High level of Nm23-H1 gene expression is associated with local colorectal cancer progression not with metastases.

This study aimed to determine the expression of Nm23-H1 in colorectal cancer and liver metastases and to correlate Nm23-H1 expression with clinicopathological variables. Specimens from 59 primary colorectal cancers and five liver metastases were studied using Northern blot hybridisation. The mean +/- s.e. of tumour/normal (T/N) ratio of Nm23-H1 RNA expression was 4.3 +/- 0.4 (P < 0.001) and 5.1 +/- 0.90 (P < 0.01) for colorectal cancer and liver metastases respectively. No significant relationship was observed between the level of Nm23-H1 RNA and the patient''s age, sex, tumour location, differentiation, presence of lymph node involvement or distant metastases. Nm23-H1 RNA level was 2.6 +/- 0.5 for tumour size less than 3.0 cm and 4.6 +/- 0.5 for those > or = 3.0 cm (P = 0.05). There appeared to be a trend between increasing relative Nm23-H1 RNA and bowel wall invasion, irrespective of metastatic status (T1 = 1.9 +/- 0.3, T2 = 4.1 +/- 0.6, T3 = 4.1 +/- 0.5 and T4 = 6.4 +/- 1.6). This difference was statistically significant when T1 was compared against > or = T2 lesions (P = 0.01). Western blot analysis reveals two Nm23H-1 bands (17.0 kDa and 18.5 kDa). In 16 colorectal patients, the T/N fold-increase in protein expression was 2.66 +/- 0.46 (P < 0.001) and 2.40 +/- 0.32 (P < 0.001) for the 17.0 and 18.5 kDa band respectively. Both Nm23-H1 RNA and protein levels in primary colorectal cancers do not appear to correlate with synchronous regional or distant metastases. Since Nm23-H1 RNA expression is associated with increasing tumour size and tumour local invasion, Nm23-H1 RNA expression may be associated with local disease progression.     

Nm23-H1 expression does not predict clinical survival in colorectal cancer patients

The gene Nm23, which encodes for a nucleoside diphosphate kinase, has been defined as a metastasis-suppressor gene because of the inverse correlation between its expression and the metastatic capacity of the tumor cells. For colorectal cancer, however, the findings are equivocal. The aim of our study was to assess, in 160 patients undergoing surgery for colorectal cancer (CRC), the expression of the Nm23-H1 protein and to evaluate its possible associations with traditional clinicopathologic variables, with DNA-ploidy and proliferative activity (S-phase fraction, SPF), and with disease-free and overall survival of patients. Nm23-H1 expressions were evaluated on paraffin-embedded tissue by immunohistochemistry; DNA-ploidy and SPF on frozen tissue by flow-cytometric analysis. The median follow-up time in our study group was 71 months (range 34-115 months). No association was observed between Nm23-H1 protein expression and clinicopathological variables, S-phase fraction and DNA-ploidy. Furthermore, no significant differences were observed in the survival of patients with either moderate or strong Nm23-H1 expression. The major significant predictors for both disease relapse and death were advanced Dukes' stage, DNA aneuploid tumors and high SPF, while lymphohematic invasion was the only independent factor for relapse and non-curative resection for death. Our results indicate that Nm23-H1 activity is tissue-specific and that in CRCs the expression of the protein is not associated with tumor progression and patient prognosis, although further studies are required in order to throw more light on the possible clinical significance of the overexpression of the protein Nm23-H1 in such tumors.     

nm23-H1的表达和突变与卵巢癌浸润转移的关系

 目的：探讨卵巢肿瘤组织中nm23－H1的表达和突变与其浸润转移的关系。方法：采用免疫组织化学,反转录 多聚酶链反应（RT－PCR）和多聚酶链反应 单链构象多态技术（PCR SSCP）方法检测41例卵巢癌,20例卵巢良性肿瘤和21例正常卵巢组织nm23－H1的表达和突变。结果：（1）卵巢癌组织中V阳性表达率明显高于良性卵巢肿瘤及正常对照（P＜0.01）;（2）卵巢癌灶nm23－H1的阳性表达及扩增明显高于转移灶（P＜0.0）;（3）Ⅰ～Ⅱ期患者癌组织中nm23－H1的阳性表达率明显高于Ⅲ～Ⅳ期者（P＜0.0）;（4）nm23－H1的阳性表达和扩增与病理分级及病理分类无显著差别（P＞0.05）;（5）nm23－H1阳性表达的患者近期疗效比阴性者好（P＜0.05）。（6）卵巢癌组织中nm23－H1的突变率为7.3%且均发生在晚期患者,而良性肿瘤和正常对照未发现有突变。结论：nm23－H1阳性表达与卵巢癌的浸润转移有关,对它的检测有助于临床上判断预后。     

Inverse association of nm23-H1 expression by colorectal cancer with liver metastasis.

The expression of nm23-H1 mRNA and protein was studied in colorectal cancers by Northern blotting and immunohistochemistry. All 21 colorectal cancers studied by Northern blotting had increased levels of nm23-H1 mRNA relative to the adjacent normal colonic mucosa. Increased nm23-H1 protein expression was also observed in all 36 colorectal cancer cases including those studied by Northern blotting. There was no significant correlation between nm23-H1 expression and tumour histology, serosal invasion, lymphatic invasion, venous invasion, or lymph node metastasis. However, the expression of both mRNA and protein was significantly lower in tumours associated with liver metastasis than in those without such metastasis. These observations indicate that the nm23 gene may play a role in the suppression of liver metastasis of colorectal cancer.     

nm23-H1与大肠癌微转移关系的研究

目的探讨nm23-H1基因表达与大肠癌浸润、微转移和预后的关系。方法经病理确诊的120例大肠癌组织标本,采用SP法检测nm23-H1基因蛋白表达、巢式RT-PCR方法检测进入外周静脉血的大肠癌微转移灶(CK20mRNA),并与同一患者大肠癌组织标本中nm23-H1基因表达情况进行综合分析。结果nm23-H1在大肠癌中的阳性表达率为61.7%(74/120),低分化腺癌中nm23-H1阳性表达率(41.0%)显著低于高中分化腺癌中的表达率(70.3%)(P<0.05);有淋巴结转移者nm23-H1阳性表达率为53.7%,无淋巴结转移者为71.7%,两组比较有显著性差异(P<0.05);120例大肠癌患者术后3年随访中,CK20mRNA阳性检出率为41.5%,nm23-H1阳性表达率与CK2OmRNA阳性率呈显著负相关(P<0.001);nm23-H1蛋白阳性表达组术后3年生存率为60.8%(45/74),阴性表达组术后3年生存率为21.7%(10/46),二组比较有显著性差异(P<0.001)。结论nm23-H1与大肠癌细胞分化、淋巴结转移及外周静脉血中微转移密切相关,是影响大肠癌预后的重要指标之一。     

Clinical-translational approaches to the Nm23-H1 metastasis suppressor

Nm23-H1 significantly reduces metastasis without effects on primary tumor size and was the first discovered metastasis suppressor gene. At least three mechanisms are thought to contribute to the metastasis-suppressive effect of Nm23-H1: (a) its histidine kinase activity toward ATP-citrate lyase, aldolase C, and the kinase suppressor of ras, with the last inactivating mitogen-activated protein kinase signaling; (b) binding proteins that titer out "free" Nm23-H1 and inhibit its ability to suppress metastasis; and (c) altered gene expression downstream of Nm23-H1, particularly an inverse association with the lysophosphatidic acid receptor endothelial differentiation gene-28 (EDG2). Most metastasis suppressor genes, including Nm23-H1, affect metastatic colonization, which is the outgrowth of tumor cells in distant locations; therefore, they are of high translational interest. A phase II trial is ongoing to test the hypothesis that a compound, high-dose medroxyprogesterone acetate (MPA), used as an unconventional gluocorticoid, will stimulate breast cancer cells to reexpress Nm23-H1 and limit subsequent metastatic colonization.     

The Nm23-H1 metastasis suppressor as a translational target

Nm23 was the first of what has become a field of over 20 known metastasis suppressor genes (MSGs). Since the discovery of Nm23 in 1988, a variety of mechanisms have been attributed to its activity, including a histidine kinase activity, binding of other proteins to regulate metastatic formation, and altered gene expression downstream of Nm23. Here, we will review current efforts to translate the previous work done on this MSG into the clinic, including high-dose medroxyprogesterone acetate (MPA), which has been shown to upregulate Nm23 expression. In addition, we will detail a new potential target downstream of Nm23. LPA1 is one of a group of known cell surface receptors for lysophosphatidic acid (LPA), which has been shown to be inversely correlated with Nm23 expression. A specific LPA1 antagonist could conceivably mimic the effects of Nm23 by downregulating the activity of the LPA1 pathway, which would be of considerable interest for potential clinical use.     

Expression of nm23-H1 and nm23-H2 protein in endometrial carcinoma.

nm23 gene expression has been shown to be inversely correlated with tumour metastatic potential in some cancers but not in others. Examination was made of the expression of nm23-H1 and nm23-H2 gene products by immunohistochemistry and immunoblotting in 28 endometrial carcinomas. Immunohistochemistry indicated the cytoplasm of cancer cells to be positive, and myometrium and endometrial stromal cells negative, for nm23-H1 and -H2 protein. The staining intensity for these proteins was significantly stronger in well-differentiated adenocarcinomas (G1) than in those moderately differentiated (G2) (P < 0.05). nm23-H1 and -H2 proteins were shown by immunoblotting to be present at significantly higher levels in G1 than in G2 tumours (P < 0.05). Two of eight cases expressed high nm23-H1 and -H2 protein in poorly differentiated adenocarcinomas (G3). In G3 tumours, nm23 expression may be diverse. In this study, the expression of nm23-H1 and -H2 was not correlated with stage, metastasis, tumour size, myometrial invasion, oestrogen receptor, progesterone receptor or menopause. It follows from the findings presented above that the high expression of nm23-H1 and -H2 is positively correlated with histological differentiation.     

nm23-H1基因表达与卵巢癌转移的相关性

背景与目的：转移是卵巢癌治疗失败及患者死亡的首要原因。然而,目前对卵巢癌转移潜能的分子机制知之不多。本研究旨在筛选高频转移卵巢恶性肿瘤细胞,分析卵巢癌高频转移细胞模型中nm23．H1基因表达与肿瘤转移特性的相关性,为系统实验研究和临床实践提供依据。方法：通过反复动物接种和体外培养,观察动物肺转移状况,筛选高频转移细胞株,比较原发肿瘤和转移肿瘤的特征,并应用Northem blot和Westem blot方法测定各类肿瘤细胞nm23 mRNA和蛋白表达水平。结果：8株卵巢恶性肿瘤细胞中,4株有较高转移潜能,多次培养接种可筛选出高频转移细胞亚群。各类细胞nm23mRNA和蛋白表达水平与肿瘤转移特性呈负相关(r=0．96,P=0．0001)。结论：由基因分子水平决定的肿瘤转移趋势在不同肿瘤种类及细胞亚群中有明显差异;卵巢癌中nm23 mRNA和蛋白的表达与其转移能力的降低有密切关系,可作为判定卵巢癌预后的敏感指标。     

Nm23-H1 modulates the activity of the guanine exchange factor Dbl-1

Cytoskeleton rearrangement is necessary for tumor invasion and metastasis. Cellular molecules whose role is to regulate components of the cytoskeletal structure can dictate changes in cellular morphology. One of these molecules is the suppressor of tumor metastasis Nm23-H1. The level of Nm23-H1 expression has been linked to the invasiveness and metastatic potential of human cancers including melanoma and breast cancer. In this report, we demonstrate an interaction between the suppressor of tumor metastasis Nm23-H1, and Dbl-1, an oncoprotein which is associated with guanine exchange and belongs to a family of Guanine Exchange Factors (GEF). Nm23-H1 also was shown to bind pDbl which is the proto-oncoprotein of Dbl. Interestingly, the interaction between Nm23-H1 and Dbl-1 rescues the suppression of the cell motility activity Nm23-H1. Moreover, this interaction results in loss of the ability of the Dbl-1 oncoprotein to function as a GEF for the critical Rho-GTPase family member Cdc42. The loss of GTP loading onto Cdc42 resulted in a dramatic reduction in adhesion stimulated ruffles and suggests that Nm23-H1 can negatively regulate cell migration and tumor metastasis by modulating the activity of Cdc42 through direct interaction with Dbl-1.     

Expression of thrombospondin-1 in resected colorectal liver metastases predicts poor prognosis.

PURPOSE: The aim of this study was to examine the expression and prognostic relevance of thrombospondin-1 (TSP-1) in tumor biopsies taken from a consecutive series of liver resections done at the University Hospitals of Leicester and the Royal Liverpool Hospital. EXPERIMENTAL DESIGN: Patients having undergone a liver resection for colorectal liver metastases at our institutions between 1993 and 1999 inclusive were eligible. Inclusion criteria were curative intent, sufficient tumor biopsy, and patient follow-up data. One hundred eighty-two patients were considered in this study. Standard immunohistochemical techniques were used to study the expression of TSP-1 in 5-microm tumor sections from paraffin-embedded tissue blocks. TSP-1 was correlated with survival using the Kaplan-Meier method and log-rank test for univariate analysis and the Cox proportional hazard model for multivariate analysis. RESULTS: One hundred eighty-two patients (male, n = 122 and female, n = 60) ages between 25 and 81 years (mean, 61 years) were included. TSP-1 was expressed around blood vessels (n = 45, 25%) or in the stroma (n = 59, 33%). No expression was detected in the remaining tumors. TSP-1 significantly correlated with poor survival on univariate (P = 0.01 for perivascular expression and P = 0.03 for stromal expression) and multivariate analysis (P = 0.01 for perivascular expression). CONCLUSION: TSP-1 is a negatively prognostic factor for survival in resected colorectal liver metastases.     

PTEN、nm23-H1和Syk在大肠癌中的表达及意义

目的 检测PTEN、nm23-H1和Syk蛋白在大肠癌中的表达,探讨其与大肠癌发生、发展、浸润和转移的关系。方法 采用免疫组化SP法检测60例大肠癌组织及30例正常大肠黏膜中PTEN、nm23-H1和Syk的表达,分析三者与大肠癌临床病理特征的关系。结果 正常大肠黏膜中PTEN、nm23-H1和Syk的阳性表达率分别为90.0％（27／30）、93.3％（28／30）和96.7％（29／30）,而在大肠癌组织中分别为36.7％（22／60）、41.7％（25／60）和31.7％（19／60）,差异均有统计学意义（P＜0.01）。大肠癌中3种蛋白的表达与年龄、性别、肿瘤大小和发生部位均无关（P＞0.05）,PTEN、Syk表达与大肠癌的浸润程度、淋巴结转移、分化程度及Duke's分期有关,nm23-H1表达与浸润程度、淋巴结转移及Duke's分期有关。大肠癌中3种蛋白表达两两呈正相关。结论 PTEN、nm23-H1和Syk表达与大肠癌发生、发展、浸润及转移密切相关,联合检测是判断大肠癌生物学行为的重要指标。     

Nm23-H1、CD44v6在60例下咽癌组织中的表达及临床意义

摘 要：［目的］分析下咽癌组织中肿瘤抑制基因 nm23-H1和转移相关基因 CD44v6的表达情况及其意义。［方法］ 60例下咽癌（其中30例有颈部淋巴结转移,30例无颈部淋巴结转移）及30例癌旁组织纳入分析,采用免疫组织化学SP法检测nm23-H1和CD44v6的表达情况。［结果］ Nm23-H1在下咽癌组的表达阳性率为53.33%,较癌旁组织组（90.00%）明显偏低(P＜0.05);有颈部淋巴结转移下咽癌组（40.00%)、无颈部淋巴结转移下咽癌组(66.67%)及癌旁组织各组间nm23-H1阳性率差异有统计学意义 (P＜0.05)。CD44v6在下咽癌组的表达阳性率为76.67%,较癌旁组（56.67%）明显高(P＜0.05);有颈部淋巴结转移组CD44v6阳性率(90.0%)较无转移组(63.33%)明显高(P＜0.05)。下咽癌组织中nm23-H1和CD44v6表达呈负相关(P＜0.05)。［结论］ 下咽癌中 nm23-H1表达下调可促使下咽癌发生、推动肿瘤发展及发生颈部淋巴结转移;CD44v6表达上调可使肿瘤的发展及转移加速。两者的表达失衡与下咽癌发生颈部淋巴结转移关系密切并且可能起协同作用。因此,联合检测nm23-H1和CD44v6对判断下咽癌的颈部淋巴结转移有指导意义。     

Nm23 and breast cancer metastasis

Summary The majority of breast cancer patients succumb to metastatic disease. We summarize published and recent research concerning thenm23 gene in breast cancer metastasis. In a murine developmental study, nm23 expression increased with the functional differentiation of the mammary gland in nulliparous and pregnant animals. In human breast cancer, five studies have now demonstrated a significant association between reduced nm23 expression, at the RNA or protein levels, and aggressive tumor behavior. Nm23-negative tumor cells have been observed in comedo ductal carcinoma in situ lesions in two independent studies, indicating that decreases innm23 expression begin prior to actual histologically identifiable invasion. Transfection studies, in which humannm23-H1 cDNA was expressed in the metastatic human MDA-MB-435 breast carcinoma cell line, indicate thatnm23-H1 suppresses in vivo metastatic potential by 50-90%. Finally, our data in melanoma and breast carcinoma transfection systems suggest that the biochemical mechanism of nm23 suppressive activity is likely not due to its nucleoside diphosphate kinase activity, association with GAP proteins, or secretion from cells.Supported by an educational grant from CIBA-GEIGY Corp., at the San Antonio Breast Cancer Symposium, Dec. 8, 1992.     

nm23—H1基因在大肠癌中的表达与肝转移及预后的关系

目的：探讨ｎｍ２３－Ｈ１蛋白表达与大肠癌肝转移及预后的关系。方法：对１０１例大肠癌存档石蜡块进行重新切片，采用ｎｍ２３－Ｈ１单克隆抗体进行免疫组化染色（ＬＳＡＢ法）。结果：ｎｍ２３－Ｈ１蛋白表达与年龄、性别、肿瘤大小、部位、组织类型、浆膜侵犯无关；与Ｄｕｋｅｓ分期、淋巴结转移有关；手术时有肝转移组较无肝转移组低，手术后有肝转移复发组较无肝转移复发组低（Ｐ＜００１）。Ｃｏｘ模型分析显示ｎｍ２３－Ｈ１是大肠癌预后的一个保护性指标。结论：ｎｍ２３－Ｈ１基因对大肠癌肝转移和预后具有重要作用。ＬＳＡＢ法检测大肠癌组织中ｎｍ２３－Ｈ１蛋白表达可能是预测大肠癌肝转移及预后的生物学指标之     

联合检测nm23-H1、E-cadherin基因蛋白在鼻咽癌中的表达

目的　探讨 nm2 3- H1基因和粘附分子 E- cadherin在鼻咽癌中的表达情况及其与转移的关系。方法　用免疫组化 SP法检测 nm2 3- H1和 E- cadherin基因产物在 42例鼻咽癌中的表达。结果　 42例鼻咽癌活检标本中 ,nm2 3- H1蛋白表达阳性率 5 4 .7% ,E- cadherin表达阳性率 5 7.1 % ,nm2 3- H1蛋白在有颈淋巴结转移患者中表达阳性率为 40 .7% ,在无颈淋巴结转移患者中表达阳性率为 80 .0 % ,两者差异有显著意义 ( P<0 .0 5 )。E- cadherin在颈淋巴结转移阳性患者中表达阳性率为 44.4% ,颈淋巴结转移阴性患者中表达阳性率为 80 .8% ,两者差异有显著意义 ,( P<0 .0 5 )。两种基因产物表达在鼻咽癌中呈正相关 ( P<0 .0 1 )。结论　nm2 3- H1和 E- cadherin基因与鼻咽癌的转移密切相关 ,对临床判断鼻咽癌预后有一定指导意义