摘要
目的胶原蛋白是细胞外基质的主要成分之一,我们在临床数千例恶性胸水转移的非小细胞肺癌细胞胞浆中常见到围绕细胞核或局灶状葱皮排列的纤维样结构,由此我们假设肺腺癌细胞在侵袭转移形成胸腔积液过程中发生与细胞外基质相同的结构变化,即肿瘤细胞胶原化形成"盔甲"式保护膜起到对肿瘤细胞结构及功能的支撑,达到抵御化疗药物、毒素的穿透和放射线的轰击,提高了自我保护能力,适应新的肿瘤细胞增生微环境。方法 TCT方法选取224例肺腺癌细胞学标本,其中胸腔积液标本144例,肺泡灌洗液40例,痰液标本40例,24例为良性胸腔积液作为对照。常规制成细胞包埋块,切片、HE染色及CK7、TTF-1、vimintin的免疫细胞化学染色。透射电镜观察肿瘤细胞内部结构。Masson特殊染色,胶原蛋白Ⅰ、Ⅱ、Ⅲ亚型染色,Western-blot蛋白检测。观察肺腺癌细胞是否存在胶原蛋白亚型及肺腺癌不同标本的表达情况。结果Masson细胞化学染色显示胶原纤维在恶性胸腔积液、支气管肺泡灌洗液及痰肺腺癌标本中的表达率分别为59.7%(86/144)、0%(0/40)、0%(0/40),在良性标本中不表达均为阴性(0/24)。免疫细胞化学结果显示:COL1A1、COL2A1和COL3A1在肺腺癌细胞中的阳性表达率分别为22.6%(38/168),5.4%(9/168),22.6%(38/168),阳性表达主要定位于胞浆,而在增生的上皮细胞中不表达,统计学分析结果显示COL1A1、COL3A1的表达率要显著高于COL2A1的表达率(P=0.024),并且我们发现COL1A1与COL3A1的表达成正相关性(r=0.886,P=0.000)。透射电镜可观察到在肿瘤细胞胞浆中可见多量纤维状物质。结论本研究从形态学上观察到在转移的肺腺癌细胞胞浆内有一些围绕胞核呈葱皮样改变的纤维样结构,结果表明肺腺癌细胞可以产生胶原蛋白,以纤维性胶原Ⅰ,Ⅲ为主,并在转移的肺腺癌细胞中高表达可能与晚期肺癌细胞的抗药性增强有关。
Objective Collagen Protein is one of the main components of extracellular matrix(ECM).We often found arrangement of fibers-onion skin-like structure-surrounding the nucleus or existing in cell cytoplasmpartiy from thousands of clinical cases with malignant pleural effusion of non-small cell lung cancer.So we assumed that adenocarcinoma cells form the same structure as extracellular matrix during invasion and metastasis forming pleural effusion,i.e,the tumor cells generate collagen which is,like ' armor',to support function and structure of tumor cells,to resist chemotherapy drugs,to prevent toxins and radiation,to improve the ability to protect themselves,and to adapt to new microenvironment.Methods TCT screening 224 cases of cytology specimens of adenocarcinoma,including 144 pleural effusion chose,40 alveolar lavage,and 40 sputum specimens;24cases of benign pleural effusion served as controls.Conventional embedding into cell block sections,HE staining,and immunocytochemistry of CK7、TTF-1and vimintin were used.We made observation of the internal structure of tumor cells by TEM.Masson special staining,staining of collagen subtypesⅠ,Ⅱ,Ⅲ,and Western-blot protein detection were made.We observed the expression of collagen subtypes in adenocarcinoma and in different specimens.Results Masson staining showed the expression rate of collagen to be 59.7%(86/144),0%(0/40),0%(0/40)in malignant pleural effusion,bronchoalveolar lavage fluid and specimens of lung adenocarcinoma,and zero in benign specimens(0/24).Immunocytochemistryshowed that the expression rates of COL1A1,COL2A1 and COL3A1in adenocarcinoma cells were 22.6%(38/168);5.4%(9/168),22.6%(38/168),mainly located in the cytoplasm,but not in the proliferated epithelial cells.Statistical analysis showed that the expression of COL1A1,COL3A1 was significantly higher than that of COL2A1(P=0.024),and the expressions of COL1A1 and COL3A1showed positive correlation(r=0.886,P=0.000).Large amounts of fibers were seen in the cytoplasm of tumor cells by TEM.Conclusion This study morphologically observed onion skin-like fiber structures,surrounding the nucleus in adenocarcinoma cell cytoplasm.The results show that lung cancer cells can secrete collagenmainly collagen fibersⅠandⅢ,and their high expression in metastatic adenocarcinoma.may be associated with chemotherapy resistance of advanced lung cancer.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2014年第6期518-524,共7页
Chinese Journal of Histochemistry and Cytochemistry
关键词
非小胞肺细癌
胶原蛋白
上皮间质转化
Non-small cell lung cancer
Collagen
Epithelial-mesenchymal transition
