涎腺腺样囊性癌生物学特性研究

由上海第二医科大学附属第九人民医院口腔颌面外科研究完成的“诞腺腺样囊性癌生物学特性研究”课题，于1996年10月通过上海市科学技术成果鉴定。该研究由何荣根、邱蔚六等教授针对涎腺癌的临床和病理实践中提出的和当前国内外争论的热点课题，以腺样囊性癌（ACC）为代表，从其生物学特性认识入手进行了较系统的实验研究。分别建立了来自腭部小涎腺和腮腺的涎腺腺样囊性癌ACC-2和ACC-3细胞系，完成了各种细胞生物学特性检测和鉴定。首次证实ACC细胞在试管内能分泌基底膜成份层粘蛋白（LN）、Ⅳ型胶原、肝硫酸乙酰糖蛋白、内粘蛋白（En…     

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.     

涎腺腺样囊性癌术后放疗47例分析

涎腺腺样囊性癌术后放疗４７例分析孙新东１于亦国２梁超前１孔凡英１满运艳１腺样囊性癌是较常见的涎腺恶性肿瘤，我们于１９８０年１月至１９９０年１２月收治４７例，重点讨论综合治疗的作用及影响预后的因素。１材料与方法全组男２７例，女２０例。年龄２１岁～７３岁...     

唾液腺腺样囊性癌来源的外泌体促进癌细胞的增殖、迁移及侵袭

目的:探索人唾液腺腺样囊性癌不同细胞系分泌的外泌体对其癌细胞增殖、迁移及侵袭能力的影响。方法:使用ExoEasy Maxi Kit (Qiagen,Hilden,Germany)试剂盒提取外泌体,通过透射电镜、纳米粒子跟踪分析（NTA）、蛋白免疫印迹Western blotting技术鉴定提取的肿瘤外泌体。并在外泌体的干预下,采用CCK-8法、平板克隆形成实验、细胞划痕及Transwell实验检测细胞增殖、迁移及侵袭能力。结果:透射电镜及纳米粒子跟踪分析（NTA）检测显示:提取的外泌体主要直径在30~150 nm之间,其形态呈典型的“茶托型”结构;Western blotting检测到外泌体特异性标志蛋白CD9、CD63及TSG101呈阳性表达。CCK-8法及平板克隆实验显示唾液腺腺样囊性癌来源的外泌体可增强其癌细胞增殖能力（P<0.05）。细胞划痕及Transwell实验表明其外泌体可增强癌细胞迁移及侵袭能力（P<0.05）。结论:唾液腺腺样囊性癌不同细胞系均分泌外泌体。唾液腺腺样囊性癌外泌体可增强其癌细胞增殖、迁移及侵袭能力。     

涎腺腺样囊性癌的复发转移因素分析

通过６０例涎腺腺样囊性癌的随诊，结果发现：局部复发率为２１．７％。复发主要与原发部位、病理类型、局部扩散及切除范围有关。腺样囊性癌局部侵袭性强，切除范围不易确定，对术后切缘有癌残留者，应追加术后放疗。本组淋巴结转移少见（３％），转移部位多在原发灶周围。在广泛切除原发灶后，一般无需行预防性颈清扫术。血行转移较多见（２１．７％），常发生于肺部。但出现血行转移的病人经治疗后仍可长期带瘤生存，故血行转移并非切除原发灶的手术禁忌证。     

涎腺腺样囊性癌的治疗体会

作者根据临床经验和文献复习，对涎腺腺样囊性癌的治疗提出以下体会。认为①应当废除那种把腺样囊性癌当作低度恶性处理的老观念，应有足够手术切除；②应当根据不同的原发部位特点，制订不同手术原则；③术后常规放射治疗是需要的；④术前严禁作切开活检，采用术前细针穿活检或术中冰冻切片病理检查；适当病例可采用术后化疗。     

人涎腺腺样囊性癌嗜神经侵袭超微结构的观察研究

目的:通过电镜检测来进一步探索人涎腺腺样囊性癌嗜神经侵袭的超微形态特征.方法:将人涎腺腺样囊性癌Acc-M细胞系注射入5只裸鼠双侧臀部肌肉内,切取其中发生神经侵袭并出现功能障碍的荷瘤;通过透射电镜检测来观察腺样囊性癌的嗜神经侵袭过程. 结果:肿瘤细胞破坏神经外膜及神经束膜,进入神经纤维之间,并以吞噬的方式对神经纤维进行侵袭,致使髓鞘、轴突发生变性、溶解而被吸收.结论:涎腺腺样囊性癌细胞在对神经的侵袭过程中具有一定的吞噬作用,且这种现象在神经侵袭中起了很重要的作用.     

层粘连蛋白与涎腺腺样囊性癌肿瘤生物学特性的关系研究

目的为研究层粘连蛋白与涎腺腺样囊性癌（ＡＣＣ）侵袭和转移的关系。方法用免疫组织化学ＡＢＣ法，观察２５例ＡＣＣ组织中层粘连蛋白（ＬＮ）的表达与分布。结果ＬＮ主要分布在基底膜上以及部分筛状假囊内，腺管内和肿瘤细胞浆内。经统计学检验，发现ＬＮ与病理分型、ＴＮＭ分期和转移有关，即筛状┐管状型、ＴＮＭ早期和无转移者的ＬＮ多阳性表达，而实体型、ＴＮＭ晚期和转移者，ＬＮ多阴性表达。结论ＬＮ表达的高低与ＡＣＣ临床恶性程度侵袭和转移，均有密切关系。     

涎腺腺样囊性癌组织侵袭相关蛋白表达及其与临床因素相关性的研究

目的:观察GFAP、NT-3、Trk及NCAM在腺样囊性癌(ACC)组织中的表达,探讨ACC嗜神经侵袭可能的分子机制,分析4种蛋白表达与临床各因素的关系.方法:采用免疫组化方法检测94例ACC标本中侵袭相关蛋白表达水平,分析4种蛋白在ACC有无嗜神经侵袭组间和临床各因素组间的表达差异.结果:瘤细胞GFAP、NT-3、Trk的过表达和NCAM的低表达在有无嗜神经侵袭和病理分型组间差异均有统计学意义,P均<0.05;NT-3和Trk的过表达在有无复发组间差异均有统计学意义,P均<0.05;NT-3的过表达在有无切缘浸润组间差异有统计学意义,P<0.05.多因素Logistic回归模型分析表明,GFAP、NT-3和Trk的表达与ACC嗜神经侵袭呈正相关,夥直鹞2.097、1.871和3.194,P均<0.05;GFAP表达与ACC病理分型呈负相关,=-1.216,P=0.021.ACC组织中神经组织NCAM的表达高于周围侵袭的肿瘤细胞.结论:瘤细胞过表达GFAP、NT-3和Trk可能共同参与了ACC嗜神经侵袭的过程,低表达NACM对ACC的嗜神经侵袭可能起到了一定的促进作用.     

涎腺腺样囊性癌血道转移的临床分析

目的：探讨腺样囊性癌血道转移问题,期望在临床上对此类病例的治疗引起重视。方法：对６例腺样囊性癌血道转移的病人进行回顾性分析。结果：腺样囊性癌因其生物性特征,术后容易发生局部复发和血道转移。结论：对此类病例强调首期完成根治性手术和进行综合治疗。密切随访,及时发现和治疗转移灶。     

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.

     

Expression of epidermal growth factor receptor in different salivary adenoid cystic carcinoma cell lines

Objective： To investigate the expression of epidermal growth factor receptor, a receptor tyrosine protein kinase, in the subcellular fractions of human salivary adenoid cystic carcinoma cell lines SACC-83 and SACC-LM. Methods： Low metastatic and high metastatic cells of the adenoid cystic carcinoma, SACC-83 and SACC-LM, were cultured. Their subcellular fractions were extracted. The expression of epidermal growth factor receptor was detected with Western blot method, and the results of protein expression were quantitatively analyzed by FluorChem V2.0 software. Results： The results of Western blot analysis indicated that, EGFR expression on the membrane of SACC-83 cells was significantly higher than that of SACC-LM cells, but its expression in cytoplasm was significantly less in the former than the later （P〈0.01）. In SACC-83 cell line, EGFR was over-expressed in membrane （P〈0.01）, but in SACC-LM cell line, EGFR was over-expressed in cytoplasm （P〈0.01）. Conclusion： The results suggest that the obtaining of metastasis ability is related to the high expression of EGFR protein in cytoplasm, so the molecular targeting therapy to EGFR may be an ideal treatment for the invasion and metastasis of salivary adenoid cystic carcinoma.     

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。     

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…     

WIP1 stimulates migration and invasion of salivary adenoid cystic carcinoma by inducing MMP-9 and VEGF-C

The wild-type p53 induced phosphatase 1 (WIP1) is an oncogene overexpressed in a variety of human cancers. Here, we demonstrated that WIP1 silencing reduced MMP-9 and VEGF-C expression as well as migration and invasion of salivary adenoid cystic carcinoma (ACC) cells. Overexpression of MMP-9 or VEGF-C restored migration and invasion in WIP1 knockdown cells, indicating that MMP-9 and VEGF-C are downstream targets of WIP1 signaling. Levels of cyclin D1 and c-Myc, targets of Wnt/β-catenin pathway, were significantly decreased by WIP1 silencing. In addition, WIP1 expression was positively associated with metastasis and prognosis of ACC patients as well as with MMP-9 or VEGF-C in ACC tissues.     

Inhibition of STAT3 reduces proliferation and invasion in salivary gland adenoid cystic carcinoma

In this study, we accessed the expression and correlation of p-STAT3 with Survivin, Cyclin D1, CD147, Slug and Ki67 by immunohistochemical staining of human tissue microarray which contains 72 adenoid cystic carcinoma (AdCC), 12 pleomorphic adenoma (PMA) and 18 normal salivary gland (NSG) using digital pathological scanner and scoring system. We found that the expression of p-STAT3, Survivin, Slug, Cyclin D1 and CD147 was significantly increased in AdCC as compared with PMA and (or) NSG (p<0.05). While, the level of p-STAT3 and expression of Cyclin D1 and CD147 was not associated with pathological type of human AdCC (p>0.05). Correlation analysis of these proteins revealed that p-STAT3 up-regulates the expression of Survivin, Slug, Cyclin D1 and CD147 (p<0.05). Moreover, the activation of STAT3 was associated with proliferation marker Ki-67 (p<0.05). Selective inhibition of STAT3 by a small molecule S3I-201 significantly reduced human SACC-83 and SACC-LM cells proliferation, migration and invasion with the corresponding decrease in expression of Survivin, Slug, Cyclin D1 and CD147. These findings indicate that high phosphorylation level of STAT3 in AdCC is related to Survivin, Slug, Cyclin D1 and CD147. We suggest that the inhibition of STAT3 may be a novel strategy for neoadjuvant chemotherapeutic treatment of AdCC.