1 and Contactin in HCC and in adjacent non-tumor tissues (ANT) were evaluated by immunohistochemistry, and the correlations of the expression of NET-1 and Contactin with the clinicopathological characteristics and survival of HCC patients were also analyzed. After inhibited by single-target siRNA or dual-target siRNA, the expressions of NET-1 and Contactin mRNA and protein in SMMC-7221 cells were determined by RT-PCR, Western blot and immunofluorescence stain. The cell proliferation and apoptosis were assessed by CCK-8 assays and flow cytometry (FCM). The ability of cell migration and invasion were evaluated by wound-healing migrating assay and transwell chamber assays, respectively. Subsequently, transmission system encapsulated cationic liposome was used to deliver dual-siRNA into HCC xenografts in mice. The expressions of NET-1, Cortactin, Ki67, Bax, Bcl2 and Survivin in xenograft tumor were detected by immunohistochemical staining, respectively. The positive rates of NET-1 and Cortactin in HCC tissues were significantly higher than those in ANT. In HCC, the expression of NET-1 was related to Edmondson''s grade (P<0.05), cirrhosis background (P<0.001) and TNM stage (P<0.05). The expression of Cortactin was related to tumor infiltration (P<0.05), vascular invasion (P <0.001) and TNM stage (P<0.001). The expressions of NET-1 and Cortactin were positively correlated (r=0.280, P=0.004). Significant differences in the 5-year survival rates were seen between the NET-1 negative group and the positive group (P<0.05), and between the Contactin negative group (50%) and Contactin positive group (28.0%, P<0.01). The 5-year overall survival rate (OS) in NET-1 and Contactin co-expression cases (27.78%) were remarkably lower than that in both NET-1 and Contactin negative cases (54.54%) and in NET-1 positive while Contactin negative cases (47.06%, P<0.05). Univariate and multivariate Cox regression analysis revealed that NET-1 and Contactin over-expression were independent indicators for OS in HCC patients (P<0.01). There were higher expressions of NET-1 and Contactin in SMMC-7721 cells than that in other HCC cells. Dual-siRNA was demonstrated to be more effective on inhibiting cancer cell proliferation, migration and inducing apoptosis than individual siRNAs used alone in vitro and in vivo (P<0.05). The results suggest that dual-siRNA may be a great potential in siRNA-based therapeutic applications.     

A novel microRNA signature predicts vascular invasion in hepatocellular carcinoma

Vascular invasion (VI) in hepatocellular carcinoma (HCC) is an important clinical parameter to predict survival. In this study, we collected microRNA (miRNA) expression data from HCC patients using The Cancer Genome Atlas database and identified a novel miRNA signature associated with VI. First, we categorized HCC patients into groups with or without VI (VI+ and VI−). We identified three miRNAs (miRNA-210, miRNA-10b, and miRNA-9-1) that were associated with VI according to a Kaplan–Meier analysis. This three-miRNA signature exhibited good predictive ability for VI in patients with HCC according to a receiver operating characteristic curve analysis at 1, 3, and 5 years. Patients with HCC with a high risk score exhibited a trend toward worse outcomes as determined by multivariable Cox regression and stratified analyses. This three-miRNA signature provides an accurate prediction of VI and can be used as an independent prognostic indicator for predicting VI in HCC patients.     

Blocking CD147 induces cell death in cancer cells through impairment of glycolytic energy metabolism

CD147 is a multifunctional transmembrane protein and promotes cancer progression. We found that the anti-human CD147 mouse monoclonal antibody MEM-M6/1 strongly induces necrosis-like cell death in LoVo, HT-29, WiDr, and SW620 colon cancer cells and A2058 melanoma cells, but not in WI-38 and TIG-113 normal fibroblasts. Silencing or overexpression of CD147 in LoVo cells enhanced or decreased the MEM-M6/1 induced cell death, respectively. CD147 is known to form complex with proton-linked monocarboxylate transporters (MCTs), which is critical for lactate transport and intracellular pH (pHi) homeostasis. In LoVo cells, CD147 and MCT-1 co-localized on the cell surface, and MEM-M6/1 inhibited the association of these molecules. MEM-M6/1 inhibited lactate uptake, lactate release, and reduced pHi. Further, the induction of acidification was parallel to the decrease of the glycolytic flux and intracellular ATP levels. These effects were not found in the normal fibroblasts. As cancer cells depend on glycolysis for their energy production, CD147 inhibition might induce cell death specific to cancer cells.