关键词:上颌窦肿瘤;癌;鳞状细胞;核因子κB
Nuclear FactorKappa B (NFkB ) Expression in Squamous Cell Carcinoma of Maxillary Sinus
LV Hong,WU Lilian,PAN Song
(Department of Otolaryngology,Xianning College First Affiliated Hospital ,Xianning Hubei,437100,China)
ABSTRACT:Objective To investigate nuclear factorkappa B (NFκB) gene expression and its clinical significance in squamous cell carcinoma of maxillary sinus. Methods 42 cases of squamous cell carcinoma of maxillary sinus and 20 mucosa blockes from normal maxillary sinus was assayed using immunohistochemical SP method for detection of NFκB protein expression level. Results NFκB was negative in all 20 normal mucosa specimens, and it was positive in 28 of 42 squamous cell carcinoma of maxillary sinus. The expression of NFκB did not correlate with tumor stage and lymph node metastasis; but correlate significantly with differentiation grade of the carcinoma.A significantly higher positive rate was found in poorly differentiated tumors than in moderate and well differentiated ones. Conclusion NFκB may play an important role in the process of carcinogenesis in mucosa of maxillary sinus and becomes the new target for treating squamous cell carcinoma of maxillary sinus.
KEY WORDS:Maxillary sinus neoplasm; Carcinoma; Squamous cell;Nuclear factorkappa B
核因子κB(NFκB)是1986年由美国麻省理工学院癌症研究中心的Bltimore和麻省Whiteheael生物医学研究所的Rwianse共同发现的[1]。其广泛存在于真核生物中,是一个由复杂的多肽亚单位组成的蛋白家族,是介导细胞内信号传递最重要的核转录因子。NFκB作为信号传导途径中的枢纽,与免疫应答,肿瘤的发生、发展,细胞凋亡的调节以及胚胎发育等有密切联系。
上颌窦鳞状细胞癌是头颈部最常见的恶性肿瘤之一。有文献报道在上皮起源的多种肿瘤如胃癌[2]、膀胱癌[3]等中存在NFκB表达的上调。本研究中我们运用免疫组化技术检测上颌窦鳞状细胞癌和上颌窦正常粘膜组织中NFκB的表达情况,探讨NFκB的表达与上颌窦鳞状细胞癌的发生及临床病理特征的关系。
1 材料和方法
1.1 组织标本
选用我院1989~2004年临床资料完整的上颌窦鳞状细胞癌患者术后保留标本石蜡包块42例,本组患者术前均未行放疗和化疗,所有标本均有完整组织学分级病理资料;再选取上颌窦正常粘膜上皮20例,标本提供者中男12例,女8例,年龄28~70岁。所有标本均用10%甲醛固定,常规石蜡包块,行5μm连续切片。42例上颌窦鳞状细胞癌患者中男25例,女17例,年龄30~72岁,病理诊断为鳞状细胞癌。其中低分化鳞癌13例,中、高度分化鳞癌29例。按1987UICC标准临床分期:Ⅰ期6例,Ⅱ期9例,Ⅲ期21例,Ⅵ期6例。
1.2 免疫组织化学试剂
鼠抗人的NFκB/P65单克隆抗体(SC8008)购自北京中山生物技术有限公司,UltraSensitiveTMSP(鼠、兔)超敏试剂盒(KIT9710)和DAB显色试剂盒(DAB0031)购自福州迈新公司。
1.3 免疫组织化学染色
免疫组织化学染色(SP法)。石蜡切片标本经二甲苯、乙醇脱蜡脱水后,用PBS冲洗。切片浸在已沸腾的枸橼酸盐缓冲液(0.01mol/L pH6.0)中于微波炉持续15min行抗原修复,取出后自然冷却至室温。用PBS冲洗,加3%H2O2在室温下孵育10min,以阻断内源性过氧化物酶。用PBS冲洗,加正常非免疫动物血清在室温下孵育10min。除去血清,加NFκB后于湿盒中在37℃温箱中孵育2h再在4℃冰箱过夜。用PBS冲洗,加生物素标记的二抗在室温下孵育10min。用PBS冲洗,加链霉素抗生素过氧化物酶溶液在室温下孵育10min。用PBS冲洗,加新鲜配置的DAB溶液,显微镜下看见阳