涎腺腺样囊性癌增殖、侵袭及转移相关信号通路的研究进展

涎腺腺样囊性癌是最常见的涎腺恶性肿瘤之一,其生长缓慢、侵袭性强,常沿骨膜和神经扩散,早期即可发生远处转移.涎腺腺样囊性的侵袭和转移是一个多阶段、多因素参与的过程,涉及复杂的机制和多条信号转导通路.文章就相关信号通路的研究进展进行综述.     

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.     

Pim-1 acts as an oncogene in human salivary gland adenoid cystic carcinoma

Background

Pim-1 (Provirus integration site for Moloney murine leukemia virus 1) belongs to the Ser/Thr kinase family and plays a pivotal role in occurrence and development of oncogenesis. Recent studies have demonstrated that Pim-1 phosphorylates RUNX3 and alters its subcellular localization. However, few studies have concerned the implications of Pim-1 in the salivary gland adenoid cystic carcinoma (ACC). In this study, we aimed to clarify the function of Pim-1 in ACC in vitro. Meanwhile, we measured the levels of Pim-1 and RUNX3 in the ACC tissues. The correlations between Pim-1/RUNX3 levels and clinical parameters were also analyzed.

Methods

SACC-83 and SACC-LM cells were transfected with the Pim-1 siRNA. Pim-1 mRNA and protein expression were measured using real-time PCR and immnuoblot, respectively. Cell proliferation was analyzed by CCK-8 assay. Cell cycle, apoptosis, and mitochondrial membrane potential were detected by flow cytometry. Effects of Pim-1 on cells’ invasion were evaluated by transwell migration assay. Pim-1 and RUNX3 levels in ACC tissues were examined by immunohistochemistry.

Results

Pim-1 siRNA reduces cell proliferation, induces apoptosis, causes cell cycle arrest through cell cycle related proteins (Cyclin D1 and CDK4), mitochondrial depolarization, and decreases invasive ability in SACC-83 and SACC-LM cells. Pim-1 and RUNX3 levels are significantly relevant and associated with T-stage and nerve invasion in the ACC tissues.

Conclusions

This study demonstrates the oncogenic role of Pim-1 in ACC. The findings also suggest that Pim-1 may serve as a neoteric therapeutic target and potential prognostic marker for ACC cancer.

     

涎腺腺样囊性癌中丝裂原激活蛋白激酶p38的表达及意义

目的 探讨丝裂原激活蛋白激酶p38(p38MAPK)在涎腺腺样囊性癌(SACC)中的表达及临床意义.方法 采用组织微阵列平台,运用免疫组化和原位杂交技术,检测52例SACC和11例正常涎腺组织中p38MAPK蛋白和mRNA的表达,并分析其与SACC 临床病理特征的关系.结果 正常涎腺组织和SACC组织中,p38MAPK蛋白的阳性表达率分别为36.4%和96.2%,差异有统计学意义(P＜0.01).p38MAPK蛋白表达与SACC的淋巴结转移、神经侵犯有关(均P＜0.05).正常涎腺组织和SACC组织中,p38MAPK mRNA的阳性表达率分别为45.5%和94.2%,差异有统计学意义(P＜0.01).p38MAPK mRNA表达与SACC的淋巴结转移、神经侵犯有关(均P＜0.05).SACC组织中,p38MAPK蛋白与mRNA的表达呈正相关(r=0.409,P＜0.01).结论 在SACC组织中,p38MAPK表达上调,p38MAPK通路可能与SACC的发生、侵袭和转移有关.

Abstract：

Objective To investigate the expression of p38 mitogen-activated protein kinase (p38MAPK) in salivary adenoid cystic carcinoma (SACC) tissues and their clinicopathologic significance.Methods The protein and mRNA expressions of p38MAPK were examined by immunohistochemistry and in situ hybridization, respectively, in 52 cases of SACC and in 11 normal salivary gland tissues adjacent to the tumors on a tissue microarray platform.Results The positive rate of p38MAPK protein expression in the paracancerous normal salivary gland tissues and that in SACC were 36.4% and 96.2%, respectively,showing a significant statistical difference (P ＜ 0.01 ).The protein expression of p38MAPK was positively correlated with the lymph node metastasis and nerve involvement (P ＜ 0.05 ).The positive rates of p38MAPK mRNA in the paracancerous tissues and in the SACC tissues were 45.5% and 94.2%,respectively, with a significant statistical difference (P ＜0.01 ).The mRNA expression of p38MAPK was positively correlated with the lymph node metastasis and nerve involvement (P ＜ 0.05 ).In the SACC, there was a notable positive correlation between the p38MAPK protein and mRNA expressions (r = 0.409, P ＜0.01).Conclusions The expression of p38MAPK is up-regulated in salivary adenoid cystic carcinoma.p38MARK may be involved in the tumorigenesis, development and metastasis of this cancer.     

Cetuximab and platinum-based chemoradio- or chemotherapy of patients with epidermal growth factor receptor expressing adenoid cystic carcinoma: a phase II trial

Background:

Epidermal growth factor receptor (EGFR) is highly expressed in adenoid cystic carcinoma (ACC). The efficacy and toxicity of cetuximab with concomitant platinum-based chemoradio- or chemotherapy in patients with locally advanced or metastatic ACC, respectively, was evaluated.

Methods:

Eligible patients (9 with locally advanced tumour and 12 with metastases) had positive tumour EGFR expression. The cetuximab loading dose (400 mg m⁻²) was followed by 250 mg m⁻² per week. Locally advanced tumours were irradiated (mean dose 65 Gy) and treated with concomitant cisplatin (75 mg m⁻², intravenously). Patients with metastases received concomitant cisplatin and 5-fluorouracil (4 × 1000 mg m⁻²).

Results:

For patients with locally advanced disease (median follow-up: 52 months), the median progression-free survival (PFS) was 64 months and the 2-year overall survival (OS) rate was 100%. For patients with metastases (median follow-up: 72 months), the median PFS and OS were 13 and 24 months, respectively. In both groups the objective response rate was >40%. Skin rash, in-field dermatitis, mucositis and vomiting were the most frequent grade 3/4 adverse events.

Conclusion:

In this single-arm study, the efficacy of cetuximab plus chemoradio- or chemotherapy appeared favourable as compared with historical controls. All side effects were manageable and did not hamper the treatment.     

腺样囊性癌SACC-83细胞对雪旺氏细胞的诱向作用

目的观察并探讨在体外坐骨神经与腺样囊性癌细胞共培养中腺样囊性癌细胞对由坐骨神经长出的雪旺氏细胞生长、迁移的影响。方法采用细胞培养技术将腺样囊性癌细胞、舌癌细胞及成纤维细胞分别与小鼠坐骨神经共培养;制备腺样囊性癌细胞与坐骨神经共培养细胞玻片,并行S100免疫组化染色实验。结果坐骨神经与腺样囊性癌细胞共培养中由坐骨神经长出的雪旺氏细胞向腺样囊性癌细胞群落迁移,表现出趋化性;舌癌细胞与成纤维细胞对雪旺氏细胞无诱向作用;经S100免疫组化染色,坐骨神经生长出长梭形的细胞,染色阳性,为雪旺细胞;多边形的细胞染色阴性,为成纤维细胞。结论坐骨神经与腺样囊性癌细胞共培养中长出的雪旺氏细胞向腺样囊性癌细胞群落趋化性生长,可能是腺样囊性癌嗜神经性重要机理之一,与腺样囊性癌细胞发生雪旺细胞分化有关。     

抑癌基因甲基化与涎腺腺样囊性癌

抑癌基因甲基化是当前肿瘤研究的热点领域,它涉及细胞周期调控、细胞间信号转导、DNA凋亡及肿瘤的侵袭转移等过程.抑癌基因甲基化与涎腺腺样囊性癌的发生及其生物学行为密切相关.     

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。     

Cell type‐dependent biomarker expression in adenoid cystic carcinoma

BACKGROUND:

Adenoid cystic carcinoma (ACC), a rare and progressive salivary malignancy, is characterized by cellular, morphologic, and clinical heterogeneity. The authors of this report hypothesized that the dual cellular composition of ACC plays an important role in biomarker evaluation, tumor biologic behavior, and response to therapy.

METHODS:

To investigate the differential localization and expression of the c‐Kit protein and the epidermal growth factor receptor (EGFR) protein, immunohistochemical analyses were performed on tissue arrays that were constructed from 199 tumors, and the results were correlated with clinicopathologic factors.

RESULTS:

c‐Kit expression was limited to the inner ductal epithelial cells, whereas EGFR expression was limited mainly to the outer myoepithelial cells in the majority of ACCs with tubular and cribriform patterns. In solid ACCs, c‐Kit uniformly was positive, whereas EGFR consistently was negative. A significant statistical correlation was observed between c‐Kit expression and a poor 3‐year outcome, and EGFR expression was correlated with a better 3‐year outcome.

CONCLUSIONS:

The current findings underscored the importance of cellular subtype localization of biomarkers in the clinical and therapeutic stratification of patients with ACC. Cancer 2010. © 2010 American Cancer Society.     

腺样囊性癌组织神经生长因子及受体表达与神经浸润相关性的探讨

目的:观察神经生长因子(NGF)及其功能性受体酪氨酸激酶A(TrkA)在腺样囊性癌(ACC)的表达情况,探讨NGF和TrkA在ACC神经浸润(NI)过程中的作用及其意义。方法:47例石蜡包埋ACC标本,常规HE染色,观察有无NI;并依据形态特点将NI类型分为侵入型和围神经型。免疫组织化学检测NGF和TrkA的表达情况。7例新鲜ACC标本,RT-PCR法检测NGF和TrkA的mRNA的表达水平。结果:47例ACC中37例存在NI,其中侵入型25例、围神经型12例。7例新鲜标本5例存在NI。免疫组织化学染色结果显示,NGF在NI组阳性率为86.49%,非NI组为50.00%,侵入型组为92.00%,围神经组为90.00%;TrkA在NI组阳性率为59.46%,非NI组为40.00%,侵入型组为68.00%,围神经组为50.00%。RT-PCR检测结果显示,NGF mRNA的表达水平在NI组较高,TrkA mRNA的表达两组差异无统计学意义。结论:神经周围癌细胞NGF的表达高于远离神经的癌细胞,而其受体TrkA在神经束膜和肿瘤细胞表面均有表达,NGF与其受体的结合促进癌细胞向着神经聚集并沿其生长,形成NI。     

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.     

Antimetastatic effect of integrin IIb/IIIa inhibitors on salivary adenoid cystic carcinoma

The potential of platelets aggregation by tumor cells was demonstrated by recent studies. Tumor cell metastases were increased by tumor cell-platelet adhesion mediated mainly by integrin IIb/IIIa.[1(6] Salivary adenoid cystic carcinoma (SACC) is one of the commonest malignant salivary gland tumors with a high heamtogenous metastasis rate, and there is a positive correlation between tumor cell-platelet adhesion and metastasist.[7, 8] In this study, adhesion of platelets and SACC cell lines…     

KRAS status and epidermal growth factor receptor expression as determinants for anti-EGFR therapies in salivary gland carcinomas

Salivary gland carcinomas (SGC) are rare cancers with poor prognosis and limited response to conventional chemotherapy. New strategies based on molecular targeted therapy are needed and the EGFR signaling cascade is considered a possible key pathway for therapeutic molecules. We have analyzed 65 SGC of the main histopathological types for the expression of EGFR and and the mutation status of its downstream effector KRAS. EGFR overexpression (+2, +3) has been identified by immunohistochemistry in 75.4%. KRAS mutation analysis was performed by direct genomic sequencing and revealed a KRAS wildtype in 98.5% except of one adenoid cystic carcinoma with a GGT–GAT transition at codon 12 (Gly12Asp). EGFR overexpression and KRAS wildtype are prerequisites for a successful anti-EGFR therapy. The results of this study plead in favor of further therapeutic trials with EGFR-targeting monoclonal antibodies in SGC.     

Mammographic features of adenoid cystic carcinoma

Adenoid cystic carcinoma of the breast accounts for < 1% of breast malignancies. This case report describes the mammographic features and the pathology. Adenoid cystic carcinoma is a well-circumscribed carcinoma that has a relatively good prognosis.     

Analysis and significance of c-MET expression in adenoid cystic carcinoma of the salivary gland

Adenoid cystic carcinoma (ACC), a rare salivary gland malignancy, is a histogenetic, morphologic, and clinical heterogeneous disease. Extensive efforts have been made to characterize molecular events associated with these tumors, including the identification of prognostic and predictive biomarkers. Increased copy number gain and amplification of c-Met, the cell surface receptor for hepatocyte growth factor, has been shown to enhance tumor growth and invasiveness and promote metastasis in certain tumor types. In this study, we evaluated the expression of c-Met by immunohistochemistry (IHC) in a large cohort of salivary gland ACCs and examined its clinicopathologic implications. Archival formalin-fixed paraffin-embedded blocks from 200 ACC patients were used in this study. Pathologic patterns and phenotypic expression of c-Met were recorded and compared with clinical factors including gender, age, disease stage at diagnosis, and clinical outcomes. Correlations between c-MET expression and clinical characteristics were assessed by Pearson''s chi-square test or by the 2-tailed Fisher exact test. Curves describing overall survival were generated by Kaplan-Meier product limit method. Strong c-MET expression was seen in inner ductal and outer myoepithelial cells in 53.2% of the cases. There was no correlation between c-Met overexpression and clinicopathologic parameters or patient''s overall survival ( p = .94074). In conclusion, c-MET expression is high in a significant subgroup of ACC patients. While c-MET expression is not a prognostic factor in ACC, its role as a predictive marker of benefit from MET inhibitors deserves further investigation.     

Primary adenoid cystic carcinoma of the lung: absence of KIT mutations

BACKGROUND: Primary pulmonary adenoid cystic carcinomas (ACCs) are rare lung neoplasms that are challenging to completely resect and can exhibit poor survival. Adjuvant therapy is often ineffective and identification of a targeted novel therapy would be useful. The objective of the current study was to evaluate KIT expression and KIT-activating mutations. METHODS: Primary salivary gland-type tumors of the lung diagnosed between 1972 and 2002 at the Mayo Clinic were identified and the subset of primary pulmonary ACCs were reviewed. Immunohistochemical study for KIT expression and KIT gene mutations in exons 9, 11, 13, and 17 were performed on paraffin-embedded tissue. RESULTS: Forty-nine patients were diagnosed with primary pulmonary ACC. The majority of ACC cases were predominantly the cribriform type (74.4%). KIT immunoreactivity was evaluated in 34 cases and was found to be present in all but 1 case (97%). No mutations were detected in KIT gene exons 9, 11, 13, and 17 in a subset of 12 cases. CONCLUSIONS: Although KIT expression was found frequently in primary pulmonary ACC, a correlation with KIT-activating mutations was not observed.