Expression of nm23 gene in hepatocellular carcinoma tissue and its relation with metastasis

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NORTHERN BLOT ANALYSIS OF nm23 GENE EXPRESSION IN HUMAN LUNG CANCER

Objective: To investigate the role of nm23 gene expression in human lung cancer. Methods: Forty human lung cancer tissues and 19 non-cancer pulmonary tissues were studied for their nm23-H1 and nm23-H2 mRNA expression with non-radioactive Northern blot hybridization. The correlation of nm23 mRNA expression with clinical features of lung cancer was analyzed. Results: The mRNA expression of nm23-H2 gene in poorly differentiated squamous cell carcinoma was significantly decreased compared to that in moderate-high differentiated squamousd cell carcinoma. The mRNA expression of nm23-H1 and nm23-H2 gene in small cell lung cancer was significantly decreased compared to that in squamous cell carcinoma. No significant difference in nm23 mRNA expression was observed between lung cancer with and without lymph node metastasis, nor was there significant difference between tumor stage. Conclusion: The mRNA expression of nm23 gene is correlated with the degree of differentiation of lung cancer, but there is no evidence of metastasis suppression effect by nm23 gene.

EXPRESSION OF NM23-H1 GENE PRODUCT IN NASOPHARYNGEAL CARCINOMA AND ITS CLINICAL SIGNIFICANCE

Objective: The nm23 gene is one of the tumor metastatic suppressor genes. The expression of nm23H1 has been reported to be inversely associated with metastatic potentiality in a number of human carcinomas, including breast, colorectal, gastric, hepatocellular and gallbladder carcinomas. In this study, the immunohistochemical staining of nm23H1 protein in human nasopharyngeal carcinoma (NPC) was examined, and the relationship between nm23H1 and both metastasis and prognosis of patients with NPC was also investigated. Methods: Routine LSAB immunohistochemistry with the nm23H1 monoclonal murine antibody was employed to study the expression of nm23H1 protein in 95 paraffinembedded specimens of NPC treated at our hospital. The clinical pathologic data and results of followup were also retrieved. Comparisons between patients with and without expression of nm23H1 protein with respect to metastasis, locoregional recurrence and survival were performed using Log rank test. Multivariate prognostic analyses were performed by using Cox's regression model. Results: Nm23H1 negative expressive tumors were associated with a higher incidence of lymphnode metastasis (86.7%) than those of nm23H1 positive (48.6%, P<0.01). Nm23H1 negative expressive tumors were associated with a high incidence of recurrence and distant metastasis after radiotherapy (P<0.05). A significant association was found between expression of nm23H1 and prognosis (P<0.01). The expression of nm23H1 indicated favorable prognosis. Conclusion: It was suggested that nm23 H1 negative expression was significantly associated with lymphnode metastasis, recurrence and distant metastasis. Nm23H1 may have value for predicting the prognosis of NPC.

EVALUATION OF THE PROGNOSTIC VALUE OF nm23 GENE EXPRESSION IN BREAST CANCER

Objective: To investigate the expression of nm23 gene and evaluate its prognostic value in breast cancer. Methods: nm23 expressions were detected in 101 breast cancer patients (group 1) by immunohistochemistry. RT-PCR and immunohistochemistry were used to measure expressions of nm23 gene in another 68 patients with breast cancer (group 2). Results: nm23 gene expression in group 1 was inversely associated with distant metastasis and lymph node metastasis (P<0.05). In 44 patients with negative lymph node, 9 cases progressed to distant metastasis, 7 of them (77.8%) showed low expression of nm23 gene (P<0.05). In 57 patients with positive lymph node, 24 our of 29 patients who had no distant metastasis (82.8%) expressed nm23 gene at high level (P<0.05). Meanwhile, there were 6 patients with distant metastasis in the group 2, all of thenm expressed nm23 gene mRNA at low level. Conclusion: The results showed that nm23 gene might play an independent role in predicting prognosis of breast cancer.

THE HIGH-LEVEL EXPRESSION OF nm23(NDP)GENE IN NPC

We have studied the expression of nm23(NDP) in 50 cases nasopharyngeal biopsies with anti-nm23(NDP) antibodies. As a result, the NDP positive rate in nasopharyngeal carcinoma (NPC) (95.54%) markedly increased (P<0.05), as compared with that in the normal nasopharyngeal epithelia (50.00-60.00%) and lymphocytes (52.00%). There were cytopfasmic type, nucleus type and mixed cytoplasmonucleus type according to NDP location in a cell. Their positive rates were 64.44%, 15.56% and 20.00% respectively in nasopharyngeal carcinoma. The expression of NDP had no relation with cervical lymphometastases in NPC, and the NDP positive rates had no significance between bilateral cervical lymphometastases and unilateral (P<0.05). But the NDP expression had most relation with the NPC staging. The expression rate and the intensity in Ⅲ or Ⅳ stage patients were markedly higher than that in II stage. It points out that the high-level expression of NDP had relation with the rapid cellular proliferation in NPC, and it may indicate the bad prognoses.

Expression of the novel gene NM23-H1B in ovarian cancer

Objective:To study the expression of the human novel gene NM23-H1B in ovarian cancer. Methods: Totally 24 samples from patients with epithelial ovarian tumor at different clinical stages and 4 from normal ovaries were examined for NM23-H1B mRNA expression by RT-PCR and Northern blot. Results: All samples expressed NM23-H1B mRNA through RT-PCR, while the level of expression in ovarian tumor was higher than that of normal ovary. The results of Northern blot showed that NM23-H1B was overexpressed in ovarian cancer while lowexpressed in normal ovary or low malignant potential (LMP). The level of expression at early stage cancer(stageⅠand Ⅱ) was higher than those in advanced cancer(stage Ⅲ and Ⅳ). In early stage carcinoma, the expression level was involved in the differentiation of tumor cell, and well-differentiated cancer expressed NM23-H1B mRNA in comparatively higher level. Conclusion: The novel gene NM23-N1B is closely correlated with the ovarian cancer.

IDENTIFICATION OF A NEW HUMAN nm23 GENE,nm23-H3b

Nm23 is a kind of effective tumor metastasis suppressor genes which included two types in human:nm23-H1 and nm23-H2. Amino acid identity between nm23-H1 and nm23-H2 was 88%.In this study,using a pair of primers to flank the part of coding sequence of nm 23,the 5'-translated sequence was amplified by polymerase chain reaction (PCR) from human normal liver genomic DNA.A 375-base pairs clone was charactertzed,which designated pnm23-H3b.The nm23-H3b nucleotide sequence between 40 bp and 70 bp is different from nm23-H1 and nm23-H2,and other sequences have 86%and 90%identical to nm23-H1 and nm23-H2,respectively.Southern blot containing BglⅡ-digested human liver genomic DNA hybridized to the entire nm23-H3b DNA and showed three bands at 10.5,7.9 and 4.0 Kb.These data demonstrate that the third human nm23 exists possibly.Therefore,nm23 may be considered a family of closely related genes.

乳腺癌原发灶和淋巴结转移灶中VEGF、nm23的表达变化及意义

目的：观察乳腺癌原发灶及淋巴结转移灶中血管内皮生长因子（VEGF）及转移抑制基因（m23）的表达变化及意义。方法选择2005年1～12月在我院行手术治疗且术前均未行抗癌治疗的60例乳腺癌患者，术后病理证实无同侧腋窝淋巴结转移24例（无转移组），伴有同侧腋窝淋巴结转移36例（转移组），采用免疫组化SP法检测乳腺癌原发灶和淋巴结转移灶中VEGF、nm23的表达，并分析VEGF与nm23的相关性。结果转移组原发灶中VEGF阳性表达率为91．7％，转移灶中为61．1％，无转移组VEGF阳性表达率为70．8％，转移组原发灶中VEGF阳性表达率明显高于转移灶及无转移组（P均＜0．05）。转移组nm23阳性表达率在原发灶中为16．7％，转移灶中为41．7％，无转移组nm23阳性表达率为45．8％，转移组原发灶中nm23阳性表达率明显低于转移灶及无转移组（P均＜0．05）。原发灶VEGF与nm23表达呈负相关（r＝-0．415，P＜0．05）。原发灶VEGF、nm23表达与淋巴结转移灶中VEGF、nm23表达无相关性。结论乳腺癌原发灶中VEGF表达明显高于、nm23表达明显低于淋巴结转移灶， VEGF、nm23与乳腺癌淋巴结转移密切相关，二者在乳腺癌发生和淋巴结转移中起拮抗作用。

nm 23基因参与肿瘤转移抑制机制的研究

目的：探讨ｎｍ２３基因参与肿瘤转移抑制的机制。方法：采用膜片钳制技术全细胞记录方式，对具有不同ｎｍ２３基因表达的不同转移潜能的人胰腺癌ＪＦ３０５单克隆细胞亚系中钙电流特征进行观察。结果：人胰腺癌ＪＦ３０５单克隆细胞亚系中的钙电流以钙释放激活的钙内流方式为主。人胰腺癌ＪＦ３０５Ａ、ＪＦ３０５Ｃ细胞钙电流的大小有显著差异（Ｐ＜０．０１）。结论：钙离子参与了ｎｍ２３基因在人胰腺癌ＪＦ３０５细胞亚系的转移抑制机制。

胃癌组织中Ezrin和nm-23的表达及临床意义

目的探讨胃癌组织中Ezrin和nm-23蛋白的表达及其与临床和预后的关系。方法应用免疫组化Envi-sion法检测164例胃癌癌组织、164例癌旁组织中Ezrin和nm-23的表达情况,分析两者之间的相互关系。结果 Ezrin和nm-23在胃癌癌旁组织中均无异常表达,在胃癌组织中Ezrin和nm-23的表达率分别为48.2%(79/164)和35.4%(58/164)。Ezrin在胃癌中的表达与淋巴结转移及临床分期呈正相关,而与无病生存时间呈负相关(P<0.05);nm-23的表达与淋巴结转移、临床分期呈负相关,而与无病生存时间呈正相关(P<0.05);Ezrin和nm-23在胃癌中的表达与年龄、肿块大小、性别无相关性(P>0.05)。结论检测Ezrin和nm-23在胃癌中的表达有助于判断胃癌的转移潜能和预后。

人喉鳞癌肿瘤转移抑制基因nm23-H1的分子遗传学研究

目的 在DNA水平上 ,进一步证实nm2 3基因与喉鳞癌转移的关联及在喉鳞癌中的突变模式。方法 应用PCR体外扩增技术、PCR SSCP银染技术及DNA测序技术对 2 3例喉鳞癌及对照正常组织基因组nm2 3 H1基因进行了分子遗传学研究。结果 在部分喉鳞癌转移灶组织中存在nm2 3 H1基因点突变 ,3个突变位点分别位于第 82个密码子 (TTT CTT) ,第 10 6密码子 (GTT ATT)和第 12 1密码子 (ATG—GTG)。除第 10 6密码子为无意义突变外 ,其余二处突变编码的氨基酸分别由苯丙氨酸及蛋氨酸变成谷氨酸及组氨酸。结论 ①应用PCR SSCP银染技术检测喉癌nm2 3 H1转移抑制基因突变是一种快速、有效的实验手段。它具有敏感性高 ,不需特殊仪器和昂贵试剂 ,实验成本低廉 ,实验步骤简便等优点。②nm2 3 H1基因突变在喉鳞癌的转移中起一定的作用。一方面可能使微管聚合异常而引起减数分裂时纺锤体的异常 ,从而导致癌细胞染色体非整倍体的形成 ,进而促进肿瘤的发展 ;另一方面它可能通过影响细胞骨架而引起细胞运动 。

膀胱癌中nm23-H1基质金属蛋白酶及其抑制物的表达及其临床意义

目的nm23-H1是抑制肿瘤转移的代表性基因,在人类乳腺癌、肝癌、胃癌、卵巢癌、恶性黑色素瘤等肿瘤的研究中均发现nm23-H1表达下降或不表达与肿瘤的转移过程以及不良预后密切相关;基质金属蛋白酶-2(MMP-2)及其抑制物(TIMP-2)调控着细胞外基质的降解,在肿瘤浸润、转移过程中发挥重要作用。因此,探讨膀胱癌组织中nm23-H1、MMP-2与TIMP-2的蛋白表达及其与临床、病理指标之间的关系具有非常重要的意义。方法采用免疫组织化学SP法分析69例手术切除膀胱癌中nm23与MMP-2、TIMP-2蛋白的表达情况。结果nm23-H1、MMP-2在正常膀胱黏膜与膀胱移行细胞癌(TCCB)中的表达差异有统计学意义(P<0.01),其中nm23-H1表达与肿瘤病理分级、临床分期以及是否转移呈负相关,MMP-2蛋白表达与肿瘤分级、分期呈正相关;在肿瘤组织中随TCCB临床病理分期升高nm23-H1的表达呈现降低趋势,而MMP-2表达随TCCB临床病理分期升高呈现增高趋势,且差异有统计学意义(P<0.05)。此外,MMP-2在≥2 cm的肿瘤中有较高的表达率,且差异有统计学意义(P<0.05)。TIMP-2在TCCB中的表达随病理分级、临床分期的增高而降低,但差异无统计学意义(P>0.05)。结论检测膀胱癌转移相关基因表达并分析其表达的相互关系,有助于判断膀胱癌的转移潜能及指导术后干预治疗。

抑癌基因p16和转移抑制基因nm23在卵巢上皮性肿瘤中的表达

目的:探讨抑癌基因p16及转移抑制基因nm23与卵巢上皮性肿瘤病理特点及生物学行为的关系。方法:免疫组化SP法检测30例正常卵巢,50例卵巢上皮性良性肿瘤及50例卵巢癌组织中P16蛋白和nm23蛋白,并分析其阳性表达与肿瘤的关系。结果:nm23蛋白在卵巢癌组织中的阳性率(70.0%)明显高于正常卵巢(43.3%)及良性肿瘤组(44.0%)(P<0.05);在浆液性癌(87.5%)和粘液性癌(47.8%)中的阳性差率也有显著差异(P<0.05);特别是在有淋巴结转移(39.1%)和无淋巴结转移(92.6%)的中检出率相非常显著(P<0.01),而与卵巢癌的临床分期及病理分级无明显相关性。p16蛋白在卵巢癌中的检出率(26.0%)明显低于正常卵巢(80.0%)和良性上皮性肿瘤(52.0%)(P<0.01),在晚期肿瘤中的表达率(20.5%)也远低于早期肿瘤(63.6%),而且在高度恶性组织中表达水平(11.1%)与中低度恶性组织的表达水平(43.5%)也有显著差异(P<0.05)。结论:nm23基因的表达与卵巢癌的淋巴结转移等生物学行为有关,p16基因可作为临床判断预后的重要指标之一。nm23、p16基因与卵巢上皮性肿瘤的发生、发展密切相关。

P-选择素和nm_(23)基因蛋白在食管鳞癌中的表达及其与肿瘤转移关系的研究

目的 :研究P 选择素和nm2 3 基因蛋白在食管鳞癌中的表达及其意义。方法 :以手术切除食管癌标本做病理组织切片 ,采用免疫组织化学染色 ,对整张切片中血管和细胞核、细胞浆或细胞膜的着色情况进行观察 ,确定其阳性或阴性。结果 :①P 选择素、nm2 3 基因蛋白的表达与食管磷癌的组织学分级无关。②P 选择素表达随着肿瘤的增大及淋巴结转移而增强 ,Ⅲ、Ⅳ期癌高于Ⅰ、Ⅱ期癌。③nm2 3 基因表达与TNM分期无关 ,在淋巴结转移组中阳性率显著低于无淋巴结转移组。④P 选择素和nm2 3 基因在食管癌中的表达呈显著负相关关系。结论 :P 选择素高表达与nm2 3 基因低表达共同提示食管癌浸润转移的高风险性。

THE ASSOCIATION OF THE EXPRESSION OF MTA1, NM23H1 WITH THE INVASION, METASTASIS OF OVARIAN CARCINOMA

Objective. To understanding the molecular mechanisms in invasion andmetastasis of the ovarian carcinoma, we investigate a novel candidate metastasis―associated gene (MTA1) and nm23Hl mRNA expression and mutation in ovarian carcinoma. Methods. Twenty primary ovariancarcinoma specimens, 20 corresponding lymph nodes and 8 normal ovarian was examined for mRNAexpression and mutation of MTA1 and nm23Hl genes by reverse-transcription ploymerase chain reaction(RT―PCR) and RT―PCR―SSCP analysis. The level of the expression was determined by the relativeoptic desity (ROD) of the PCR products. Results. The frequency of MAT1 overexpression was 100% (7/7)in primary ovarian carcinoma with metastasis but only 38.5% (5/13) in those without metastasis(P=0.0103). Overexpression of MAT1 was observed in 87.5% (6/7) of lymph nodes with metastasis butonly 23% (3/13). of lymph nodes without metastasis (P=0.0118). In contrast with MAT1, low expressionof nm23H1 mRNA was seen in 7 of 7 o-varian carcinoma with metastasis but only in 4 of 13(30%) ofthose without metastassis (P=0.0043). Low nm23H1 expression was also seen in 7 of 7 lymph nodes withmetastasis but only in 5 of 13 (38.5%) nonmetastatic lymph nodes (P=0.0102). The ROD ratio of MAT1to nm23Hl increased with the development of metastasis. No mutation of MAT1 and nm23H1 genes wasfound by SSCP analysis. Conclusion. The mRNA expression of MTA1 and nm23H1 is positively andnegatively correlated with lymph node metastasis, respectively. Expression abnormalities but notmutation of the two genes are frequent events related to lymph node metastasis of ovarian cancer.

nm23 expression in gastric carcinoma and its relationship with lymphoproliferation

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NM23基因与肿瘤生长转移相互关系的探讨

NM23基因是人类较早发现的抑癌基因,NM23基因与恶性肿瘤生长,向周围组织浸润以及向远处组织器官的转移已经成为近年来研究的热点。通过对NM23基因与肿瘤生长转移之间相互关系的探讨有利于我们更好去认识NM23基因,也有利于我们对恶性肿瘤生长浸润转移的认识。本文主要对NM23基因以及它与肿瘤生长转移之间相互关系作一概述。

Relation of nm23 gene expression to CT sign and prognosis in peripheral nonsmall cell lung cancer

CT observations and the cellular factors of molecular biology each enable one to recognize macro/micro changes in lung cancer. Growing pattern, characteristic shapes, degree of malignancy, relapse and metastasis of lung cancer are mainly determined by their molecular biology. Change of tumour shape, determined by the tumour’s biological behaviour, is the basis of CT observations. That is, the pathological change acts as a bridge which links the CT observation to molecular biology and makes the investigation of internal relationship between CT sign and molecular biology behaviour possible. As a tumour suppressor gene, nm23 gene is located in chromosome 17q21.3, encoding a nucleoside diphosphate kinase.^(1,2) We studied the expression of nm23 in peripheral nonsmall cell lung cancer (NSCLC) using the streptavidin peroxidase (SP) immunohistochemical method and investigated retrospectively the relationship between nm23 gene, CT observation, biological behaviour and prognosis of NSCLC.

乳腺癌组织ezrin、nm-23和VEGF的表达及临床意义

目的探讨乳腺癌组织中膜-细胞骨架联接蛋白埃兹蛋白(ezrin)、肿瘤转移抑制因子(nm-23)和血管内皮生长因子(VEGF)等蛋白表达与乳腺癌临床因素及预后的关系。方法应用免疫组化S-P方法检测63例手术切除的乳腺癌组织标本中ezirn、nm-23、VEGF的表达,并与临床进行对比分析。结果ezirn、nm-23和VEGF阳性表达率分别为55.56%、52.38%和65.08%)。三项指标与淋巴结转移、临床分期及无病生存时间均密切相关(P<0.05),但与年龄、肿块大小及月经情况均无相关性。结论Ezirn、nm-23和VEGF在乳腺癌组织中表达较高,可能对其发展起促进作用,并可能作为评价预后的指标。

PCNA和C-erbB-2及nm-23-H1在乳腺癌组织中的表达及临床意义

目的探讨PCNA C-erbB-2和nm-23-H1在乳腺癌组织中的表达及其与病理分型、临床分期、淋巴结转移、肿瘤大小、组织学分级的关系。方法采用免疫组织化学SP法测定76例乳腺癌组织石蜡切片中PCNA C-erbB-2和nm-23-H1蛋白的表达。结果PCNA C-erbB-2在乳腺癌组织中的表达与肿瘤的组织学分级、淋巴结转移数目、TNM分期呈正相关(P<0．01),与病理分型、肿瘤大小无关。nm-23-H1与淋巴结转移数目、肿瘤大小、TNM分期呈负相关(P<0．01),与病理分型、组织学分级无关。PCNA和C-erbB-2的表达呈正相关(P<0．01)。C-erbB-2和nm-23-H1 的表达呈负相关(P<0．01)。PCNA和nm-23-H1的表达差异无统计学意义(P>0．05)。结论PC- NA C-erhB-2和nm-23-H1可以作为判断乳腺癌生物学行为,后续治疗和预后的三个重要的客观标记物。