nm23-H1 protein immunohistochemical expression in human breast cancer

A series of 76 patients undergoing surgery for primary breast carcinoma has been prospectively studied in order to evaluate the relative weight of nm23-H1 protein expression in disease-free survival. Expression of nm23 protein was immunohistochemically assessed. In all, 39% (29/74) of the turners showed positive staining for nm23-H1 protein expression. Negative nm23-H1 expression was found in poorly differentiated, tumors (p<0.02). There was no significant relationship between nm23-H1 and the other clinicopathological and biological features examined. In the univariate statistical analysis, node positivity, G3 histological grade and high flow cytometric S phase fraction (SPF) value proved to be significantly related to risk of relapse. In the multivariate analysis, only histological grade (G3) and high SPF values (>10.6) proved to be independently related to risk of relapse, with a hazard ratio of 9.84 and 7.98 respectively. Our preliminary study suggests that immunohistochemical nm23-H1 expression should not be considered a marker for predicting tumor progression and patient prognosis.     

nm23-H1 expression in salivary adenoid cystic carcinoma in relation to metastasis and survival

The expression of nm23-H1, product of putative metastasis suppressor gene, was evaluated immunohistochemically in 31 cases of adenoid cystic carcinoma (ACC) of salivary glands and correlated with their clinicopathologic features. All benign salivary gland tumors of various types, which were used as a non-metastatic control, showed obvious nm23-H1 expression. The immunoreactivity of tumor cells was stronger than that of normal salivary gland components, although the distribution patterns of positive cells considerably varied between tumor types. In ACC, 16 cases (52%) showed the reduction of nm23-H1 immunoreactivity either in positive cell frequency or staining intensity. These cases were referred to as negative cases. The incidence of negative cases was 67% (10/15) and 38% (6/16) of the cases with and without metastasis, respectively. Furthermore, metastatic tumors showed decreased immunoreactivity of this protein compared with their primary tumors. The prognosis of patients with a nm23 negative tumor was generally poorer than that with a positive tumor. These results may suggest that the reduction of nm23-H1 protein has an implication for metastasis of ACC.     

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。     

nm23-H1基因在肺多形性癌组织中的表达及意义

目的　探讨 nm2 3- H1基因表达与肺多形性癌转移及预后的关系。方法　应用免疫组化 S- P法回顾性分析 2 0例肺多形性癌组织中 nm2 3- H1基因表达 ,并结合肺多形性癌的转移、侵袭力、肿块大小及预后进行分析。结果　 nm2 3- H1基因在肺多形性癌中表达阳性率 ,有淋巴结转移组 (2 5 .0 % )低于无淋巴结转移组 (87.5 % ) (P<0 .0 1) ,有侵犯周围组织组 (12 .5 % )低于无侵犯周围组织组 (10 0 % ) (P<0 .0 1) ,肿块直径 >6 cm组 (4 2 .8% )与肿块直径 <6 cm组 (5 0 .0 % ) ,差异无显著性 (P>0 .0 5 )。结论　 nm 2 3- H1基因表达与肺多形性癌有无淋巴结转移、侵袭力强弱关系密切 ,而与肿块大小无关。 nm 2 3- H 1基因高表达预示肺多形性癌转移及侵袭力低 ,预后可能较好     

胃癌nm23基因mRNA原位杂交和免疫组化研究

目的 :探讨胃癌mn2 3 H1基因表达与淋巴结转移倾向的相关性。方法 :采用地高辛标记cD NAnm2 3 H1探针 ,原位杂交检测 2 1例胃癌及癌旁组织 ;应用单克隆抗体 (NM30 1)SP法检测 131例胃癌及 60例非胃癌胃粘膜nm2 3/NDPK表达。结果 :原位杂交阳性率 57.1% ( 12 /2 1) ;免疫组化阳性率为87.8% ( 115/131) ,其中非转移组阳性率为 97.1% ( 34 /35) ,转移组阳性率 84 .4 % ( 81/96) ,P <0 .0 5。结论 :胃癌nm2 3 H1基因表达与淋巴结转移倾向呈负相关 ,与PCNA分级呈正相关     

Identification of a second human nm23 gene, nm23-H2

Reduced RNA and/or protein levels corresponding to the murine nm23-1 and human nm23-H1 complementary DNA clones have been correlated with high tumor metastatic potential in several rodent model systems and human breast carcinomas. We report the identification of a second human nm23 gene, designated nm23-H2. The pNM23-H2S complementary DNA clone predicted a Mr 17,000 protein 88% identical to nm23-H1. nm23-H2 also shared a significant homology with nucleoside diphosphate kinases and a Drosophila developmental gene. Southern blots containing BglII-restricted genomic DNA, which exhibited an allelic restriction fragment length polymorphism for nm23-H1, contained nonallelic bands upon rehybridization to the nm23-H2 probe. Thus, nm23-H1 and nm23-H2 are distinct genes. Northern blot hybridization of nm23-H1- and nm23-H2-specific probes to breast tumors and cell lines indicated that nm23-H1 expression was reduced in high metastatic potential tumor cells to a greater extent than nm23-H2. The data indicate the existence of a family of independently regulated nm23 genes.     

Clinical significance of intracytoplasmic nm23-H1 expression in diffuse large B-cell lymphoma.

PURPOSE: Recently, we established an ELISA technique for measuring nm23-H1 protein in serum and found that the serum nm23-H1 level is a potential prognostic factor for patients with non-Hodgkin's lymphoma. EXPERIMENTAL DESIGN: We used immunohistochemistry to examine the expression of nm23-H1 by the lymphoma cells in patients with diffuse large B-cell lymphoma (DLBCL). RESULTS: By analyzing a consecutive series of 172 untreated DLBCL patients, we found that 100 (58.1%) were strongly positive. The cytoplasmic nm23 expression in lymphoma cells correlated significantly with the serum nm23-H1 level. There was a significant correlation between patients with cytoplasmic nm23-positive lymphoma and those with performance status 2-4, stage III/IV, bulky mass, B symptoms, elevated serum level of soluble interleukin 2 receptor, and elevated serum level of C-reactive protein. Overall and progression-free survival rates were significantly lower in patients with nm23-H1-positive lymphomas than in those with nm23-H1-negative lymphomas. Similar difference was seen between patients with high and low serum levels of nm23-H1. Thus, the correlation between presence or absence of cytoplasmic nm23-H1 expression and serum nm23-H1 levels suggests that serum nm23-H1 is produced directly by lymphoma cells. CONCLUSION: We suggest that nm23-H1 expression is a prognostic factor for DLBCL, and that it is as important as serum nm23-H1, both of which are useful for planning a treatment strategy.     

Evidence for interaction between human PRUNE and nm23-H1 NDPKinase

We have isolated a human and murine homologue of the Drosophila prune gene through dbEST searches. The gene is ubiquitously expressed in human adult tissues, while in mouse developing embryos a high level of expression is confined to the nervous system particularly in the dorsal root ganglia, cranial nerves, and neural retina. The gene is composed of eight exons and is located in the 1q21.3 chromosomal region. A pseudogene has been sequenced and mapped to chromosomal region 13q12. PRUNE protein retains the four characteristic domains of DHH phosphoesterases. The synergism between prune and awdK-pn in Drosophila has led various authors to propose an interaction between these genes. However, such an interaction has never been supported by biochemical data. By using interaction-mating and in vitro co-immunoprecipitation experiments, we show for the first time the ability of human PRUNE to interact with the human homologue of awd protein (nm23-H1). In contrast, PRUNE is impaired in its interaction with nm-23-H1-S120G mutant, a gain-of-function mutation associated with advanced neuroblastoma stages. Consistently, PRUNE and nm23-H1 proteins partially colocalize in the cytoplasm. The data presented are consistent with the view that PRUNE acts as a negative regulator of the nm23-H1 protein. We discuss how PRUNE regulates nm23-H1 protein and postulate possible implications of PRUNE in neuroblastoma progression.     

Reduced expression of nm23-H1, but not of nm23-H2, is concordant with the frequency of lymph-node metastasis of human breast cancer

The nm23 gene is a potential metastasis-suppressor gene originally identified in a murine melanoma line. Several investigators have reported the probable inverse association of nm23 expression with disease prognosis and/or metastasis. Since there are now 2 known isotypes of human nm23, namely nm23-H1 and -H2, we immunohistochemically examined expression of these isotypes in human breast-cancer tissues using monoclonal antibodies (MAbs) specific for each isotype protein. We also analyzed expression of c-erbB-2 in the same collection of cancer tissues, in order to examine the significance of nm23 expression in comparison with c-erbB-2 expression. Of 130 tumors from breast-cancer patients, 73 (56%) and 69 (53%) positively expressed nm23-H1 and -H2 respectively. Expression of c-erbB-2 was positive in 36 (28%). Expression of nm23-H1, but not nm23-H2, was inversely associated with lymph-node metastasis (p < 0.01). Expression of c-erbB-2 was associated with Tnm stage, tumor size and lymph-node metastasis (p < 0.01, p < 0.05 and p < 0.05 respectively). Overall survival was better (p = 0.014) in patients in whom expression of nm23-H1 was positive than in those in whom it was negative. In multivariate analyses using a Cox's proportional-hazards regression model with 9 variables, nm23-H1 showed the fourth greatest contribution to patient survival following lymph-node metastasis, Tnm stage and menopausal status. No significant contribution was shown for c-erbB-2 expression. nm23-H1, but not nm23-H2, may perform a role in disease prognosis in addition to its participation in cancer metastasis. It may have value for predicting long-term survival of human breast-cancer patients. © 1993 Wiley-Liss, Inc.     

Prognostic significance of nm23-H1 expression in oral squamous cell carcinoma

Recent studies indicated nm23-H1 played a role in cancer progression. Therefore, we investigated clinical significance of nm23-H1 expression in oral squamous cell carcinoma (OSCC). In total, 86 OSCC specimens were immunohistochemically stained with nm23-H1-specific monoclonal antibodies. Immunohistochemical staining of nm23-H1 was confirmed by immunoblotting. The relations between nm23-H1 expression and clinicopathologic variables were evaluated by chi(2) analysis. As increased size of primary tumour could escalate metastatic potential and the data of patients at the late T stage might confound statistical analyses, we thus paid special attention to 54 patients at the early T stage of OSCC. Statistical difference of survival was compared by a log-rank test. Immunohistochemically, nm23-H1 expression was detected in 48.8% (42 out of 86) of tumorous specimens. It positively correlated with larger primary tumour size (P=0.03) and inversely with cigarette-smoking habit (P=0.042). In patients at the early T stage, decreased nm23 expression was associated with increased incidence of lymph node metastasis (P=0.004) and indicated poor survival (P=0.014). Tumour nm23-H1 expression is a prognostic factor for predicting better survival in OSCC patients at the early T stage, which may reflect antimetastatic potential of nm23. Therefore, modulation of nm23-H1 expression in cancer cells can provide a novel possibility of improving therapeutic strategy at this stage. In addition, our results further indicated cigarette smoking could aggravate the extent of nm23-H1 expression and possibly disease progression of OSCC patients.     

鼻咽癌组织中nm23-H1和survivin蛋白表达及其临床意义

目的探讨nm23H1和survivin基因蛋白表达与鼻咽癌（NPC）预后的相关性，方法对115例NPC放疗前活检标本，应用免疫组织化学技术检测nm23-H1和surivin基因蛋白在NPC组织中的表达情况，并分析两基因的表达与NPC分期、放疗敏感性、生存率、转移及复发之间的关系。结果NPC组织中nm23-H1和survivin蛋白阳性表达率分别为47．8％和68．7％。NPC分期、淋巴结转移、生存率及远处转移与nm23-H1蛋白低表达有密切关系，而survivin的高表达则与鼻咽癌的颈淋巴结转移、放疗敏感性、生仔率及复发有密切关系。nm23-H1蛋白表达与survivin表达呈负相关（P〈0．05）。结论nm23-H1蛋白低表达和survivin高表达对判断病变进展、预测放疗敏感性、生存率及转移和复发有重要参考价值。     

nm23-H1 expression in squamous cell carcinoma of the head and neck

Archival material from 47 patients with primary squamous cell carcinoma of the head and neck (SCCHN) was studied immunohistochemically for the presence of nm23-H1 protein. Our data indicate that nm23-H1 protein expression is a common event in SCCHN and that there is a trend toward correlation of increased expression of nm23-H1 with increasing tumor size (p = 0.072). The results also show that when adjusting for age and cause of death, there tended to be an inverse relationship between overall survival and the expression of nm23-H1 gene in the primary tumor (p = 0.088).     

肿瘤转移相关基因在甲状腺乳头状癌的表达与颈淋巴结转移相关性研究

目的：探讨肿瘤转移抑制相关基因nm23-H1,转移相关基因CD44v6在甲状腺乳头状癌的表达与颈淋巴结转移的关系。方法：采用免疫组化S－P法,检测201例甲状腺乳头状癌、30例甲状腺良性病变及30例正常甲状腺组织中nm23-H1和CD44v6的表达情况。结果：1）nm23-H1在甲状腺乳头状癌组的表达阳性率62．7％,明显低于良性病变组和正常组（P＜）．01或P＜0．05）。有转移组nm23-H1的阳性率为54．5％,明显低于无转移组（75．0％）,其蛋白表达与颈淋巴结转移呈负相关。2）CD44 v6在甲状腺乳头状癌组的表达阳性率为71．1％,明显高于两对照组（P均＜0．01）。有转移组CD44v6的阳性率为79．3％,明显高于无转移组（58．5％）,其蛋白表达与颈淋巴结转移呈正相关。3）甲状腺乳头状癌中nm23-H1和CD44v6表达呈显著负相关关系（P＜0．01）。nm23-H1阴性表达伴CD44v6阳性表达的患者发生颈淋巴结转移的可能性大。结论：nm23-H1和CD44v6的表达与甲状腺乳头状癌的颈淋巴结转移密切相关,nm23-H1和CD44v6的表达在淋巴结转移中起协同作用,两者的表达失衡可能与甲状腺乳头状癌发生颈淋巴结转移有密切关系。因此,检测nm23-H1和CD44v6可能作为预测甲状腺乳头状癌颈淋巴结转移潜能的有价值的参考指标。     

nm23-H1 expression in nasopharyngeal carcinoma: correlation with clinical outcome

The expression levels of nm23-H1 mRNA and its protein in human nasopharyngeal carcinoma (NPC) were detected to clarify the relationship between nm23-H1 and metastasis and prognosis of patients with NPC. nm23-H1 mRNA expression in fresh tissues from 78 patients with NPC was investigated by in situ hybridization and RT-PCR. Routine labeling streptavidin-biotin immuno-histochemistry with the nm23-H1 murine monoclonal antibody was employed to study the expression of nm23-H1 protein in paraffin-embedded specimens from 231 patients with NPC treated in our hospital. The clinical pathologic data and results of follow-up were collected. Comparisons between expression of nm23-H1 protein or mRNA and clinical outcome were performed using the χ² test. Multivariate prognostic analyses were performed by the Cox regression model. We found that nm23-H1-negative tumors were associated with a higher incidence of lymph-node metastasis (84.2%) than nm23-H1-positive ones (32.8%, p < 0.01). The distant metastasis and loco-regional recurrence rates in the nm23-H1-negative group were 55.8% and 31.68%, respectively but only 17.2% and 11.5%, respectively, in the nm23-H1-positive group (p < 0.01). A significant association was found between expression of nm23-H1 protein and prognosis (p < 0.01). Expression of nm23-H1 protein indicated favorable prognosis, suggesting that the absence of nm23-H1 protein expression was significantly associated with lymph-node metastasis, recurrence and distant metastasis in NPC. Expression of the nm23-H1 gene may be valuable for assessing the prognosis of NPC. Int. J. Cancer (Pred. Oncol.) 79:596–600, 1998. © 1998 Wiley-Liss, Inc.     

Correlation between nm23-H1 overexpression and clinicopathological variables in human anal canal carcinoma.

To improve life expectancy prognostic factors other than TNM have been investigated. It is thought that nm23 protein may play a specific biological role in suppressing tumor metastasis. The purpose of this study was to elucidate the clinical significance of nm23 expression in human anal canal carcinoma. Immunostaining using anti-nm23 monoclonal antibody was performed in 22 anal canal tumors. The results were correlated with clinicopathological variables. Six cases out of 22 (27.3%) were nm23-positive. Significant association was found between nm23-H1 expression and depth of invasion, lymph node involvement and prognosis (p<0.05). There was no significant association between nm23-H1 expression, histologic type and age of the patients. nm23-H1 expression was not seen in our cases with metastasis and this may be related to nm23 gene alterations not being detectable by the monoclonal antibody used or to the presence of a subset of tumors in which nm23 gene abnormalities had not yet occurred at the time of tumor excision or biopsy. Overexpression of nm23-H1 protein in anal canal carcinoma may have implications for its metastatic potential. nm23-H1 expression would provide a more accurate evaluation of outcome for individual patients and thus improve treatment planning.     

Expression of nm23-H1 in uveal melanoma

Uveal melanoma (UM) is the most common malignant intraocular tumor in adults. Despite the high accuracy of clinical diagnosis and advances in local treatment, more than 50% of UM patients develop metastasis within 10 years of initial diagnosis. NM23 is one of the human metastasis suppressor genes. Reduced nm23-H1 expression is correlated with high metastatic potential in many different cancers. The purpose of this study is to investigate the expression of nm23-H1 in UM and its potential value as a prognostic marker. Immunostaining of nm23-H1 was verified in five human UM cell lines with different metastatic potentials. The expression level of nm23-H1 mRNA was evaluated with one-step quantitative real-time PCR. The invasion ability of the cell lines was assessed before and after silencing nm23-H1 with small interference RNA. Thirty-two cases of paraffin-embedded specimens of human UM were immunostained with nm23-H1 monoclonal antibody. The immunostaining was evaluated in a semiquantitative fashion based on extent and intensity. The real-time PCR results of five human UM cell lines showed that expression of nm23-H1 was higher in cell lines with low metastatic potential compared with those with high metastatic potential (P<0.05). The invasive ability of the UM cell lines increased after silencing nm23-H1 expression with small interference RNA (P<0.05). The immunostaining of nm23-H1 was cytoplasmic in all cell lines and UM patients samples. The increased immunostaining intensity of nm23-H1 in patients' samples was associated with better survival rate (Kaplan-Meier test P=0.0097). The expression of nm23-H1 was not correlated with other prognostic factors. It can be concluded that nm23-H1 may be a prognostic marker to predict the survival rate of UM patients and it has the potential to identify high-risk patients.     

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.     

食管鳞状细胞癌p16和nm23-H1基因蛋白表达及意义

目的 :探讨多肿瘤抑制基因p16蛋白和nm2 3 H1基因蛋白在人食管癌的表达及与肿瘤生物学行为的关系。方法 :应用免疫组织化学方法观察 51例食管鳞状细胞癌及其淋巴结转移癌中p16蛋白的表达 ,结合观察转移抑制基因nm 2 3 H1的表达。结果 :p16在食管鳞状细胞癌中呈低表达 ( 4 5 1% ,2 3/51) ,阳性率显著低于正常食管粘膜及癌旁组织 (P <0 0 1) ,阳性率与分化程度相关 (P <0 0 1) ,Ⅰ级66 7% ( 14/ 2 1) ,Ⅱ级 34 8% ( 8/ 2 5) ,Ⅲ级 14 3% ( 1/ 7) ;与肿瘤浸润、转移无关。nm 2 3 H1的阳性率为60 8% ( 31/ 51) ,与浸润深度呈负相关 (P <0 0 5) ,淋巴结转移癌的阳性率 ( 18 8% ,3/ 16)显著低于食管癌原发灶 (P <0 0 1)。p16与nm 2 3 H1的表达无关。结论 :p16蛋白在食管癌的低表达提示有频发性的p16基因失活 ,且与肿瘤分化有关。nm2 3 H1与食管癌的浸润 ,转移相关。p16和nm 2 3 H1的失活在食管癌的形成及发展中可能起不同的作用