大鼠舌癌变细胞分化异常的表型特征——角蛋白谱分析

目的　阐明口腔癌变细胞分化异常的表型特征。方法　 L SAB免疫组化方法检测 69例 4-硝基喹啉 -1-氧化物 ( 4 NQO)口服诱发大鼠舌癌变各期组织分化标志角蛋白 ( CK) 10 ,13 ,14的表达。结果　随口腔癌变进展 ,CK14自基底层向上逐渐延伸至基底上层表达 ;基底上层 CK13表达逐渐减少 ;基底上层 CK10表达在癌前逐渐上升 ,成癌后则聚然下降。结论　口腔癌变是一种细胞分化异常疾病 ,该过程中角蛋白谱的变化是上皮细胞分化异常的主要表现     

大鼠舌癌变过程中MMP-2及MMP-9基因表达的变化

目的　通过观察基质金属蛋白酶 (MMP)在大鼠舌癌变过程中的不同表达 ,分析MMP在口腔鳞癌 (OSCC)及其癌变过程中的表达规律 ,探讨其与肿瘤发生、发展的关系。方法　用原位杂交方法检测 80例 4NQO饮水诱发大鼠舌癌变各期组织MMP 2和MMP 9基因表达。结果　正常粘膜、轻中度异常增生、原位癌、鳞癌的MMP 2mRNA阳性率分别为 8 3% ,2 2 6 % ,35 7% ,73 9% ;MMP 9mRNA阳性率分别为 8 3% ,19 4 % ,35 7% ,82 6 %。结论　MMP 2、MMP 9基因上调表达与口腔粘膜基底膜破坏密切相关 ,是口腔癌浸润生长的重要分子机制。     

4NAQO诱发大鼠舌癌变细胞分化异常的超微结构观察

目的：探讨4－硝基喹啉－1氧化物（4NQO）诱发大鼠舌癌变细胞分化异常的超微结构及其变化规律。方法4NQO诱发大鼠舌粘膜癌变,透射电镜动态观察癌过程中细胞的超微结构变化。结果：随大鼠舌癌变进势,线粒体则逐渐增多;部分重度异常增生上皮已发生基底膜断裂,原位癌细胞有个别已突破基底膜,结论：口腔癌变是一个渐变过程,鳞状上皮分化异常主要表现为合成产物减少和增代谢活跃细胞器增多,电镜观察基底膜可以早期发现癌变。     

E-钙黏素mRNA在大鼠舌黏膜癌变过程中的表达研究

目的:研究上皮型黏附分子E-钙粘素mRNA在大鼠舌黏膜癌变过程中的表达及其意义。方法:采用4NQO饮水法诱导大鼠舌鳞状细胞癌发生,实时荧光定量PCR技术检测组织标本中E-钙粘素mRNA的表达情况。结果:在大鼠舌鳞状细胞癌发生过程中E-钙粘素mRNA表达降低;上皮单纯增生组、轻度上皮异常增生组、中度和重度上皮异常增生组、鳞状细胞癌组4组标本中E-钙粘素 mRNA的表达量分别是上皮正常组标本中的0.453541倍、0.207062倍、0.190954倍、0.180987倍,且鳞状细胞癌组与上皮正常组间的差异具有统计学意义。结论:在大鼠舌黏膜癌变过程中E-钙粘素mRNA表达随着病理分级的增加呈逐渐降低的趋势。E-钙粘素mRNA表达变化是大鼠舌黏膜癌变过程中的早期事件。     

实时荧光定量RT-PCR检测舌癌E-cadherin mRNA的表达

目的:了解E-cadherin在舌癌中的表达及其临床意义.方法:采用实时定量PCR检测29例舌鳞癌患者的癌组织和正常组织中E-cadherin mRNA,分析E-cadherin基因表达与临床病理参数的相关性.结果:舌癌组织中E-cadherin mRNA表达水平2.23±1.16(E-cadherin/β-actin),低于正常组织组8.59±2.71,两组间差异有统计学意义(P＜0.01),E-cadherin mRNA水平与临床病理参数之间无相关性(P＞0.05).结论:E-cadherin的表达下降是舌癌发生过程中的重要事件,实时定量PCR检测E-cadherinmRNA的表达对舌癌早期诊断有重要参考价值.     

4NQO诱导大鼠舌黏膜癌变过程中HSF1的表达及意义

目的：探讨热休克因子1（ HSF1）在4?硝基喹啉?1?氧化物（4NQO）饮水法诱导大鼠舌黏膜癌变过程中的表达及意义。方法：80只Wistar大鼠随机分两组,实验组用0．001％～0．004％4NQO递增性喂养大鼠8～28周,对照组则用普通自来水喂养,分别于8、16、20、24、28周处死大鼠,通过大体标本、HE染色观察大鼠舌部组织学改变,同时采用免疫组化染色方法观察HSF1在大鼠舌癌变全过程的表达情况。结果：随4NQO作用时间延长和浓度递增,大鼠舌背后部黏膜相继出现颗粒状、白色斑块、疣状突起、菜花状新生物和溃疡状改变。诱导后8、16、20、24、28周,舌癌的发生率分别为0、20．0％、50．0％、66．7％、100％。HSF1在对照组大鼠舌背黏膜上皮中不表达或者微弱表达,而在实验组大鼠随着舌黏膜异常增生程度的增加,HSF1表达明显增强;在原位癌中,HSF1弥漫分布于整个癌巢中;在舌黏膜浸润癌中HSF1在癌上皮中表达有所降低,而在癌基质成纤维细胞中HSF1表达增强。结论：4NQO饮水法可诱导大鼠舌黏膜发生癌变,成功建立大鼠舌癌模型,同时HSF1在大鼠舌癌发生、发展过程中发挥重要作用。     

Osteopontin (OPN) distribution in premalignant and malignant lesions of oral epithelium and expression in cell lines derived from squamous cell carcinoma of the oral cavity

The objectives of this study were to assess the immunolocalization of human osteopontin (OPN) in oral lesions and to identify human cell lines of oral squamous cell carcinoma (OSCC) origin that express OPN mRNA. OPN was localized using immunohistochemistry in the following oral specimens: normal epithelium (n=6), epithelial hyperplasia (n=4), epithelial dysplasia (n=28), carcinoma in situ (n=11) and squamous cell carcinoma (n=43). Cell lines UMSCC-1, MDA TU 138, MDA 686LN, SCC4, SCC9, SCC25, CAL 27 and MDA 1483 were characterized for OPN mRNA expression using Northern blotting. OPN was not detected in normal oral epithelium. Intracellular and intercellular immunore-activity was seen in 75% of hyperplasias, 57% of dysplasias, 54% of carcinoma in situ and 67% of squamous cell carcinomas. UMSCC-1 expressed high levels of OPN mRNA. We conclude that OPN protein is detectable in premalignant and malignant lesions arising from oral epithelium. UMSCC-1 may be a useful cell line in which to conduct in vitro studies designed to clarify the role of OPN in OSCC.     

端粒酶蛋白催化亚基(hTRT)mRNA在口腔鳞癌形成过程中的表达

目的:观察端粒酶催化蛋白基因hTRT在口腔粘膜恶性转化不同阶段中的表达,探讨其与口腔粘膜细胞恶性程度的关系。方法:用原位杂交技术检测82例标本,其中正常口腔粘膜7例、上皮单纯性增生7例、上皮异常增生30例、原位癌8例、口腔粘膜鳞癌30例。结果:正常口腔粘膜、上皮单纯性增生组织中hTRT的mRNA表达较弱,阳性信号仅局限于上皮基底层及副基底层间,阳性率21.4%(3/14);上皮异常增生中hTRTmRNA阳性表达见于多层上皮细胞,并随细胞异形性增高而表达增强,阳性率46.7%(14/30);口腔粘膜鳞癌组织中hTRT的mRNA有较强阳性表达,阳性率81.6%(31/38)。结论:端粒酶hTRT基因的表达与口腔粘膜细胞的恶性程度密切相关,端粒酶的重新激活在口腔鳞癌的形成过程中起了关键性作用     

Homeobox C5 expression is associated with the progression of 4-nitroquinoline 1-oxide-induced rat tongue carcinogenesis

J Oral Pathol Med (2012) 41 : 470–476 Background: Aberrant expression of homeobox genes (HOX), normally required for the differentiation of a particular tissue, has been reported in several types of cancer, but poorly addressed in oral squamous cell carcinoma (OSCC). The present study investigated the expression of HOXC5 in OSCC and identified molecular biomarker whose expression is associated with the multistep oral carcinogenesis. Methods: The expression of HOXC5, proliferation cell nuclear antigen (PCNA), and Bcl‐2 was examined by RT‐PCR and Western blot analysis and confirmed by immunohistochemistry and transferase‐mediated dUTP nick end‐labeling (TUNEL) assay in a 4‐nitroquinoline 1‐oxide (4NQO)‐induced rat tongue carcinogenesis model. Results: Homeobox genes C5 was overexpressed in SCC tissues, but not in normal tissues by RT‐PCR and Western blot analysis. Along with the progress of multistep carcinogenesis, the levels of HOXC5 expression of mRNA and protein significantly increased during the dysplasia (moderate to severe dysplasia) when compared with normal and hyperplasia. The levels of PCNA and Bcl‐2 were sequentially increased from hyperplasia to dysplasia and SCC. By immunohistochemistry, HOXC5 expression was significantly increased in dysplasia, whereas PCNA expression was gradually increased during tongue carcinogenesis. TUNEL‐positive cells were increased until dysplasia, but reduced in SCC. Conclusions: These results indicate that overexpression of HOXC5 is correlated with oral carcinogenesis and strongly contributed to the development of OSCC. HOXC5 may be a useful biomarker and has an emerging therapeutic target of OSCC.     

端粒酶hTR基因在口腔癌前病变及鳞癌中的表达

目的探讨端粒酶ｈＴＲ基因在口腔癌前病变及鳞癌中的表达。方法用原位杂交技术检测８２例标本,其中正常口腔粘膜７例,上皮单纯性增生７例,上皮异常增生３０例,原位癌、鳞癌３８例。结果正常口腔粘膜及上皮单纯性增生组织中ｈＴＲ表达较弱,阳性细胞主要位于基底层,检出率２８．５６％（４／１４）。癌前病变中随着异常增生程度的增加,ｈＴＲ表达逐渐增强,阳性细胞逐步由基底层向棘层波及,检出率６０％（１８／３０）。原位癌及鳞癌均有较强的ｈＴＲ表达,检出率为８１．５８％（３１／３８）。结论端粒酶的激活可能出现在口腔癌前病变的晚期阶段,在口腔鳞癌的形成过程中起了关键性作用。     

4NQO诱发大鼠舌癌变细胞增殖与分化异常的研究

目的 阐明口腔癌变细胞增殖与分化异常的表型特征。方法 ＬＳＡＢ免疫组化方法检测６９例４－硝基喹啉－１－氧化物（４ＮＱＯ）口服诱发大鼠舌癌变各期组织分化标志ｇｐ２３０和增殖标志增殖细胞核抗原（ＰＣＮＡ）的表达。结果 随口腔癌变进展,基底上层ｇｐ２３０表达逐渐减少;基底层ＰＣＮＡ表达逐渐增强。结论 口腔癌变既是细胞增殖异常,又是细胞分化异常疾病。     

iNOSmRNA在正常舌组织、舌白斑、舌鳞癌中的表达及意义

目的　探讨一氧化氮 (NO)在舌鳞癌发生中的作用。方法　原位杂交法检测体内合成NO的关键酶诱导型一氧化氮合酶 (induciblenitricoxidesynthase ,iNOS)mRNA在 1 0例舌正常组织、1 4例舌白斑、68例舌鳞癌中的表达。结果　iNOSmRNA在舌正常组织 ,舌白斑、舌鳞癌中的表达阳性率分别为 2 0 0 %、57 1 %、72 1 % ,各组别间差异均有显著性 (P <0 0 5)。结论　由iNOS诱导产生的NO可能在舌癌的发生中起着重要的作用     

细胞角蛋白19和间隙连接蛋白43在4-亚基硝氧喹啉诱发大鼠舌黏膜癌变过程中表达的研究

目的 研究4-亚基硝氧喹啉（4NQO）诱发大鼠口腔黏膜癌变过程中细胞角蛋白19（CK19）和间隙连接蛋白43（Cx43）的表达,探讨口腔黏膜癌变过程中CK19与Cx43的相关性。方法 利用4NQO诱导SD大鼠的口腔黏膜癌变,运用免疫组织化学的方法检测CK19、Cx43在口腔黏膜癌变过程中各阶段的动态变化。结果 在大鼠正常舌黏膜组织中,CK19阳性染色的细胞散在分布于黏膜基底层;随着大鼠舌黏膜上皮异常增生程度的增加,CK19表达于黏膜基底上层;在口腔鳞状细胞癌（OSCC）组织中,CK19阳性染色细胞分布在黏膜各层。CK19在正常舌黏膜、轻度上皮异常增生、中度上皮异常增生、重度上皮异常增生、OSCC组织中阳性表达率分别为30.00%、50.00%、58.33%、80.00%、91.67%,差异有统计学意义（P<0.05）。在正常舌黏膜中Cx43蛋白主要表达于大鼠舌黏膜上皮细胞的细胞膜上,上皮的基底层、棘层和颗粒层呈阳性染色。随着大鼠舌黏膜上皮异常增生程度的增加,Cx43的表达明显下降。Cx43在正常舌黏膜、轻度上皮异常增生、中度上皮异常增生、重度上皮异常增生、OSCC组织中阳性表达率分别为100.00%、85.71%、66.67%、40.00%、33.33%,差异有统计学意义（P<0.05）。结论 在大鼠舌黏膜癌变过程中,CK19蛋白表达水平随病变程度加重显著升高,提示CK19与口腔上皮细胞的癌变有关;Cx43蛋白表达水平随病变程度加重显著下降,Cx43表达下降是口腔黏膜癌变的早期事件。CK19与Cx43蛋白表达呈负相关,CK19和Cx43的联合检测对OSCC的早期诊断有重要的作用,有利于提高OSCC早期诊断的灵敏度和特异性。     

Reduction of syndecan-1 expression is associated with dysplastic oral epithelium

Syndecans are a family of integral membrane proteoglycans that participate in cell-matrix interactions and growth factor binding. Syndecan-1 expression is induced during keratinocyte differentiation and reduced in squamous cell carcinomas. The purpose of this study was to examine the alteration in syndecan-1 expression in dysplastic oral epithelium. Sixty-six oral biopsy specimens (43 epithelial dysplasias, 3 carcinoma in situ and 20 squamous cell carcinomas) were studied using immunohistochemical methods. The normal epithelium specimens were highly positive for syndecan-1. Fifteen of 46 dysplasias or carcinoma in situ specimens showed negative or weak staining for syndecan-1, two of which were totally negative. Intermediate and strong staining were observed in 17 and 14 dysplasias or carcinoma in situ specimens, respectively. Thirteen (65%) squamous cell carcinomas showed negative or weak staining for syndecan-1, seven of which were totally negative. Only three carcinomas had a strong syndecan-1 expression. Four of the 34 patients with dysplasia who were followed up developed squamous cell carcinoma. All these dysplasias had weak or totally negative syndecan-1 expression. The results suggest that the loss of syndecan-1 is associated with dysplastic changes in oral epithelium.     

整联蛋白连接激酶在口腔白斑及早期浸润癌中的表达

目的 研究整联蛋白连接激酶(integrin-linked kinase,ILK)在口腔自斑及早期浸润癌中的表达特点及演变规律,以期为口腔癌前病变的诊断、治疗及口腔癌的预防提供依据.方法 联合应用免疫组织化学及过碘酸希夫反应(periodic acid Schiff reaction,PAS)方法研究ILK在19例正常口腔黏膜(正常黏膜组)、43例白斑伴上皮单纯增生(单纯增生组)、84例白斑伴上皮异常增生[异常增生组,包括轻、中度异常增生44例(轻中度异常增生组),重度异常增生和原位癌40例(重度异常增牛组)]及54例早期浸润癌(浸润癌组)中的表达情况及分布规律.结果 ILK在正常口腔黏膜中呈阴性表达,在其他3组中的阳性表达率可高达90%(163/181),并且ILK在间质的表达随病变程度的加重而增加(χ2=41.585,P<0.001).白斑从单纯增生至癌变,ILK的表达呈现从上皮浅层向基底层下移的趋势,基底细胞由阴性转为阳性着色,并且伴随从胞质向胞膜的分布改变.重度异常增生组基底层和间质表达之间差异有统计学意义(P=0.029).ILK在早期浸润癌的癌巢表达较癌旁上皮浅,间质阳性的病例[76%(41/54)]较重度异常增生[45%(18/40)]增多.结论 ILK在口腔白斑的癌变中可能起莺要作用,确切机制需进一步研究.     

Immunohistochemical localization of growth factors fibroblast growth factor-1 and fibroblast growth factor-2 and receptors fibroblast growth factor receptor-2 and fibroblast growth factor receptor-3 in normal oral epithelium,epithelial dysplasias,and squamous cell carcinoma

OBJECTIVES: Fibroblast growth factors (FGFs) and their receptors (FGFRs) have been identified in a variety of carcinomas, but there are few studies concerning their presence in oral cancers. The objective of this study was to determine whether FGF-1, FGF-2, and high affinity receptors FGFR2 and FGFR3 are present in the pathogenesis of oral epithelial dysplasias and oral squamous cell carcinoma. STUDY DESIGN: Sections from formalin-fixed, paraffin-embedded samples of oral normal mucosa (n = 14), epithelial dysplasia (n = 20), carcinoma in situ (n = 10), and squamous cell carcinoma (n = 12) were tested for cytoplasmic staining by standard in situ immunohistochemistry with antibodies for FGF-1, FGF-2, FGFR2, and FGFR3. RESULTS: Staining for FGF-1 is decreased or lost in the development of epithelial dysplasia and carcinoma. Staining for FGF-2 showed increased intensity (although not statistically significant) in oral epithelial dysplasias and squamous cell carcinomas and showed a significant increased expression in the upper layers of dysplasias and stratum spinosum-like cells in squamous cell carcinomas. Staining for FGFR2 showed a statistically significant increase in intensity in all layers of epithelial dysplasias and squamous cell carcinomas. Staining for FGFR3 was found in the upper stratum spinosum cells of normal and dysplastic epithelium and well-differentiated squamous cells in squamous cell carcinomas, with a statistically significant increase in staining intensity in dysplastic and carcinomatous tissues. CONCLUSIONS: The loss of FGF-1 is consistent with loss of differentiation in dysplasias and some squamous cell carcinomas. Changes in the localization of FGF-2 and FGFR2 into upper epithelial layers with increasing dysplasia suggest increased mitotic potential of high level cells. The co-localization of FGF-2 and its high affinity receptors in neoplastic tissues suggests an autocrine mechanism of influence on carcinogenesis.     

STAT3 mRNA、EGF mRNA和EGFR mRNA在口腔鳞癌组织中的表达及意义

目的：探讨STAT3 mRNA、EGF mRNA和EGFR mRNA在口腔鳞状细胞癌组织中的表达及其临床意义。方法：采用原位杂交法检测60例口腔鳞癌患者的癌组织、30例不典型增生组织及60例正常口腔黏膜组织中STAT3 mRNA、EGF mRNA和EGFRmRNA的表达,并结合病理分级、有无淋巴结转移等临床指标进行统计学分析。结果：在口腔鳞癌癌变过程中STAT3、EGF和EGFRmRNA的表达在正常口腔黏膜组织、不典型增生组织及癌组织中的表达率均依次增高,分别为25.0%（15/60）、40.0%（12/30）、68.3%（41/60）和33.3%（20/60）、43.3%（13/30）、80.0%（48/60）及18.3%（11/60）、46.7%（14/30）、75.0%（45/60）,组间比较差异有统计学意义（P〈0.05）。在口腔鳞癌组织中STAT3表达与癌的组织学分级、浸润深度及淋巴结转移密切相关（P〈0.05）;EGF表达与癌的组织学分级、浸润深度及淋巴结转移密切相关（P〈0.05）;EGFR表达与癌的组织学分级、浸润深度及淋巴结转移密切相关（P〈0.05）。口腔鳞癌中STAT3、EGF和EGFR mRNA的表达具有相关性。结论：STAT3、EGF和EGFR mRNA可能在口腔癌的浸润、转移及黏膜上皮癌变过程中起重要作用,STAT3、EGF和EGFR mRNA的联合检测可望成为口腔鳞癌早期诊断和判断预后的客观指标之一。     

口腔黏膜癌变过程中丝氨酸/苏氨酸激酶15的表达及意义

目的了解丝氨酸/苏氨酸激酶15（STK15）在口腔黏膜癌变过程中的表达变化,探讨P53/STK15转激活-非依赖通路在口腔鳞癌（OSCC）发生发展中的作用及意义。方法正常口腔黏膜8例,上皮异常增生患者27例,OSCC患者43例, 石蜡包埋组织,采用免疫组化SABC法了解STK15及P53蛋白表达情况,分析二者的相关性及其临床病理学意义。结果STK15在正常口腔黏膜无表达,在上皮异常增生及OSCC中阳性率分别为40.74%（11/27） 和67.44%（29/43）,各组间差异均有统计学意义（P<0.05）;口腔鳞癌中STK15阳性率在P53阳性组高于P53阴性组,在OSCC有淋巴结转移组高于无淋巴结转移组,差异均有统计学意义（P<0.05）。结论STK15过表达是口腔黏膜癌变过程的早期事件,口腔鳞癌STK15过表达可能与p53突变有关并与OSCC淋巴结转移密切相关,P53/STK15转激活-非依赖通路在OSCC发生发展中可能起重要作用。     

小鼠口腔癌淋巴道转移模型骨髓播散肿瘤细胞的检测

目的：检测小鼠口腔癌淋巴道转移模型癌变过程不同时期的骨髓播散肿瘤细胞（disseminated tumor cell,DTC）,探讨小鼠口腔癌变过程与骨髓DTC的关系.方法：病理检查明确诊断为正常舌黏膜、舌单纯性增生的小鼠各10只,舌轻中度异常增生、舌重度异常增生、舌鳞癌的小鼠各20只.采用Ficoll密度梯度离心法提取小鼠股骨骨髓里单个核细胞并制成细胞涂片,免疫细胞化学（Immunocytochemistry,ICC）检测骨髓DTC,比较其数目差异.结果：舌重度异常增生8只、舌鳞癌组13只发现DTC,阳性率分别为40％（8/20）和65 ％（13/20）,每100个单核细胞中DTC数分别为3.03±0.75个和5.20±0.74个,差异均有统计学意义（P＜0.05）.其余各组均未发现DTC（0/10）.结论：小鼠舌黏膜恶变过程中,舌重度异常增生时已出现DTC,并随病变程度加重,骨髓DTC的发生率及其细胞数量增加.