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.     

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和p53基因在乳腺癌细胞增殖及转移评估中的价值研究

目的探讨乳腺癌中nm23-H1、p53基因与肿瘤细胞增殖活性、淋巴结转移之间的关系。方法应用免疫组化技术检测乳腺癌组织中nm23-H1、p53基因蛋白及增殖指标k i-67的表达,结合临床及病理指标,探讨其临床意义。结果乳腺癌组织中nm23-H1、p53蛋白阳性表达率分别为50.8%(61/120)和60.0%(72/120),nm23-H1蛋白的表达与增殖程度(k i-67)和腋窝淋巴结转移有关,k i-67低增殖组nm23-H1阳性表达率为68.0%(49/72),而k i-67高增殖组nm23-H1阳性表达率则为25.0%(12/48),差异有显著性(P<0.05)。无合并淋巴结转移组中nm23-H1阳性表达率为72.2%(39/54),明显高于淋巴结转移组的阳性率33.3%(22/66)(P<0.05)。p53蛋白表达与雌激素受体有关,雌激素受体阳性组p53阳性阳性表达率为55.8%(47/86),阴性组阳性表达率为73.5%(25/34),差异有显著性(P<0.05)。结论nm23-H1和p53基因均与乳腺癌生物学行为有关,检测它们在乳腺癌中的表达可用来评估肿瘤是否具转移和侵袭能力。     

宫颈癌中nm23-H_1基因的不同表达与淋巴结转移及预后关系研究

目的　探讨nm2 3 -H1基因在宫颈癌组织中表达及其意义。方法　采用免疫组化法检测 88例宫颈癌组织中nm2 3 -H1基因表达 ;对nm2 3 -H1表达与临床病理因素及预后关系进行分析。结果　宫颈鳞癌与腺癌的表达阳性率分别为 46 4% (2 6 /5 6 )和5 6 2 % (18/32 ) ;在与临床病理指标的相关性分析中发现 :宫颈腺癌中 ,无淋巴结转移者nm2 3 -H1阳性率明显高于有淋巴结转移者(P <0 0 1) ;而鳞癌中nm2 3 -H1的表达与淋巴结转移无关 (P >0 0 5 )。另外 ,nm2 3 -H1的表达与宫颈癌的临床分期、病理分级、病灶大小无关。回顾性预后调查发现 ,nm2 3 -H1阳性患者的生存率明显高于阴性表达者 ,在鳞癌及腺癌中 ,P <0 0 5和P <0 0 1。结论　nm2 3 -H1表达与宫颈腺癌淋巴结转移呈负相关 ;与鳞癌淋巴结转移无关。nm2 3 -H1基因的表达可以作为宫颈癌预后的一项指标     

NM-23 H1 immunohistochemistry is not useful as predictor of metastatic potential of colorectal cancer.

This study aimed to investigate whether immunohistochemical staining for nm23-H1 protein in the primary tumour is correlated with tumour stage, tumour differentiation, DNA ploidy, cell proliferative index, p53 status and patient survival time in colorectal cancer. Full-cross colorectal cancer biopsies were collected from 202 consecutive surgical specimens between 1987 and 1990. Immunohistochemical expression of nm23-H1 protein was investigated in cryosections, using a monoclonal anti-nm23-H1 antibody (clone NM 301). The staining pattern was classified as follows: strong homogeneous intensity, moderate homogeneous intensity, moderate focal intensity, or as negative. Immunohistochemical expression of p53 was investigated using a monoclonal anti-p53 antibody (DO-7). The DNA ploidy and cell proliferative index were determined by flow cytometry. Possible correlation between nm23-H1 staining patterns and the other studied tumour characteristics was explored at the end of 1994. Median survival time of living patients was 66 months, range 50-93 months. No correlation was found between various nm23-H1 staining patterns and tumour stage, cell proliferative index or p53 status. Nm23-H1-negative tumours and tumours with moderate focal staining intensity were less differentiated than tumours with strong homogeneous or moderate homogeneous staining intensity (P < 0.05). Of the nm23-H1-negative tumours, a significantly higher number was near-diploid rather than aneuploid, as compared with those expressing positive nm23-H1 (P < 0.05). The number of dead patients in Dukes'' stages B and C did not correlate significantly with the nm23-H1 staining pattern. The nm23-H1 staining pattern alone, or combined with either of the other explored tumour characteristics, did not correlate with patient survival time. Immunohistochemical studies of the nm23-H1 protein expression are of minor value in the staging and prognostic prediction of colorectal cancer.     

MTA1和nm23-H1蛋白表达与乳腺癌生物学行为之间的关系

目的探讨肿瘤转移相关基因（NTA1）和nm23-H1蛋白表达与乳腺癌生物学行为之间的关系。方法运用免疫组化S-P法检测56例乳腺癌组织中NTAl、nm23-H1蛋白的表达。结果56例乳腺癌组织中39例NTA1蛋白阳性表达,阳性率为69．6％,NTA1高表达与乳腺癌组织学分级、临床分期和淋巴结转移关系密切（P〈0．05）;3l例nm23-H1蛋白阳性表达,阳性率为55．4％,nm23-H1低表达与乳腺癌组织学分级、临床分期和淋巴结转移关系密切（P〈0．05）;乳腺癌组织中NTA1、nm23-H1蛋白表达呈负相关（P〈0．05）。结论NTA1和nm23-H1蛋白表达与乳腺癌分化程度、淋巴结转移和预后关系密切,可作为诊断乳腺癌患者转移复发的参考指标,并有望成为乳腺癌基因治疗的新靶点。     

Nm23-H1 suppresses tumor cell motility by down-regulating the lysophosphatidic acid receptor EDG2

Exogenous overexpression of the metastasis suppressor gene Nm23-H1 reduces the metastatic potential of multiple types of cancer cells and suppresses in vitro tumor cell motility and invasion. Mutational analysis of Nm23-H1 revealed that substitution mutants P96S and S120G did not inhibit motility and invasion. To elucidate the molecular mechanism of Nm23-H1 motility suppression, expression microarray analysis of an MDA-MB-435 cancer cell line overexpressing wild-type Nm23-H1 was done and cross-compared with expression profiles from lines expressing the P96S and S120G mutants. Nine genes, MET, PTN, SMO, FZD1, L1CAM, MMP2, NETO2, CTGF, and EDG2, were down-regulated by wild-type but not by mutant Nm23-H1 expression. Reduced expression of these genes coincident with elevated Nm23-H1 expression was observed in human breast tumor cohorts, a panel of breast carcinoma cell lines, and hepatocellular carcinomas from control versus Nm23-M1 knockout mice. The functional significance of the down-regulated genes was assessed by transfection and in vitro motility assays. Only EDG2 overexpression significantly restored motility to Nm23-H1-suppressed cancer cells, enhancing motility by 60-fold in these cells. In addition, silencing EDG2 expression with small interfering RNA reduced the motile phenotype of metastatic breast cancer cells. These data suggest that Nm23-H1 suppresses metastasis, at least in part, through down-regulation of EDG2 expression.     

Correlation study of Bcl-2, B7-H1, EGFR,VEGF and colorectal cancer

Objective: To analyze the expressions of Bcl-2, B7-H1, EGFR and VEGF in colorectal cancer for the further investigation of their correlations with the clinical pathological features of colorectal cancer. Method: Fresh colorectal cancer tissues and the expressions of Bcl-2, B7-H1, VEGF and EGFR in paraneoplastic normal mucosal tissues of 57 cases were tested by immunohistochemisty method, and the results were analyzed by SPSS10.0. Results: 1. Compared with paraneoplastic normal tissues, the expressions of Bcl-2 and B7-H1 in colorectal cancer tissues increased significantly with significant difference (P<0.05), while the expression of EGFR and VEGF in colorectal cancer tissues showed no significant difference with those in paraneoplastic normal tissue (P>0.05); 2. The correlation with clinical pathological features: there was significant difference of expression rates of EGFR between different genders (P<0.05); the expressions of BCL-2 and B7-H1 in colorectal cancer of the high- and medium- differentiated groups were significantly higher than those of the low-differentiated group, and the difference was significant (P<0.01); compared with the colorectal cancer patients without lymph node metastasis (Dukes stage A+B), the expression of B7-H1 in patients with lymph node metastasis (Dukes stage C+D) was significantly higher (P<0.05); 3. Within the high- and medium- differentiated colorectal cancer tissues, Bcl-2 expression rate in B7-H1 negative group was higher than the positive group with significant difference (P<0.01). Conclusions: In colorectal carcinoma, Bcl-2, B7-H1, EGFR and VEGF were all expressed, independent from age and depth of invasion. However, the expression level of Bcl-2 and B7-H1 correlated with tissue differentiation, and the latter also had correlation with tumor staging. Meanwhile, the short-term follow-up showed that high expression of Bcl-2/B7-H1 existed in death cases. Therefore, the expression detection of Bcl-2, B7-H1 might provide a clear understanding of the biological behavior of colorectal cancer, and was important for the diagnosis, treatment and prognosis judgment of colorectal cancer.     

乳腺癌组织蛋白酶D、c-erbB-2蛋白、nm23蛋白表达与淋巴结转移的关系

目的探讨乳腺癌组织蛋白酶Ｄ（ＣａｔｈｅｐｓｉｎＤ）,ｃ－ｅｒｂＢ－２,ｎｍ２３的表达与淋巴结转移的关系。方法应用免疫组织化学Ｓ－Ｐ法,检测１０８例乳腺癌Ｃａｔｈ－Ｄ、ｃ－ｅｒｂＢ－２、ｎｍ２３的表达情况。结果Ｃａｔｈ－Ｄ、ｃ－ｅｒｂＢ－２、ｎｍ２３阳性表达率分别为５８．３％,５４．６％,４８．１％。Ｃａｔｈ－Ｄ,ｃ－ｅｒｂＢ－２阳性表达率与乳腺癌腋窝淋巴结转移之间呈正相关（Ｐ＜０．０５）,ｎｍ２３的阳性表达率与淋巴结转移之间呈非常显著负相关（Ｐ＜０．０１）。结论Ｃａｔｈ－Ｄ、ｃ－ｅｒｂＢ－２的高表达与ｎｍ２３的低表达可能协同促进乳腺癌的淋巴结转移     

Survivin、Bcl-2、nm23-H1的表达及其与鼻咽癌病理类型、临床分期的关系

目的：研究Survivin、Bcl-2及mn23-H1的表达及其与鼻咽癌（NPC）病理类型、临床分期之间的关系。方法：应用免疫组化分析32例NPCSurvivin、Bcl-2及nm23-H1的表达。结果：32例NPC组织中Sur-vivin、Bcl-2及nm23-HI的表达率分别为：68．7％（22／32）、71．9％（23／32）、56．3％（18／32）;晚期（Ⅲ、Ⅳ期）患者Survivin、Bcl-2阳性表达率分别为90％（18／20）、90％（18／20）明显高于早期（I、Ⅱ期）患者的表达率33．3％（4／12）、41．6％（5／12）,差异均有显著性（P〈0．05）;未分化癌患者Survivin、Bcl-2阳性表达率分别为100％（10／10）、100％（10／10）明显高于低分化鳞癌患者表达率54．5％（12／22）、59．1％（13／22）,差异均有显著性（P〈0．05）;有颈部淋巴转移的患者Survivin、Bcl-2阳性表达率分别为69．2％（18／26）、76．9％（20／26）高于无颈部淋巴转移患者表达率66．6％（4／6）、50％（3／6）,但均未见显著性（P〉0．05）;晚期（Ⅲ、Ⅳ期）患者nm23-H的阳性表达率40％（8／20）低于早期（I、Ⅱ期）患者的表达水平100％（12／12）,差异有显著性（P〈0．05）,未分化癌患者的阳性表达率（7／10,70％）高于低分化鳞癌患者的表达率54．5％（13／22）,但差异无显著性（P〉0．05）,无颈部淋巴转移患者nm23-HI的阳性表达率100％（6／6）高于有颈部淋巴转移患者表达水平46．1％（12／26）,差异有显著性（P〈0．05）。结论：鼻咽癌中存在着Survivin、Bel-2及nm23-H1的高表达;Survivin、Bel-2与鼻咽癌的临床分期、病理类型具有相关性,而与颈部淋巴转移无相关性,mn23—H1与鼻咽癌的临床分期、颈部淋巴转移具有相关性,但与病理类型无相关性。     

nm23蛋白与C—erbB—2蛋白在乳腺癌中的表达及临床意义

目的：探讨乳腺癌中抑制转移基因ｎｍ２３的表达及其与Ｃ－ｅｒｂＢ－２、ＥＲ、ＰＲ以及肿瘤组织学分级、淋巴结转移等临床病理特性的相互关系。方法：应用免疫组化ＳＰ法检测７０例乳腺癌中ｎｍ２３蛋白、ＣｅｒｂＢ－２蛋白、ＥＲ、ＰＲ的表达。结果：乳腺癌中ｎｍ２３蛋白阳性率为６２．８６％,ｎｍ２３蛋白的表达与组织学分级、细胞核分级、淋巴结转移以及临床分期均有相关性（Ｐ〈０．０１）,但与Ｃ－ｅｒｂＢ－２蛋白、ＥＲ     

肿瘤转移相关基因与周围型肺癌CT征象的关系研究

摘要目的：探讨VEGF和nm23-H1基因表达水平与周围型肺癌的病理生理学特征和CT征象之间的关系。方法：回顾性分析确诊的47例周围型肺癌患者CT表现．同时采用免疫组化方法(LSAB法)检测47例肺癌组织中的VEGF和nm23-H1表达水平。结果：1)VEGF、nm23-H1表达水平与原发肿瘤大小、淋巴结转移状态，pTNM分期均有密切关系(P＜0．05)。2)VEGF与nm23-H1表达水平呈负相关(P＜0.05)。3)VEGF基因的高表达和nm23-H1基因的低表达均与分叶征、纵隔淋巴结转移有关(P＜0．05)，且VEGF的高表达也与空洞征有关：二者表达水平与毛刺征、空泡征、胸膜凹陷征和支气管征均无关(P＞0．05)。4)深分叶征、空泡征和支气管征与肿瘤大小有关(P＜0．05)，而与毛刺征、胸膜凹陷征和纵隔淋巴结转移均无关(P＞0.05)。结论：VEGF过度表达和nm23-H1低表达均与周围型肺癌的分叶征、空洞坏死征和纵隔淋巴结转移有密切关系。     

CD44V6,nm23-H1表达与子宫内膜癌临床及病理的关系

目的研究子宫内膜癌CD44V6,nm23-H1基因的不同表达与其临床及病理各因素间的关系.方法应用免疫组织化学S-P法检测CD44V6与nm23-H1在55例子宫内膜癌中的表达,分析其与子宫内膜癌的临床及病理各参数的关系.结果(1)CD44V6的表达率与子宫内膜癌的分期,组织病理分级及癌组织肌层浸润深度之间无相关关系.在腺癌中其表达率低于鳞腺癌,在淋巴结转移(LNM)阳性的病例中其表达率低于LNM阴性者.(2)nm23-H1的阳性表达率与子宫内膜癌的分期及组织类型之间无相关关系,与组织病理分级及肌层浸润深度呈负相关.在LNM(+)者中其表达率显著低于LNM(-)者.(3)CD44V6与nm23-H1的阳性表达率之间无相关关系.结论(1)CD44V6在子宫内膜腺癌中的阳性表达率及过度表达率均显著低于鳞腺癌.(2)CD44V6的低表达可促进盆腔淋巴结转移.(3)nm23-H1的低表达与高组织病理分级,深肌层浸润及淋巴结转移有关.(4)CD44V6的阳性表达率与nm23-H1的阳性表达率之间无相关关系.     

Elevation of breast carcinoma Nm23-H1 metastasis suppressor gene expression and reduced motility by DNA methylation inhibition

We hypothesize that elevation of Nm23-H1 expression in micrometastatic breast cancer cells may inhibit their metastatic colonization and further invasion, and induce differentiation, thus resulting in a clinical benefit. The current study investigated the possible contribution of DNA methylation to the regulation of Nm23-H1 expression, based on the observation that two CpG islands are present in its promoter. 5-Aza-2'-deoxycytidine (5-Aza-CdR), a DNA methylation inhibitor, increased the Nm23-H1 expression of 5 of 11 human breast carcinoma cell lines in vitro, including 3 of 3 metastatically competent lines. Increased Nm23-H1 expression was accompanied by a reduction in motility in vitro, with minimal effect on proliferation. Both increased Nm23-H1 expression and decreased motility were observed using low (75 nM) concentrations of 5-Aza-CdR. Array analysis of MDA-MB-231 breast carcinoma cells treated with 5-Aza-CdR confirmed the elevation of nm23-H1 mRNA, whereas relatively few other genes exhibited altered expression. Bisulfite sequencing of the two CpG islands in a panel of cell lines and in 20 infiltrating ductal carcinomas revealed that one island (-3090 bp to -3922 bp) exhibited infrequent differential methylation. The data indicate that DNA methylation inhibitors can directly or indirectly cause both elevation of Nm23-H1 expression and decreased function in one aspect of metastasis, motility.     

大肠癌中nm23-H1的表达与淋巴结转移的关系

 目的　探讨大肠癌中nm23-H1的表达与淋巴转移的关系。方法　应用免疫组化方法研究96例大肠癌中nm23-H1蛋白的表达。结果　nm23-H1蛋白低表达与淋巴结或远处转移显著相关(P<0.05);nm23-H1蛋白低表达预测大肠癌转移的灵敏性为88.4%,特异性为79.3%。结论　检测nm23-H1蛋白可以预测大肠癌淋巴结或远处转移,从而可能成为临床治疗的判断依据。     

Overexpression of h-prune in breast cancer is correlated with advanced disease status.

PURPOSE: The h-prune gene is involved in cellular motility and metastasis formation in breast cancer through interacting with the nm23-H1 protein. The aim of this study was to better define the clinical and pathologic role of h-prune in breast cancer patients. EXPERIMENTAL DESIGN: Using immunohistochemistry, we assessed h-prune and nm23-H1 protein expression in two series of breast cancer patients: (i) in 2,109 cases with pathologic reports on primary tumors and (ii) in 412 cases with detailed clinical information. To assess the role of DNA amplification in gene activation, the h-prune copy number was evaluated by fluorescence in situ hybridization analysis in 1,016 breast cancer cases. RESULTS: In the patients tested (n = 2,463), 1,340 (54%) had an increased level of h-prune expression; a positive immunostaining for nm23-H1 was observed in 615 of 2,061 (30%) cases. Overexpression of h-prune was associated with multiple gene copy number at chromosome 1q21.3 in a very limited fraction of cases (68 of 1,016; 6.7%), strongly indicating that alternative pathways induce h-prune activation in breast cancer. Multivariate Cox regression analysis showed that neither h-prune overexpression nor decreased nm23-H1 immunostaining is independent prognostic factors. However, a significant association of h-prune overexpression with either advanced lymph node status (P = 0.017) or presence of distant metastases (P = 0.029) was observed. CONCLUSIONS: Although not significantly correlated with overall survival, positive h-prune immunostaining identifies subsets of breast cancer patients with higher tumor aggressiveness. Further investigations using larger collections of advanced breast cancer patients are required for assessing the predictive role of h-prune in breast cancer.     

Keratin 19 mRNA measurement to detect micrometastases in lymph nodes in breast cancer patients.

We have used polymerase chain reaction (PCR) to measure keratin 19 mRNA in order to detect breast cancer cells invading axillary lymph nodes. In a consecutive series of 125 patients with primary breast cancer, 75 patients had no evidence of lymph node involvement by conventional histology. A total of 530 lymph nodes from these patients were examined and 106 (20%) gave a keratin 19 product detectable by Southern hybridisation. This correlated with primary tumour size (P<0.001). These 106 nodes came from 23 patients. Thus, using this technique, 23/75 (30.6%) patients were found to have evidence of lymph node involvement who would otherwise have been designated lymph node negative.     

nm23—H1基因与乳腺癌转移和预后的关系及其突变的研究

采用免疫组化染色分析100例乳腺癌组织的nm23－H1基因表达与腋窝淋巴结转移和预后的关系，发现阳性表达者的5年无瘤生存率为84．8％（39／46），明显高于阴性表达者的52．9％（18／34），P＜0．01。进一步对20例采用RNA提取、PT－PCR、PCR－SSCP等方法检测乳腺癌组织中nm23－H1基因mRNA表达和突变情况，发现有突变的乳腺癌其基因表达均为阴性，胶窝淋巴结转移均为阳性（P＝0．036）。研究结果提示：nm23－H1基因突变可能易发生于晚期乳腺癌，同时检测nm23－H1基因mRNA表达水平和突变情况可作为判断乳腺癌（尤其是无淋巴结转移乳腺癌）的一项指标。