A new method for assaying adhesion of cancer cells to the greater omentum and its application for evaluating anti-adhesion activities of chemically synthesized oligosaccharides

A new ex vivo method for assaying adhesion of cancer cells to the greater omentum has been developed using mouse greater omentum and [³H]labelled human gastric and mouse colorectal cancer cells. Since the adhesion rates were found to increase up to 18 h and labelled cells seemed to be stable during the period, the present method could be useful for investigating adhesion of cancer cells to the greater omentum, which must occur at the first step of the peritoneal dissemination. The adhesion of cancer cells to the greater omentum was inhibited by a series of chemically synthesized oligosaccharides and Galβ1,3[3OMeGalβ1,4GlcNAcβ1,6]αBn was found to be the best inhibitor. The anti-tumor effect of this novel tetrasaccharide in vivo was shown in preliminary experiments using Balb/c mice and colon26 cells.     

MicroRNA‐198‐5p inhibits the migration and invasion of non‐small lung cancer cells by targeting fucosyltransferase 8

MicroRNA‐198‐5p (miR‐198‐5p) displays crucial roles in various cancers including non‐small cell lung cancer (NSCLC), but the underlying molecular mechanisms remain unclear. Fucosyltransferase 8 (FUT8) is associated with tumour metastasis and prognosis. In this study, we explored the expression of miR‐198‐5p and FUT8 in NSCLC patients. Results showed that miR‐198‐5p was under‐expressed in NSCLC tissues and was negatively correlated with tumour size, lymph node metastasis and tumour‐node‐metastasis stage, while FUT8 expression was highly upregulated. Next, we altered miR‐198‐5p expression using the mimic or inhibitor in the functional study. Results showed that miR‐198‐5p overexpression could inhibit the migration, invasion and epithelial‐to‐mesenchymal transition (EMT) of NSCLC cells; reversely, suppression of miR‐198‐5p enhanced cell migration, invasion and EMT. In vivo, miR‐198‐5p overexpression inhibited the formation of mouse lung and liver metastasis. Luciferase reporter, real‐time PCR and western blot assays showed that miR‐198‐5p could directly target FUT8 and regulate FUT8 expression. Further, FUT8 overexpression reversed the effect of miR‐198‐5p overexpression on the migration, invasion and EMT of NSCLC cells. Taken together, miR‐198‐5p functions as a tumour suppressor by targeting FUT8 in NSCLC. MiR‐198‐5p may be developed as a new diagnostic biomarker and therapeutic target for lung cancer.     

中国福建地区类孟买血型个体Fut1基因突变研究

本研究旨在探讨中国福建地区类孟买血型的分子机制。采用血清学方法鉴定研究对象为类孟买血型,采用PCR扩增研究对象的α1,2岩藻糖基转移酶(FUT1)基因编码区序列,并将PCR产物经TA后进行测序,分析fut1基因编码区序列,以证实类孟买血型的发生机制。结果表明,PCR扩增产物电泳分析证实成功扩增fut1基因编码区序列;克隆后测序分析发现本例先证者2条染色体上fut1基因均为第547-552位AG两碱基缺失(CAGAGAG→CAGAG),导致阅读框架发生移码,提前形成终止密码。结论:本例类孟买血型个体fut1基因为h1h1型纯合突变。     

FUT1 基因杂合或纯合突变致类孟买血型形成研究简

本研究旨在探讨类孟买血型表型形成的分子机制.采用血清学方法鉴定个体红细胞H抗原;采用高保真PCR扩增检测个体的α1,2岩藻糖基转移酶（FUT1）基因编码区序列,并进行测序、比对分析,以查明个体类孟买血型的发生机制.结果表明：凝胶电泳分析证实成功扩增FUT1基因编码区全长序列;克隆后测序、比对发现：个体1的1条染色体上FUT1基因为h1 （547-552delAG）,另1条染色体上为h4（35C＞ T）,为h1h4杂合子;个体2,3的2条染色体上FUT1基因均为h1 （547-552delAG）,为h1h1纯合子.结论：FUT1基因杂合或纯合突变均可致类孟买血型的发生.     

Comparative analysis of three genetic modifications designed to inhibit human serum‐mediated cytolysis

Hyperacute rejection (HAR) remains a critical immunologic hurdle in the development of xenogeneic organs for human transplantation. Strategies that simultaneously eliminate both natural antibody reactivity and complement activation on the xenogeneic cell surface may be the best approach to achieve clinical application of xenogeneic vascularized organ transplantation. We have developed multiple lines of genetically manipulated mice to evaluate the combination of different genetic approaches aimed at inhibiting antibody and complement-mediated cell lysis. We utilized transgenic mice expressing the human complement inhibitor, CD59, the human 1,2-fucosyltransferase (H-transferase, HT) and the α1,3-galactosyltransferase (α1,3-GT) knock-out mouse line (Gal KO). Our data show that expression of hCD59 in combination with HT expression or the null phenotype of α1,3-GT are equally effective at preventing human serum-mediated cytolysis. Interestingly, the triple combination affords no additional protective effect. Therefore, coexpression of HT and a complement inhibitor is the most immediate strategy to genetically engineer transgenic pigs to be used as xenogeneic donors.     

Tumor cells as the origin of elevated serum α1,3fucosyltransferase in association with malignancy

We have previously reported that the elevated activities of serum α1,3fucosyltransferase reverted to normal levels after curative removal of the tumors. To determine the origin of elevated serum α1,3fucosyltransferase, blood samples were obtained from both the drainage vein and the artery in patients with different stages of colorectal cancer at surgery. The enzyme levels in all samples from the drainage vein were found to be higher than the levels in the artery that fed the tumor. Hence, the origin of elevated α1,3fucosyltransferase in serum was thought to be the tumor rather than the liver that is the normal source of serum α1,3fucosyltransferase. When serum samples not only from colorectal cancer patients but also from patients with gastric, liver, lung, pancreas, bladder and esophagus cancer were treated with anti-FUTVI antibody, the measured activities of α1,3fucosyltransferase were markedly reduced. Further, secretion of α1,3fucosyltransferase from human colorectal carcinoma cells was also detected in the culture medium by Western immuno-blot analysis with anti-FUTVI antibody. This revised version was published online in July 2006 with corrections to the Cover Date.     

Human fucosyltransferase 6 enables prostate cancer metastasis to bone

Background:

The interaction between human prostate cancer (PCa) cells and bone marrow (BM) endothelium follows a rolling-and-adhesion cascade mediated by E-selectin ligand (ESL): E-selectin. This adhesion is enabled by elevated expression of α-1,3-fucosyltransferases (FTs), enzymes responsible for ESL-mediated bone metastasis in humans. In contrast, the incidence of bone metastasis in mice is rare.

Methods:

FT 3, 6 and 7 were overexpressed in mouse PCa cells. The rolling cell number, cell-rolling velocity and transendothelial migration were characterised in vitro. Fucosyltransferases-transduced mouse PCa cells expressing luciferase were inoculated into mice via left ventricle to compare the capability of bone metastasis. Mass spectrometry and immunoprecipitation were utilised for identification of ESLs.

Results:

Overexpression of FT3, FT6 or FT7 restored ESLs and enabled mouse PCa cells to roll and adhere in E-selectin-functionalised microtubes, similar to trafficking of circulating PCa cells in BM vessels. Following intracardiac inoculation, FT6-transduced cells induced robust bone metastasis in mice. Inhibition of FT6 by a fucose mimetic significantly reduced bone metastasis. Importantly, comparison of FT3, FT6 and FT7 gene expression in existing clinical samples showed significant upregulation of FT6 in PCa-distant metastases.

Conclusion:

FT6 is a key mediator of PCa cells trafficking to the BM. It may serve as a viable drug target in preclinical tests of therapeutics for reduction of PCa bone metastasis.     

Downregulation of microRNA‐15b by hepatitis B virus X enhances hepatocellular carcinoma proliferation via fucosyltransferase 2‐induced Globo H expression

Globo H, a cancer‐associated carbohydrate antigen, is highly expressed in various types of cancers. However, the role of Globo H in hepatocellular carcinoma (HCC) remains elusive. In our study, we performed glycan microarray analysis of 134 human serum samples to explore anti‐Globo H antibody changes and found that Globo H is upregulated in hepatitis B virus (HBV)‐positive HCC. Similarly, immunohistochemistry showed that Globo H expression was higher in tumors compared to normal tissues. In addition, fucosyltransferase 2 (FUT2), the main synthetic enzyme of Globo H, was also increased in HCC cells overexpressing HBV X protein (HBX). HBX plays an important role in promoting cell proliferation and may be related to increased levels of FUT2 and Globo H. Furthermore, using microRNA profiling, we observed that microRNA‐15b (miR‐15b) was downregulated in patients with HCC and confirmed association of FUT2 expression with expression of its product, Globo H. Therefore, our results suggest that HBX suppressed the expression of miR‐15b, which directly targeted FUT2 and then increased levels of Globo H to enhance HCC cell proliferation. Additionally, proliferation of HBX‐overexpressing HCC cells was significantly inhibited by treatment with Globo H antibody in vitro. In xenograft animal experiments, we found that overexpression of miR‐15b effectively suppressed tumor growth. The newly identified HBX/miR‐15b/FUT2/Globo H axis suggests one possible molecular mechanism of HCC cell proliferation and represents a new potential therapeutic target for HCC treatment.     

岩藻糖转移酶9在乳腺癌中的表达及其临床意义

目的 本研究探讨岩藻糖转移酶9（fucosyltransferase9,FUT9）在乳腺癌组织中的表达及其与乳腺癌临床病理因素的关系。方法 采用免疫组织化学SP法检测11例良性乳腺增生组织和114例乳腺癌组织（其中47例合并转移淋巴结,67例无转移淋巴结）中FUT9的表达。结果 FUT9在良性乳腺增生组织中的阳性表达率为9.1%（1/11）,而在乳腺癌组织中的阳性表达率为71.9%(82/114),在转移的淋巴灶中阳性表达率为100%（47/47）。乳腺癌组织中的阳性表达率明显高于良性乳腺增生组织的阳性表达率,两组比较差异有统计学意义（P<0.01）。乳腺癌组织中有淋巴结转移组的阳性表达率为80.9%（38/47）,明显高于无淋巴结转移组的阳性表达率65.7%（44/67）,两组比较差异有统计学意义（P<0.05）。在有淋巴结转移组FUT9的表达与TNM分期以及转移的淋巴结个数相关（P<0.05）。FUT9的表达与ER、PR、HER2的表达无相关性（P>0.05）。结论 FUT9在乳腺癌组织中的表达明显增高;FUT9的表达与淋巴结转移、转移的淋巴结个数以及TNM分期有关;FUT9可能与乳腺癌淋巴道转移有关。     

岩藻糖基转移酶II对前列腺癌细胞生物学功能的影响及机制

目的：探讨岩藻糖基转移酶II（FUT2）对前列腺癌细胞生物学功能的影响及其潜在机制。方法：通过免疫组化技术检测FUT2 在前列腺癌组织中的表达情况。利用脂质体转染技术构建低表达FUT2的前列腺癌细胞株及其对照细胞株,并通过MTT实验、平板克隆实验、Transwell实验检测干扰FUT2表达对前列腺癌细胞生物学功能的影响。采用Western blot检测β-catenin、Cyclin D1和ICAM-1等蛋白表达水平。结果：FUT2在前列腺癌组织中的表达水平明显升高（P ＜0.05）。干扰FUT2的表达后,前列腺癌细胞的增殖、迁移和侵袭能力下降,Cyclin D1表达下调,ICAM-1表达上调（P ＜0.05）。结论：FUT2促进前列腺癌细胞的增殖、迁移和侵袭能力。