卵巢癌高频转移细胞模型中nm23-H1基因表达的相关性研究

目的筛选高频转移卵巢恶性肿瘤细胞,研究不同转移潜能的细胞和 nm23的相关性。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 North-ern-blot 方法测定各类肿瘤细胞 nm23 mRNA 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。测定各类细胞 nm23 mRNA 表达水平与肿瘤转移特性呈负相关。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。     

卵巢癌高频转移细胞模型中nm23-H1基因表达的相关性研究

目的 筛选高频转移卵巢恶性肿瘤细胞，研究不同转移潜能的细胞和nm23的相关性。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Northera-blot方法测定各类肿瘤细胞nm23 mRNA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。测定各类细胞nm23 mRNA表达水平与肿瘤转移特性呈负相关。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基因表达与鼻咽癌的早发转移

目的探讨nm23-H1基因产物的表达与鼻咽癌早发转移的关系.方法应用免疫组化S-P法对95例鼻咽癌组织蜡块进行nm23-H1检测.结果nm23-H1的阳性表达率为47.4%(45/95),早发转移组nm23-H1阳性率(26.7%)低于无转移组(60.0%),差异有统计学意义(P＜0.05).nm23-H1表达与临床分期无关.结论nm23-H1阴性表达与鼻咽癌早发转移有关,nm23-H1的表达水平可能是预测中晚期鼻咽癌远处转移的一个重要的生物学指标.     

nm23基因表达与乳腺癌远处转移及预后相关性的研究

目的 探讨nm23基因表达与乳腺癌远处转移的关系及其预后意义。方法 选取两组共169例乳腺癌患者,应用免疫组化和RT-PCR方法检测nm23基因表达。结果 nm23基因表达与乳腺癌远处转移及腋淋巴结转移呈显著负相关,与生存率呈正相关,第1组病例发生远处转移的9例腋淋巴结阴性患者中,7例为nm23蛋白低表达;而29例未发生远处转移的腋淋巴结阳性患者中,nm23蛋白中表达及高表达者为24例（82.8%),第2组发生复发和远处转移的6例患者其nm23mRNA均低表达,nm23基因表达与腋淋巴结转移,肿瘤大小均为影响预后的因素,结论 nm23基因作为一个独立的预后指标应用,厅弥补常规指标的不足,具有重要的临床应用价值。     

nm23-H1与大肠癌淋巴结转移及预后的关系

目的探讨nm23-H1与大肠癌淋巴结转移及预后的关系.方法对280例大肠癌存档蜡块进行重新切片,采用nm23-H1单克隆抗体进行免疫组化染色(S-P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;nm23-H1蛋白表达阴性126例中75例有淋巴结的转移,表达阳性154例中36例有淋巴结的转移(P＜0.01).全组表达阴性者在DukesC、D期较A、B期多见(P＜0.01),随访108例中有61例发生肝转移;表达阳性者随访167例中有41例发生肝转移(P＜0.01),有5例失访者.nm23-H1蛋白表达阴性、阳性、强阳性组的五年累积生存率分别为39.1%(44/129)、58.3%(42/72)、68.3%(51/79)(P＜0.01).结论 nm23-H1蛋白表达与大肠癌淋巴结转移、肝转移呈负相关.检测大肠癌组织切片中的nm23-H1蛋白表达,可能成为预测大肠癌淋巴结转移及预后的生物学指标之一.     

nm23-H1表达预测大肠癌转移的价值研究

 目的　大肠癌预后取决于是否存在肿瘤转移,目前尚无肯定的观测大肠癌细胞恶性潜能的生物学指标。本研究目的在于探讨nm23-H1作为预测大肠癌转移指标的潜在价值。方法　采用免疫组织化学链霉菌亲合物素蛋白—生物素酶标(SP)方法对58例存档大肠癌标本进行nm23-H1蛋白表达检测,用SSPS统计软件包对nm-H1表达与临床病理参数的关系进行分析。结果　正常大肠粘膜和大肠腺瘤nm23呈阳性表达,62.1%大肠癌组织阳性表达。nm23表达与肿瘤分级、淋巴结及肝转移负相关(P值分别为0.0100,0.2087,0.00376)。nm23阳性表达的大肠癌患者预后好(P=0.0002)。结论　nm23-H1是预测大肠癌细胞转移潜能和预后的敏感指标。     

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。     

P53 is a regulator of the metastasis suppressor gene Nm23-H1

p53, a tumor suppressor gene involved in the G1 cell cycle checkpoint, is also the most frequently mutated gene in human cancer. In addition, p53 modifies the ability of tumor cells to metastasize. The metastasis-associated gene Nm23-H1, which encodes an 18-kDa nucleoside diphosphate kinase, was previously identified in cells with low metastatic potential. Although p53 and Nm23-H1 proteins play an important part in regulating the progression of cancer, any functional relationship between these two proteins is currently unknown. Here we report an association between p53 levels and expression of the Nm23-H1 gene. Our data indicate that wild-type (wt) p53 upregulated the expression of Nm23-H1 at protein and mRNA levels in MCF-7 and J7B cells. This capacity of wt p53 to regulate expression of Nm23-H1 was not only dependent on the endogenous but also the exogenous origin of p53, and could not be reproduced with mutant p53. Subsequently, the invasive ability of MCF-7 and J7B cells was suppressed upon induction of the Nm23-H1 protein by p53. In contrast, increased levels of p53 downregulated the expression of Nm23-H1 at the protein and mRNA levels in RKO and H1299 cells and, as a consequence, increased the invasive ability of both cell types. Thus, our results implicated the differential regulation of Nm23-H1 by p53 in different cell types as an important component in the molecular mechanisms of tumor metastasis.     

Nm23-H1 expression of metastatic tumors in the lymph nodes is a prognostic indicator of oral squamous cell carcinoma

We recently reported that low Nm23-H1 expression of primary oral squamous cell carcinoma (OSCC) was correlated with the occurrence of lymphatic metastasis. However, little is known about whether Nm23-H1 level of metastatic tumors in the cervical lymph nodes is reduced in comparison with primary oral cancers and its significance for patients' prognosis. By immunohistochemistry, we analyzed the Nm23-H1 expression in 52 pairs of OSCC specimens from primary oral cancers and their metastatic lymph nodes. Western blot analysis further confirmed the immunohistochemical interpretation. To verify the effects of Nm23-H1 on cell migration and invasion, we established several stable clones derived from a human OSCC cell line (SAS) by knockdown and overexpression. Wound-healing closure, transwell migration and invasion assays were performed to determine cell motility, migratory and invasive activities. Western blot analysis was carried out to evaluate cyclin A expression of OSCC cells with the altered Nm23-H1 levels following knockdown and overexpression. By immunohistochemistry, Nm23-H1 expression of metastatic lymph nodes was significantly lower than that of their primary oral cancers, supporting a role of Nm23-H1 in metastasis suppression. Negative Nm23-H1 interpretation of OSCC specimens, in either primary oral cancers or metastatic lymph nodes, indicated a poor survival outcome of patients. On the basis of in vitro studies of Nm23-H1 knockdown and overexpression, we demonstrated an inverse correlation between Nm23-H1 expression and the invasiveness of OSCC cells. Moreover, we observed the concomitant reduction in Nm23-H1 and cyclin A levels of metastatic tumors in both results of in vitro OSCC cells and ex vivo tumor specimens.     

Survivin nm23-H1在非小细胞肺癌中的表达及相关性研究

目的:研究肿瘤相关基因survivin、nm23-H1在非小细胞肺癌﹙NSCLC﹚组织中的表达及相关性.探讨其与临床病理特征之间的关系.方法:采用免疫组化SP 法检测survivin和nm23-H1在52例NSCLC组织、20例对应癌旁组织和15例正常组织中的表达情况.结果:survivin的表达癌组织明显高于癌旁组织的表达,差异具有显著性(P<0.05),正常组织无survivin表达;nm23-H1的表达癌组织明显低于癌旁组织的表达,差异具有显著性(P<0.05);Survivin在NSCLC组织中的表达与年龄、性别、组织类型、淋巴结转移无关(P>0.05), nm23-H1在伴有淋巴结转移的NSCLC组织中表达的阳性率为19.0%(4/21),在未伴有淋巴结转移的NSCLC组织中表达的阳性率为48.4%(15/31),两者差异有显著性;survivin在TNM(Ⅰ+Ⅱ)期NSCLC组织中表达低于在TNM(Ⅲ+Ⅳ)期NSCLC组织中表达;nm23-H1在TNM(Ⅰ+Ⅱ)期NSCLC组织中表达高于在TNM(Ⅲ+Ⅳ)期NSCLC组织中表达,差别有显著意义(P<0.05);survivin在高分化NSCLC中表达低于在低分化NSCLC中表达;nm23-H在高分化NSCLC组织中表达高于在低分化NSCLC组织中表达,差别有显著意义(P<0.05);survivin、nm23-H1在NSCLC中表达负相关(P<0.05).结论:survivin 和nm23-H1 的表达与NSCLC发生、临床进展及预后密切相关.Survivin可以作为NSCLC的早期诊断指标,并可以根据survivin与nm23-H1的联合检测来判断恶性程度及预后.     

Nuclear localization of Nm23-H1 in head and neck squamous cell carcinoma is associated with radiation resistance

BACKGROUND:

Although radiation resistance is a primary issue in radiation therapy, attempts to find predictors of radiation resistance have met with little success. The authors therefore aimed to determine predictors for radiation resistance to improve the prognosis of head and neck squamous cell carcinoma (HNSCC).

METHODS:

HNSCC cell lines, SCC15, SCC25, and QLL1, irradiated with an acute dose of 4 grays (Gy) (RR‐4), a cumulative dose of 60 Gy (RR‐60), and a booster dose of 4 Gy over 60 Gy (RR‐60 + 4), were used with nonirradiated cell lines. Those were used in cDNA microarray, proteomics, Western blotting, and immunofluorescence, respectively. One hundred five HNSCC tissue samples with radiation resistance were analyzed by immunohistochemistry.

RESULTS:

Western blot analysis of RR‐60 cell lines was identical to the data of Nm23‐H1 overexpression by cDNA array and proteomic screening. Immunofluorescence demonstrated significant nuclear translocation of Nm23‐H1 in RR‐4 and RR‐60 cell lines, and less but still intense nuclear shuttling in RR‐60 + 4. Similarly, Nm23‐H1 nuclear localization was observed in 20% (21 of 105) of tissue samples. Univariate analysis demonstrated that Nm23‐H1 nuclear localization was strongly associated with overall and recurrence‐free survival. Multivariate stepwise Cox regression analysis showed that Nm23‐H1 nuclear localization (odds ratio [OR], 7.48) and N stage (OR, 2.13) were associated with overall survival, and Nm23‐H1 nuclear localization (OR, 3.02), T stage (OR, 1.43), and insufficient tumor margin (OR, 3.27) were associated with recurrence‐free survival.

CONCLUSIONS:

Overexpression of Nm23‐H1, specifically its nuclear translocation, may be a powerful predictor of radiation resistance in HNSCC. Cancer 2011. © 2010 American Cancer Society.     

Downstream targets of Nm23-H1: gene expression profiling of CAL 27 cells using DNA microarray

The human nm23-H1 was discovered as a tumor metastasis suppressor based on its reduced expression in melanoma cell lines with low versus high metastatic potential. It encodes for one of two subunits of the nucleoside-diphosphate kinase. Besides its role in the maintenance of the cells NTP pool, nm23 plays a key role in different cellular processes. The role of nm23-H1 in these processes still has to be elucidated. Our goal was to identify Nm23-H1 downstream targets by subjecting Nm23-H1 overexpressing CAL 27 cells oral squamous cell carcinoma (OSSC) to microarray analysis. The genes with changed expression patterns could be clustered into several groups: transforming growth factor beta (TGFbeta) signaling pathway, cell adhesion, invasion and motility, proteasome machinery, cell-cycle, epithelial structural and related molecules and others. Based on the expression patterns observed we presume that nm23-H1 might have a role in OSSCs, which should be confirmed by future experiments.     

NM23-H1 expression of head and neck squamous cell carcinoma in association with the response to cisplatin treatment

We recently reported that low NM23-H1 expression of head and neck squamous cell carcinoma (HNSCC) correlated with poor patients'' prognosis. Growing evidence has indicated that high tumor NM23-H1 expression contributes to a good response to chemotherapy. Therefore, we investigated the role of NM23-H1 in susceptibility of HNSCC cells to cisplatin and its clinical significance, as well as the in vitro study for validation was performed. Using immunohistochemistry, we analyzed NM23-H1 expression in surgical specimens from 46 HNSCC patients with cervical metastases receiving surgery and adjuvant chemoradiotherapy. Low tumor NM23-H1 expression correlated with locoregional recurrence of HNSCC following postoperative cisplatin-basedtherapy (p = 0.056) and poor patient prognosis (p = 0.001). To validate the clinical observation and the effect of NM23-H1 on cisplatin cytotoxicity, we established several stable clones derived from a human HNSCC cell line (SAS) by knockdown and overexpression. Knockdown of NM23-H1 attenuated the chemosensitivity of SAS cells to cisplatin, which was associated with reduced cisplatin-induced S-phase accumulation and downregulation of cyclin E1 and A. Overexpression of NM23-H1 reversed these results, indicating the essential role of NM23-H1 in treatment response to cisplatin. NM23-H1 may participate in HNSCC cell responses to cisplatin and be considered a potential therapeutic target.     

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 nm23-H1 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 immunohisto-chemical staining of nm23-H1 protein in human naso-pharyngeal carcinoma (NPC) was examined, and the relationship between nm23-H1 and both metastasis and prognosis of patients with NPC was also investigated. Methods: Routine LSAB immunohistochemistry with the nm23-H1 monoclonal murine antibody was employed to study the expression of nm23-H1 protein in 95 paraffin-embedded specimens of NPC treated at our hospital. The clinical pathologic data and results of follow-up were also retrieved. Comparisons between patients with and without expression of nm23-H1 protein with respect to metastasis, loco-regional recurrence and survival were performed using Log rank test. Multivariate prognostic analyses were performed by using Cox’s regression model. Results: Nm23-H1 negative expressive tumors were associated with a higher incidence of lymph-node metastasis (86.7%) than those of nm23-H1 positive (48.6%,P<0.01). Nm23-Hl 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 nm23-H1 and prognosis (P<0.01). The expression of nm23-H1 indicated favorable prognosis. Conclusion: It was suggested that nm23-H1 negative expression was significantly associated with lymph-node metastasis, recurrence and distant metastasis. Nm23-H1 may have value for predicting the prognosis of NPC.     

上皮性卵巢癌中nm23-H1基因突变

目的　本文研究nm2 3 H1基因在上皮性卵巢肿瘤中的突变情况 ,以期寻找监测卵巢癌转移并能指导治疗的指标。方法　利用非同位素PCR SSCP技术研究nm2 3 H1基因的 5个外显子的突变情况。结果　正常卵巢、良性卵巢癌及交界性瘤均未发现突变 ,卵巢癌突变率显著增高。低分化程度卵巢癌突变率高于高、中分化程度的卵巢癌。浆液性卵巢癌nm2 3 H1基因突变率高于其他组织学类型。Ⅲ、Ⅳ期卵巢癌突变率高于Ⅰ、Ⅱ期卵巢癌。发生淋巴结转移的卵巢癌突变率高于无淋巴结转移的卵巢癌。结果　nm2 3 H1基因突变容易发生于分化程度较低、有淋巴结转移的晚期卵巢癌病例中。     

p53和nm23-H1蛋白表达与食管癌浸润转移的关系

Objective To study the relationship between expression of p53 and nm23-H1 and differentiation, invasiveness and metastasis in human esophageal carcinoma, and the correlation between expression of p53 and nm23-H1. Methods Expression of p53 and nm23-H1 in 50 patients with squamous cell carcinoma of esophagus was detected by using immuno-histochemical S-P methods. Results 35 cases (70%) and 32 cases (64%) of esophageal squamous cell carcinoma were positive for nm23-H1 protein and p53 protein, respectively. The expression of nm23-H1 was related to lymphatic metastasis (P<0.025), but not related to tumor differentiation, invasiveness, tumor location, tumor length, patient's gender and age (P>0.05). The lymphatic metastasis location positive group had a very lower expression of nm23-H1 and the negative rate was 70.8%, but the negative group had a higher expression and the positive rate was 65.4%. The expression of p53 was related to tumor differentiation and invasiveness (P<0.05), but not related to lymphatic metastasis, tumor location, tumor length, patient's gender and age(P>0.05). Among the three groups, the high differentiation group had the lowest expression of p53 and the positive rate was 29.2%, but the low differentiation group had the highest positive rate (71.4%). As for tumor invasiveness, the group of outer membrane of esophagus infiltrated had the highest p53 protein positive rate (56%), but in the group, of mucous or submucous layer infiltrated p53 protien was not detectable. The low expression of nm23-H1 and the high expression of p53 were also correlated. The expression of nm23-H1 and p53 were both correlated with TNM stage of esophageal carcinoma (P<0.05). The better esophageal carcinomas differentiated, the lower nm23-H1 expressed and higher p53 expressed. Conclusion Low expression of nm23-H1 and high expression of p53 play an important role in the progression of squamous cell carcinoma of esophagus. Nm23-H1 might beta gene markef in the prophecy of patients' prognosis and benefit tumor treatment clinically.