Reduced expression of p27(Kip1) protein in relation to salivary adenoid cystic carcinoma metastasis.

BACKGROUND: Adenoid cystic carcinoma (ACC) is a malignant tumor of salivary gland origin. It tends to grow slowly but shows frequent recurrence and metastasis, ultimately with a poor outcome. Reduced expression of a cyclin-dependent kinase inhibitor, p27(Kip1), has been reported to correlate with poor survival for patients with various types of carcinoma. However, there has been no previous study reported of p27(Kip1) expression in ACC, to the authors' knowledge. METHODS: To evaluate p27(Kip1) as a prognostic marker, the authors examined the immunohistochemical expression of p27(Kip1) protein in 29 ACCs and correlated its expression with clinicopathologic findings. Eleven pleomorphic adenomas (PAs) were also examined to compare the p27(Kip1) expression in benign and malignant salivary gland tumors. RESULTS: All PAs expressed p27(Kip1) at high levels, whereas 83% of ACCs (24 of 29) showed reduced expression of this protein. Furthermore, the expression levels were significantly lower in ACCs with metastasis than in those without metastasis. The authors also examined the expression of p27(Kip1) in 2 ACC cell lines (ACCh and ACC3) by Northern and Western blot analysis to elucidate the possible mechanism of p27(Kip1) reduction in ACC. Both ACCh and ACC3 expressed p27(Kip1) mRNA, but ACCh did not produce p27(Kip1) protein. In ACCh, the expression of p27(Kip1) protein was induced by treatment with a proteasome inhibitor. CONCLUSIONS: Overall, these findings suggest that reduced expression of p27(Kip1) may correlate with the development and progression of salivary ACC and can be an indicator of its malignant behavior. They also suggest that increased proteasome-mediated degradation may play an important role in this reduction of p27(Kip1) expression.     

Aberrant expression of beta-catenin, Pin1 and cylin D1 in salivary adenoid cystic carcinoma: relation to tumor proliferation and metastasis

The aims of this study were to investigate the expression levels of beta-catenin, Pin1 and cyclin D1 in salivary adenoid cystic carcinomas (SACC ) and to evaluate its clinical importance, furthermore, to elucidate whether beta-catenin expression was aberrant in SACC and whether Pin1 was involved in aberrant beta-catenin and cyclin D1 expression. The expression of Pin1, beta-catenin and cyclin D1 were examined in the specimens of 65 patients with SACC by immunohistochemistry, protein and mRNA expressions were detected by western blotting and RT-PCR in four SACC cell lines. Pin1 was overexpressed in 51 cases of SACC (78%), and high levels of Pin1 expression correlated with cyclin D1 positive expression (p = 0.02). Fourteen (22%) cases showed positive immunoreactivity for beta-catenin protein in the nuclear/cytoplasmic fraction in tumor tissues, which was defined as cytoplasm/nucleus staining, among which quite evident nuclear expression of beta-catenin was detected in six cases (9%), while cyclin D1 positive expression was detected in 41 cases of SACC (63%). Reduced membranous expression of beta-catenin was detected in the cases with metastasis (11/14). Theses results suggest that Pin1 and Wnt signalling pathway are activated in SACC and may play a pivotal role in SACC carcinogenesis and metastasis.     

涎腺腺样囊性癌的复发、转移及治疗初析

目的探讨影响涎腺腺样囊性癌的复发及转移的因素.方法对61例涎腺腺样囊性癌做临床病理分析.结果临床分期晚期位于颌下腺、舌下腺肿瘤神经受侵率高,复发率高;肿瘤复发是死亡的主要原因;局部总复发率为47.5%,远地转移率为29.5%,颈淋巴结转移率为9.8%.结论复发率与发病部位、临床分期及治疗方法等相关,外科是主要的治疗方法,放射治疗可以有效地控制病变的发展.     

NOTCH1 signaling contributes to cell growth,anti-apoptosis and metastasis in salivary adenoid cystic carcinoma

Background: Numerous studies have reported both the tumor-suppressive and oncogenic roles of the Notch pathway, indicating that Notch activity regulates tumor biology in a complex, context-dependent manner. The aim of the present study was to identify the role of NOTCH1 in the cell growth and metastasis of SACC.Methods: We analyzed the expression of NOTCH1 in clinical SACC samples using immunohistochemical staining. We silenced the expression of NOTCH1 and overexpressed activated NOTCH1 to elucidate the effects of NOTCH1 on proliferation, migration and invasion. NOTCH1 target genes were validated by real-time PCR.Results: Our results showed that NOTCH1 was upregulated in SACC tissues when compared with normal tissues, and this upregulation was further enhanced in SACC tissues with metastasis and recurrence when compared with SACC tissues without metastasis. Overexpression of NOTCH1 in SACC cells promoted cell growth, migration and invasion, and knockdown of NOTCH1 inhibited cell proliferation in vitro and tumorigenicity in vivo by inducing cell apoptosis.Conclusions:The results of this study suggest that NOTCH1 plays a key role in the cell growth, anti-apoptosis, and metastasis of SACC. NOTCH1 inhibitors might therefore have potential therapeutic applications in treating SACC patients by inhibiting cancer cell growth and metastasis.     

食管鳞癌nm23-H1蛋白表达及DNA倍体与淋巴结转移关系

目的研究食管鳞癌nm23-H1蛋白表达及DNA倍体状态与区域淋巴结转移的关系.方法应用流式细胞仪检测食管癌患者新鲜手术标本包括食管癌组织(癌)、食管癌旁粘膜(癌旁)、食管正常粘膜(切缘)及食管癌区域淋巴结(淋巴结)中nm23-H1蛋白表达水平及DNA倍体状态.结果无论有、无淋巴结转移,nm23-H1蛋白表达在癌与癌旁、癌与切缘之间差异有显著性(P＜0.05),癌与淋巴结之间差异无显著性(P＞0.05);DNA指数(DI)在癌组织与癌旁、切缘、淋巴结之间比较差异均有显著性(P＜0.05),癌旁、切缘、淋巴结之间DI差异无显著性(P＞0.05);nm23-H1蛋白表达在有、无淋巴结转移两组对应组织间比较差异均有显著性(P＜0.05);两组对应组织比较DI差异无显著性(P＞0.05);DI在不同病理分级间差异有显著性(P＜0.05),DI及nm23-H1蛋白表达在性别、肿瘤部位、病理类型、浸润深度间差异无显著性(P＞0.05).结论nm23-H1蛋白高表达对食管癌转移有一定的抑制作用,结合DNA倍体检测有助于了解食管鳞癌的生物学行为和预后判断.     

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.     

Ezrin在涎腺腺样囊性癌组织中的表达

目的:探讨Ezrin在涎腺腺样囊性癌组织中的表达及意义。方法:用免疫组织化学SP法检测Ezrin在涎腺腺样囊性癌组织和正常涎腺组织中的表达,分析其在涎腺腺样囊性癌组织中的表达与肿瘤的侵袭性、复发、转移和预后的关系。结果:Ezrin在涎腺腺样囊性癌和正常涎腺组织中的阳性率表达分别为44.68%和9.09%,P<0.05;在癌细胞中主要为胞质内表达,染色深,而在正常涎腺组织则主要为细胞膜表达,染色浅或阴性。Ezrin的表达强度与有无神经受侵、有无发生局部复发及远处转移相关,P<0.05。Ezrin表达阳性组和阴性组的5年累积生存率分别为66.70%和100.00%,10年累及生存率分别为27.83%和95.00%,阳性组较阴性组预后差,P<0.05。结论:Ezrin的表达强度与涎腺腺样囊性癌的发生、侵袭性、复发和转移性有关,且Ezrin阳性表达预示患者预后不良。     

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.     

Estrogen receptor expression in salivary gland mucoepidermoid carcinoma and adenoid cystic carcinoma

Estrogen receptor (ER) expression in salivary gland carcinomas is controversial, and most published studies considered no more than 10 cases. We analyzed ER expression by immunohistochemistry in 136 mucoepidermoid carcinomas and 72 adenoid cystic carcinomas. All cases were negative. These results do not support a role for estrogens in salivary gland mucoepidermoid carcinoma and adenoid cystic carcinoma.     

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     

The association between nm23-H1 expression and survival in patients with esophageal squamous cell carcinoma.

The nm23 gene is a potential metastasis suppressor gene originally identified using a murine melanoma cell line. The expression of nm23-H1 protein was examined immunohistochemically in 50 eligible patients with esophageal squamous cell carcinoma (ESCC). The expression was not correlated with other prognostic factors including lymph node metastases; however, overall survival rates of nm23-H1-negative patients were significantly shorter than those of nm23-H1-positive patients (P < 0.05). Furthermore, reduced expression of nm23-H1 was associated with shorter overall survival in patients with involved lymph nodes (P < 0.01), but not in patients without involved lymph nodes. These data support the conclusion that reduced expression of nm23-H1 may be associated with poor prognosis of ESCC patients, suggesting the value of nm23-H1 expression as a prognostic marker for ESCC patients, especially ESCC patients with involved lymph nodes.     

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.     

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).     

涎腺腺样囊性癌来源外泌体促进成纤维细胞PD-L1表达

目的：研究涎腺腺样囊性癌（salivary adenoid cystic carcinoma ,SACC）来源的外泌体（exosome,EXO）对成纤维细胞PD-L1分子表达的影响。方法：通过EXO分离试剂盒提取SACC-83细胞系来源的外泌体并以电镜鉴定其颗粒大小、密度和表型;使用PKH67荧光标记后将EXO与成纤维细胞HPLF共孵育,以共聚焦显微镜观察EXO可否被HPLF细胞摄取;将EXO与HPLF细胞共孵育后,全转录组测序检测筛查该细胞中差异表达基因,GO分析结合KEGG富集分析阐明差异表达基因涉及的生物学功能和相关信号通路;使用qPCR、Western blotting 和流式细胞术检测肿瘤EXO对HPLF细胞中PD-L1、LAG3 和IDO1 在mRNA与蛋白水平表达的影响。结果：SACC-83细胞源EXO特异性表达CD63、CD81和TSG101分子,并可被HPLF细胞内吞摄取;EXO处理HPLF细胞后,细胞中PD-L1 分子显著上调表达（Fold change=10.19）,差异基因显著富集于TNF、NF-κB、cGAS-String 等免疫反应信号通路;体外实验证明EXO可以从mRNA和蛋白水平显著促进HPLF细胞中PD-L1 的表达（24.7±4.75 vs 1.03±0.11,P<0.05）。结论：涎腺腺样囊性癌SACC-83 细胞来源EXO可以促进HPLF细胞中免疫检查点配体PD-L1的表达。     

MTA1、nm23H1 mRNA表达及突变与卵巢癌淋巴结转移的关系

目的 研究MAT1、nm23H1 mRNA表达及突变与卵巢癌淋巴结转移的关系。方法 应用逆转录－PCR（RT－PCR）和逆转录－PCR－单链构像多态性分析（RT－PCR－SSCP）技术,对8例正常卵巢、20例卵巢癌及其相应的20例淋巴结组织,进行MTA1和nm23H1 mRNA表达及突变的检测。结果 转移卵巢癌原发灶MTA1 mRNA高表达率为100％(7/7),无转移为38．5％（5／13）,P＝0．0103;有癌转移淋巴结高表达率为87．5％（6／7）,无癌转移为23％（3／13）,P＝0．0043;有癌转移淋巴结低表达率为100％（7／7）,无癌转移为38．5％（5／13）,P＝0．0102。MTA1／nm23H1 mRNA表达的相对吸光度值（A值,曾称光密度OD值）的比值随转移而增加。单链构像多态性分析（SSCP）未发现突变。结论 MTA1、nm23H1基因的转录表达与卵巢癌淋巴结转移呈正负相关关系,起着下负调控的重要作用。这两个基因的异常表达是卵巢癌转移中的频发事件而与基因突变无关。     

MTA1、nm23H_1 mRNA表达及突变与卵巢癌淋巴结转移的关系

目的　研究MAT1、nm2 3H1mRNA表达及突变与卵巢癌淋巴结转移的关系。　方法　应用逆转录 PCR(RT PCR)和逆转录 PCR 单链构像多态性分析 (RT PCR SSCP)技术 ,对 8例正常卵巢、2 0例卵巢癌及其相应的 2 0例淋巴结组织 ,进行MTA1和nm2 3H1mRNA表达及突变的检测。　结果　转移卵巢癌原发灶MTA1mRNA高表达率为 10 0 % (7/ 7) ,无转移为 38 5 % (5 / 13) ,P =0 0 10 3;有癌转移淋巴结高表达率为 87 5 % (6 / 7) ,无癌转移为 2 3 % (3/ 13) ,P =0 0 118。有转移卵巢癌原发灶nm2 3H1mRNA低表达率为 10 0 % (7/ 7) ,无转移为 30 % (4 / 13) ,P =0 0 0 43;有癌转移淋巴结低表达率为 10 0 %(7/ 7) ,无癌转移为 38 5 % (5 / 13) ,P =0 0 10 2。MTA1/nm2 3H1mRNA表达的相对吸光度值 (A值 ,曾称光密度OD值 )的比值随转移而增加。单链构像多态性分析 (SSCP)未发现突变。　结论　MTA1、nm2 3H1基因的转录表达与卵巢癌淋巴结转移呈正负相关关系 ,起着正负调控的重要作用。这两个基因的异常表达是卵巢癌转移中的频发事件而与基因突变无关。     

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     

nm23-H1基因表达与卵巢癌转移的相关性

背景与目的：转移是卵巢癌治疗失败及患者死亡的首要原因。然而,目前对卵巢癌转移潜能的分子机制知之不多。本研究旨在筛选高频转移卵巢恶性肿瘤细胞,分析卵巢癌高频转移细胞模型中nm23．H1基因表达与肿瘤转移特性的相关性,为系统实验研究和临床实践提供依据。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Northem blot和Westem blot方法测定各类肿瘤细胞nm23 mRNA和蛋白表达水平。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。各类细胞nm23mRNA和蛋白表达水平与肿瘤转移特性呈负相关(r=0．96,P=0．0001)。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。