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与肿瘤侵袭、转移的研究进展

肿瘤的侵袭和转移是多基因多因子共同作用的结果。据推测nm23-H1基因可能仅为转移相关基因的上游调节基因,它对肿瘤转移潜能的抑制作用是通过下游基因实现的。本文重点对nm23-H1的分子生物学、nm23-H1的相关因素及其与肿瘤侵袭转移的组织病理相关性研究进展进行综述。     

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基因表达与非小细胞肺癌转移的关系

目的探讨nm23-H1基因表达与非小细胞肺癌(NSCLC)转移的关系.方法应用免疫组织化学SABC方法对90例NSCLC组织标本进行nm23-H1表达的检测.结果 NSCLC的nm23-H1阳性表达率51.1%(46/90),和NSCLC不同组织学类型、病理分级、PTNM分期、性别、年龄及远处转移无关(P＞0.05);nm23-H1的阳性表达与淋巴结转移呈负相关(P＜0.01),与NSCLC患者术后生存期呈正相关(P＜0.05).结论 nm23-H1可作为预测NSCLC淋巴结转移及预后的指标.     

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.     

PTEN及nm23-H1蛋白表达在非小细胞肺癌转移中的意义

背景与目的PTEN及nm23-H1基因均被证实是肿瘤转移的抑制基因,目前大多数研究都停留在基础研究上,本研究通过检测PTEN及nm23-H1蛋白在非小细胞肺癌(NSCLC)中的表达,以探讨它们的临床意义及相互关系。方法应用免疫组织化学法检测60例非小细胞肺癌组织中PTEN及nm23-H1基因蛋白的表达。结果NSCLC无淋巴结转移组PTEN蛋白阳性表达率为79.31%,高于NSCLC伴有淋巴结转移组41.94%,两者差异有统计学意义(P<0.01);无淋巴结转移组nm23-H1蛋白阳性表达率为82.76%,高于NSCLC伴有淋巴结转移组45.16%,两者差异有统计学意义(P<0.01)。PTEN和nm23-H1蛋白表达在NSCLC中一致性较好(Kappa=0.4366,Z=3.3905,P<0.01),两基因可能有协同作用。结论PTEN及nm23-H1蛋白表达均与NSCLC的淋巴结转移有关,PTEN和nm23-H1蛋白均可作为预测肿瘤转移的重要指标,对两种蛋白进行免疫组织化学联合检测,可能更有利于肺癌预测淋巴结转移及预后。     

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.     

转移抑制基因nm23-H1参与肺癌Ras信号传导机理研究

背景与目的nm23-H1基因已被证实为一种肿瘤转移抑制基因,但其作用机理尚不明了,本研究探讨nm23-H1参与肺癌Ras信号传导的机理。方法应用脂质体法将pEGFP-nm23-H1野生型和突变型质粒(WT,H118F,S120G,P96S,S44A)转染人大细胞肺癌细胞株L9981,采用免疫共沉淀与Western blot方法检测野生型和突变型nm23-H1蛋白与Ras支架蛋白KSR的关系。结果在转染了野生型和突变型质粒的五个细胞株中,鼠抗nm23-H1免疫沉淀物在86KDa处可检测到KSR的阳性条带,而各组KSR的量应用单因素方差分析比较无显著性差异(F=0.190,P=0.938)。结论本研究结果表明nm23-H1与KSR存在相互作用,但这种相互作用与nm23-H1有无突变及突变位点无关,nm23-H1可能是通过KSR参与肺癌Ras信号传导的。     

肿瘤转移抑制基因nm23-H1对不同宫颈癌细胞侵袭和增殖的作用

背景与目的：nm23-H1是一种多效性基因,其抑癌作用有一定的组织特异性。本研究目的在于探讨nm23-HI对不同宫颈癌细胞增殖和侵袭的作用。方法：将真核表达载体pcDNA3．1-nm23-HI转染入宫颈癌细胞,Transwell小室法检测转染前后细胞侵袭性改变。M1Tr法绘制生长曲线,比较细胞增殖能力。流式细胞仪检测细胞周期改变。结果：与亲本细胞Caski、SiHa和空载体转染细胞Caski-3．1、SiHa-3．1相比．pcDNA3．1-nm23-H1转染细胞Caski—N和SiHa—N的侵袭力和增殖受到明显抑制（P〈0．05）,G2／M期和S期细胞比例降低（P〈0．05）,而G。／G．期细胞比例明显增加（P〈0．05）。nm23-H1对HeLa细胞增殖、侵袭及细胞周期均无明显影响（P〉0．05）。结论：nm23-H1对不同宫颈癌细胞的增殖和侵袭等细胞表型可产生细胞特异性的抑制作用。     

nm23-H1基因对肺癌细胞株恶性表型的逆转作用

目的:通过稳定、高效的基因转染方法将nm23-H1基因导入nm23-H1基因缺失的人肺大细胞癌细胞株L9981,探讨nm23-H1基因是否具有逆转肺癌恶性表型的功能。方法:利用阳离子脂质体介导的基因转染技术,将nm23-H1基因导入L9981细胞株;利用细胞生长曲线、克隆形成率、Boyden小室法等方法研究转染前后细胞体外增殖、黏附、侵袭、转移能力的改变。结果:转染后的L9981细胞株中有nm23-H1基因mRNA和蛋白的阳性表达;转染后细胞株体外增殖速度减慢,克隆形成率降低,黏附性以及侵袭性均降低。结论:nm23-H1基因对肺癌细胞株L9981的体外生长、侵袭、转移具有负调控作用,表明nm23-H1基因具有逆转肺大细胞癌恶性转移表型的能力,为应用nm23-H1基因治疗肺癌提供了客观依据。     

nm23-H1基因稳定转染肺癌细胞的鉴定

目的：通过稳定、高效、低毒的基因转染方法将nm23-H1基因导入该基因缺失的人肺大细胞癌株L9981,从基因及蛋白水平鉴定nm23-H1基因转染肺癌细胞的建立。方法：以JM109为宿主菌,转化质粒载体PCMV-neo-Bam-nm23-H1,制备nm23-H1基因的真核表达载体;利用阳离子脂质体Lipofectamine介导,将nm23-H1基因导入L9981细胞株。采用RT-PCR技术检测转染后nm23-H1基因的表达,并用Western blot技术检测转染后nm23-H1蛋白的表达。结果：转染后的L9981细胞株中可检测到nm23-H1 mRNA及nm23-H1蛋白的阳性表达。结论：建立表达nm23-H1基因的肺癌细胞株L9981,为研究nm23-H1对肺癌恶性转移表型的逆转作用作基础准备。     

nm23-H1基因对肺癌细胞株恶性表型的逆转作用

目的：通过稳定、高效的基因转染方法将nm23-H1基因导入nm23-H1基因缺失的人肺大细胞癌细胞株L9981,探讨nm23-H1基因是否具有逆转肺癌恶性表型的功能。方法：利用阳离子脂质体介导的基因转染技术,将nm23-H1基因导入L9981细胞株;利用细胞生长曲线、克隆形成率、Boyden小室法等方法研究转染前后细胞体外增殖、黏附、侵袭、转移能力的改变。结果：转染后的L9981细胞株中有nm23-H1基因mRNA和蛋白的阳性表达;转染后细胞株体外增殖速度减慢,克隆形成率降低,黏附性以及侵袭性均降低。结论：nm23-H1基因对肺癌细胞株L9981的体外生长、侵袭、转移具有负调控作用,表明nm23-H1基因具有逆转肺大细胞癌恶性转移表型的能力,为应用nm23-H1基因治疗肺癌提供了客观依据。     

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 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.     

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     

The N-myc and c-myc downstream pathways include the chromosome 17q genes nm23-H1 and nm23-H2

Gain of chromosome 17q material is the most frequent genetic abnormality in neuroblastomas. The common region of gain is at least 375 cR large, which has precluded the identification of genes with a role in neuroblastoma pathogenesis. Neuroblastoma also frequently show amplification of the N-myc oncogene, which correlates closely with 17q gain. Both events are strong predictors of unfavorable prognosis. To identify genes that are part of the N-myc downstream pathway, we constructed SAGE libraries of an N-myc transfected and a control cell line. This identified the chromosome 17q genes nm23-H1 and nm23-H2 as being 6-10 times induced in the N-myc expressing cells. Northern and Western blot analysis confirmed this up-regulation. Time-course experiment shows that both genes are induced within 4 h after N-myc is switched on. Furthermore, we demonstrate also that c-myc can up-regulate nm23-H1 and nm23-H2 expression. Neuroblastoma tumor and cell line panels reveal a striking correlation between N-myc amplification and mRNA and protein expression of both nm23 genes. We show that the nm23 genes are located at the edge of the common region of chromosome 17q gain previously described in neuroblastoma cell lines. Our findings suggest that nm23-H1 and nm23-H2 expression is increased by 17q gain in neuroblastoma and can be further up-regulated by myc overexpression. These observations suggest a major role for nm23-H1 and nm23-H2 in tumorigenesis of unfavorable neuroblastomas.     

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。     

人类nm23-H1基因重组腺病毒的构建及鉴定

目的构建人肿瘤转移抑制基因nm23-H1的重组腺病毒载体。方法通过PCR方法以pcDNA3．1-nm23-H1为模板扩增出编码152个氨基酸的nm23-H1基因,并连接入穿梭质粒 pShuttle-CMV。将含有目的基因的重组穿梭质粒pShuttleCMV-nm23-H1经Pme I线性化后转化入 AdEasier-1感受态细胞,用含硫酸卡那霉素的LB培养基筛选重组子,并用PCR及酶切方法鉴定。鉴定正确的pAd-nm23-H1质粒经Pac I线性化后转染293细胞,包装重组腺病毒Adeno-nm23- H1,并进行PCR鉴定及病毒滴度测定。结果 PCR、酶切鉴定及测序结果证实pShuttleCMV-nm23- H1及pAd-nm23-H1质粒正确构建,收获病毒后的PCR鉴定及DNA测序结果证明Adeno-nm23- H1包被成功且无野生型腺病毒产生。重组腺病毒Adeno-nm23-H1滴度为4．3×109／ml。结论成功构建了重组腺病毒载体Adeno-nm23-H1,为进一步研究nm23-H1抑制肿瘤转移分子机制及基因治疗奠定了基础。     

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.     

人类nm23-H1基因重组腺病毒的构建及鉴定

目的构建人肿瘤转移抑制基因nm23-H1的重组腺病毒载体.方法通过PCR方法以pcDNA3.1-nm23-H1为模板扩增出编码152个氨基酸的nm23-H1基因,并连接入穿梭质粒pShuttle-CMV.将含有目的基因的重组穿梭质粒pShuttleCMV-nm23-H1经Pme Ⅰ线性化后转化入AdEasier-1感受态细胞,用含硫酸卡那霉素的LB培养基筛选重组子,并用PCR及酶切方法鉴定.鉴定正确的pAd-nm23-H1质粒经Pac Ⅰ线性化后转染293细胞,包装重组腺病毒Adeno-nm23-H1,并进行PCR鉴定及病毒滴度测定.结果PCR、酶切鉴定及测序结果证实pShuttleCMV-nm23-H1及pAd-nm23-H1质粒正确构建,收获病毒后的PCR鉴定及DNA测序结果证明Adeno-nm23-H1包被成功且无野生型腺病毒产生.重组腺病毒Adeno-nm23-H1滴度为4.3×109/ml.结论成功构建了重组腺病毒载体Adeno-nm23-H1,为进一步研究nm23-H1抑制肿瘤转移分子机制及基因治疗奠定了基础.