口腔鳞状细胞癌中心体扩增与染色体不稳定的相关性研究

目的 探讨口腔鳞状细胞癌(oral squamous cell carcinomas,OSCC)染色体不稳定形成的潜在机制.方法 采用间接免疫荧光染色法观察8例正常口腔黏膜及32例OSCC石蜡包埋组织的中心体形态及数目,采用流式细胞术检测相应组织DNA倍性情况,分析中心体异常与非整倍体之间的相关性.结果 8例正常口腔黏膜均显示大小数目正常的中心体,32例OSCC组织中,25例有明显的中心体数目扩增及排列紊乱,21例为非整倍体;中心体扩增发生率在非整倍体OSCC组(19/21)高于二倍体OSCC组(6/11)(Fisher确切概率=0.032);相关分析显示中心体扩增与OSCC非整倍体之间存在相关关系(Spearman r=0.413,P=0.019),中心体扩增程度与核型异常程度之间呈直线正相关关系(Pearson r=0.364,P=0.041).结论 中心体扩增作为有丝分裂过程中染色体分离错误的始动因素,可能是OSCC非整倍体形成的潜在原因之一.     

口腔黏膜癌变过程中中心体的扩增及其意义

目的　了解口腔黏膜癌变过程中中心体的状况,探讨中心体的异常在口腔鳞状细胞癌(OSCC)发生发展中的作用及其在口腔黏膜癌变过程中的意义。方法　选取正常口腔黏膜(12例)、轻度上皮异常增生(2例)、中度上皮异常增生(8例)、重度上皮异常增生(12例),口腔高分化鳞癌(10例)、中分化鳞癌(15例)、低分化鳞癌(7例)的石蜡包埋组织,采用间接免疫荧光双重染色(γ-微管蛋白单克隆抗体及细胞角蛋白多克隆抗体)显示上皮细胞中的中心体,分析其在口腔黏膜癌变过程中的变化趋势及在各组间的差异。结果　正常口腔黏膜上皮表现为大小数目正常的中心体,但在72·73%(16/22)的上皮异常增生及84·38%(27/32)OSCC组织中观察到部分上皮或肿瘤细胞中出现异常中心体,表现为中心体直径的增加或数目的增多。具有异常中心体的细胞数目随上皮异常增生程度的增加及肿瘤分化程度的降低而增加,二者存在明显的正相关关系(P<0·01)。结论　中心体的扩增是口腔黏膜癌变过程中的早期事件并与口腔癌发生发展进程有关,口腔黏膜癌变过程中的细胞形态学改变与中心体扩增之间可能存在直接机制上的联系。从调控中心体循环入手治疗口腔癌前病损及OSCC,有可能成为口腔癌预防和治疗的新途径。     

IFNAR1在口腔鳞状细胞癌中的表达及意义

目的：观察Ⅰ型干扰素受体1（type I interferon receptor 1,IFNAR1）在口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）及正常口腔黏膜中的表达,探讨其与患者临床特征及预后的关系。方法：应用免疫组织化学方法检测IFNAR1在108例OSCC及16例正常黏膜组织中的表达水平,分析IFNAR1的表达水平与OSCC临床病理特征的关系。采用SPSS17.0软件包对数据进行统计学分析。结果：IFNAR1在OSCC的表达水平比正常黏膜组织高。IFNAR1的表达水平与T分期、颈淋巴结转移、TNM分期及病理分化程度显著相关（P<0.05）;与患者的性别、年龄、发生部位、吸烟和饮酒无相关性（P>0.05）。IFNAR1表达对OSCC患者生存预后的HR=1.879 （95%CI=0.967-3.651,P=0.063）。Log-rank检验显示,IFNAR1表达高低与OSCC患者预后相关（P=0.057）。结论：IFNAR1在OSCC患者中高表达,其表达上调对OSCC是一个潜在的危险因子,与OSCC患者的预后有相关趋势。     

Kank1在口腔鳞状细胞癌中的表达及意义

目的　研究Kank1在口腔鳞状细胞癌（OSCC）中的表达及意义。方法　使用免疫组化SP法分别检测Kank1在60例OSCC组织及20例正常口腔黏膜（normal oral mucosa,NOM）中的表达情况,并结合OSCC患者的临床相关资料,研究Kank1与OSCC发生发展的关系。结果　Kank1在OSCC组织中的高表达率为55%,而在NOM中的高表达率为100%,两者比较差异有统计学意义（χ2 = 13.585,P＜0.001）;Kank1的表达与OSCC的分化程度有关联,随着分化程度的降低,Kank1的表达也呈下降趋势（P < 0.05）;Kank1在有淋巴结转移的OSCC患者癌组织中的表达明显低于无淋巴结转移的患者（P＜0.05）,但与患者的性别、年龄无明显关联（P＞0.05）。结论　Kank1可能参与了OSCC的发生发展及转移过程。     

p53和增殖细胞核抗原在口腔白斑和口腔鳞状细胞癌中的表达及意义

目的 检测正常口腔黏膜、口腔黏膜白斑和口腔鳞状细胞癌(OSCC)中p53和增殖细胞核抗原(PCNA)表达,探讨p53和PCNA在OSCC癌变过程中的规律及相关性.方法 口腔黏膜白斑标本20例、OSCC标本31例及正常口腔黏膜10例,采用免疫组化SP法检测其p53和PCNA的表达,应用SPSS 11.0软件包对结果进行χ2检验、单因素方差分析和线性回归分析.结果 p53在正常黏膜组、白斑组与鳞状细胞癌组的表达率分别为0%、35%和77.4%,PCNA在上述三组中表达率分别为30%、70%和96.8%,两种表达的组间比较差异有统计学意义(P<0.05);p53蛋白与PCNA表达呈正相关(P<0.01).结论 p53基因和PCNA表达水平均与OSCC发展进程密切相关.     

MDC1蛋白在口腔鳞状细胞癌中的表达及意义

目的：探讨口腔鳞状细胞癌中MDC1（mediator of DNA damage cheekpoint protein1）蛋白的表达水平及意义。方法：采用免疫组织化学Envision法检测10例正常口腔黏膜、60例口腔鳞状细胞癌中MDC1的表达水平．并分析其与临床病理参数之间的关系。结果：MDC1蛋白在口腔鳞状细胞癌中表达高于正常口腔黏膜，两者之间差异有统计学意义（P=0．042），而且MDC1蛋白的表达升高与淋巴结转移有关（P=0．033），但与病理分级（P=0．986）、患者的年龄（P=0．723）、性别（P=0．142）无关。结论：MDC1蛋白在口腔鳞状细胞癌中表达增高，提示它可能在肿瘤的发生和发展中具有一定的作用，为进一步理解细胞DNA损伤与肿瘤发生的关系提供了依据。     

乳腺丝氨酸蛋白酶抑制剂在口腔鳞状细胞癌亚细胞中的表达及意义

目的 检测乳腺丝氨酸蛋白酶抑制剂(mammary serine proteinase inhibitor,Maspin)在口腔鳞状细胞癌亚细胞中的表达,观察其与口腔鳞状细胞癌患者临床病理特征之间的关系,以期为临床提供参考.方法 应用免疫组织化学方法对45例口腔鳞状细胞癌的癌组织标本行Maspin蛋白含量的半定量测定,统计其亚细胞定位表达,并与各临床病理指标进行统计学分析.结果 Maspin蛋白在口腔鳞状细胞癌胞核中表达的强阳性率为24%(11/45),弱阳性率为11%(5/45),阴性率为64%(29/45),细胞核表达与肿瘤大小(P=0.019)、淋巴结转移(P=0.011)及术后转移(P=0.017)呈负相关,与患者术后的生存时间呈正相关(P=0.030);Maspin蛋白在口腔鳞状细胞癌胞质中表达的强阳性率为31%(14/45),弱阳性率为31%(14/45),阴性率为38%(17/45),细胞质表达与淋巴结转移(P=0.038)、术后转移(P=0.004)呈负相关,与患者术后的生存时间呈正相关(P=0.014).结论 Maspin蛋白在口腔鳞状细胞癌胞核或胞质中的表达对判断预后可能具有重要价值.     

口腔鳞状细胞癌p53基因的免疫组织化学研究

随着肿瘤分子生物学研究的不断深入和发展，癌基因和抗癌基因在肿瘤发生和发展中的作用日益受到重视。Ｐ５３抗癌基因的产物是分子量为５３ＫＤ的核磷酸化蛋白质。已证明发生于１３２～２１５位氨基酸残基之间的突变可导致Ｐ５３蛋白失活，并表现出显性的癌基因转化作用［...     

Axl基因在口腔鳞状细胞癌中的表达及意义

目的检测Axl(Anexelekto)在口腔鳞状细胞癌中的表达,探索Axl与口腔鳞状细胞癌发生、发展的关系。方法免疫组织化学法(IHC)检测42例口腔鳞状细胞癌患者的癌组织(口腔癌组)及其相应的癌旁组织(癌旁组)Axl的表达状况。原代培养正常人口腔上皮细胞(HOEC),并培养1株永生化正常口腔角化上皮细胞系(NOK)及2株人舌鳞状细胞癌细胞系(UM1、UM2)。实时荧光定量聚合酶链反应(qRT-PCR)和蛋白免疫印迹分析(Western blot)检测上述4种细胞中的Axl的mRNA和蛋白的表达情况。结果 Axl在口腔鳞状细胞癌组织中的阳性表达率(61.9%)高于其相应的癌旁正常组织(26.2%),差异具有统计学意义(P<0.05),qRT-PCR和Western blot结果示在HOEC、NOK、UM2、UM1中Axl基因的mRNA表达和蛋白表达逐步增高,且在UM2、UM1中明显高于HOEC、NOK。结论 Axl在口腔癌组织及口腔癌细胞株中的表达较口腔正常组织和正常口腔上皮细胞者中高表达,提示Axl基因可能与口腔鳞状细胞癌的发生、发展相关。     

Relationship of p53 overexpression to other cell cycle regulatory proteins in oral squamous cell carcinoma

Aberrations of the p53 gene and the overexpression of its protein are described in a variety of neoplasms, including oral and other head and neck cancers. Here we report the association of p53 (over)expression with a downstream cell cycle inhibitor p21/waf 1 in oral squamous cell carcinoma (SCC). The loss of expression of p16 and p27, two other cyclin-dependent kinase (cdk) inhibitors, was also examined. In this panel of tumours, 10/24 carcinomas were p53-immunopositive. Heterogeneous expression of p21 and p27 was seen in 10/24 SCC and 9/16 SCC, respectively, and this was not correlated to p53 status. The expression of p21 and p27 in these SCCs suggests the existence of mechanisms by which some growing tumour cells may tolerate these cell cycle inhibitors; eight SCCs lacked expression of both inhibitors but only two of these cancers overexpressed p53, suggesting that accumulation of p21/p27 can be independent of the functional status of the p53 gene. Data do not support a clear example of a phenotype that shows an overexpression of p53 with downregulation of p21 or p27 leading to cell cycle alterations. Furthermore, only three SCCs were p16-negative and p53-positive. This suggests that these two tumour suppressors may act in separate pathways.     

Elevated ras p21 expression in oral premalignant lesions and squamous cell carcinomas in Taiwan

Expression of ras p21 oncoproteins was examined in histological sections of oral squamous cell carcinoma (SCC), epithelial dysplasia, epithelial hyperkeratosis and normal oral mucosa using antibodies to ras p21 with an immunoperoxidase technique. Ras p21-positive staining was found in 47 of 51 (92.2%) cases of oral SCC, 4 of 4 (100%) cases of epithelial dysplasia, 7 of 7 (100%) cases of epithelial hyperkeratosis, and 1 of 6 (16.7%) cases of normal oral mucosa. The positive staining rate of ras p21 in oral SCC, epithelial dysplasia or epithelial hyperkeratosis was significantly higher than that in normal oral mucosa (P<0.05). No correlation was found between ras p21 expression and patient age, tumour location, tumour size, clinical staging or histological differentiation of SCC. However, a significant positive correlation was found between ras p21 expression and patients' sex (P<0.05) or regional lymph node status (P<0.05). A significant positive correlation was also discovered between ras p21 expression and patients' smoking habits (P<0.01), as well as daily or total betel quid (BQ) consumption (P<0.05). Of the 47 immunostain-positive SCC patients, specimens from 6 patients were also obtained after chemotherapy, when ras p21 expression was found to be reduced. These results indicate that ras p21 over expression may play an important role in the initiation and progression of oral SCCs in patients who are smokers and BQ chewers.     

Evaluation of immunohistochemical expression of p53, p21, p27, cyclin D1, and Ki67 in oral and oropharyngeal squamous cell carcinoma

J Oral Pathol Med (2012) 41 : 40–46 Background: The purpose of this study was to evaluate whether the immunohistochemical expression of p53, p21, p27, cyclin D1, and Ki67 can predict therapy response and survival in patients with oral and oropharyngeal squamous cell carcinoma treated with preoperative chemoradiation. Methods: Biomarker expression was evaluated by immunohistochemistry in formalin‐fixed, paraffin‐embedded pretreatment biopsies of 111 homogenously treated patients. We assessed the association between clinicopathological variables including response to neoadjuvant chemoradiotherapy as well as the survival of the patients and the expression of the biomarkers as both dichotomized (positive vs. negative) and continuous variables. Results: Biomarker overexpression on the basis of pre‐selected cutoff points was seen in 66 of 111 (59%) cases for p53, in 77 (69%) for p21, in 48 (43%) for p27, in 81 (73%) for cyclin D1, and in 54 (49%) cases for Ki67, respectively. None of the examined biomarkers was able to predict response to neoadjuvant chemoradiotherapy or was associated with survival outcome. Post‐treatment pathologic TNM stage (P < 0.001), pathologic response (P < 0.001), and perineural invasion (P < 0.001) were the only factors having a significant effect on recurrence‐free survival. Post‐treatment pathologic N stage (P = 0.005), post‐treatment pathologic TNM stage (P < 0.001), pathologic response (P < 0.001), and perineural invasion (P = 0.001) had a significant impact on overall survival. Conclusions: Our results suggest that the biomarkers p53, p21, p27, cyclin D1, and Ki67 have no impact on treatment response and survival in patients with oral and oropharyngeal cancer treated with preoperative chemoradiation.     

Cyclin D1 expression in oral squamous cell carcinomas: clinicopathological relevance and correlation with p53 expression

The aim of the present study is to study the relationship between cyclin D1 and the clinicopathological features of oral squamous cell carcinomas. The cyclin D1 and p53 expression in oral squamous cell carcinomas from 56 patients (45 men, 11 women) was studied by immunohistochemistry using monoclonal antibodies. The correlation between cyclin D1 and the clinicopathological features of the oral cancers was evaluated. Cyclin D1 expression was found in 63% of oral squamous cell carcinomas; it was often weak but was more frequently positive in high-grade lesions (P=0.019). The expression was positively correlated with p53 expression (P= 0.06). Radiation therapy did not alter the expression of either cyclin D1 or p53 proteins. Expression of these proteins was not related to the age, gender or survival of the patients, or to stages of the tumors. The cyclin D1 expression was more frequently seen in patients with squamous cell carcinomas of oropharynx, palate, floor of mouth and gingiva. To conclude, cyclin D1 was frequently expressed in oral squamous cell carcinomas. This expression was related to the grade of the tumors and was not similar in various regions in the oral cavity, which may indicate the different tumor biology of cancers from these regions.     

Prognostic role of p21WAF1 expression in areca quid chewing and smoking-associated oral squamous cell carcinoma in Taiwan.

BACKGROUND: Alterations in p21WAF1 protein expression have been observed in a wide variety of human cancers by immunohistochemistry, and both decreased and increased levels of p21WAF1 protein expression have been shown to correlate with poor prognosis. METHOD: To examine the relation between p21WAF1 protein expression and prognosis in oral squamous cell carcinomas (SCCs), we performed an immunohistochemical study with antip21WAF1 antibody on 43 oral SCCs. Immunostaining results were then correlated with p53 protein levels, clinicopathological parameters of the tumors and overall patient survival. RESULTS: Of the 43 patients, 31 (72%) had tumors with positive p21WAF1 nuclear staining and 27 (63%) had tumors with p53 nuclear staining. There was no significant correlation between p21WAF1 and p53 protein expressions and both mutant p53-containing oral SCCs overexpressed p21WAF1 protein. In addition, no significant correlation was found between the p21WAF1 expression and the patients' age, sex, oral habit, cancer location, or primary tumor TNM status at the time of initial presentation. The Kaplan-Meier analysis showed a significant correlation between p21WAF1 protein overexpression and poor patient overall survival (P = 0.049). When p53 and p21WAF1 were evaluated together, the 5-year overall survival was lowest in p53(+)-p21WAF1(+) patients and highest in p53(-)-p21WAF1(-) patients (P = 0.057). CONCLUSION: Combined evaluation of p21WAF1 and p53 expressions may be useful in estimating the prognosis of patients with oral SCCs in Taiwan.     

p53 alterations in betel quid- and tobacco-associated oral squamous cell carcinomas from Taiwan

Alterations of p53 have been explored in Taiwanese oral squamous cell carcinomas (OSCCs) consisting of a betel quid (BQ)/tobacco-related subgroup of 36 subjects and a tobacco-related subgroup of 13 subjects. Mutations in conserved exons were found in 12 tumors. Seven mutations were clustered in a hot-spot region mapped to a region between codons 273–282 in exon 8. The incidence of p53 mutation in BQ/tobacco tumors was 22% (8/36). The frequency of p53 allelic loss (21%, 3/14) in BQ/tobacco tumors approximates to the incidence of mutation. This is the first study demonstrating allelic deletion of p53 in such malignancies. Twenty-four of 43 samples showed positive p53 immunostaining. All tumors harboring mis-sense mutations of p53 in conserved exons exhibited nuclear protein accumulation. The incidence of mutation in conserved exons in BQ/tobacco-associated Asian OSCCs (15%) is significantly different from worldwide OSCCs (46%) related primarily to tobacco consumption ( P =0.00001).     

Fine deletion analysis of 1p36 chromosomal region in oral squamous cell carcinomas

Background:  Squamous cell carcinoma is the most common cancer type of the oral cavity and approximately 50% of the patients succumb to the disease. Unfortunately, few are known about the molecular mechanisms involving in the formation of oral squamous cell carcinoma (OSCC). Recently, it has been reported that 1p36 chromosomal region is deleted in various cancer types and is suspected to harbor various tumor suppressor genes (TSGs). However, limited studies exist on genetics alteration on 1p36 in OSCC and the responsible TSG remained unidentified.
Methods:  To investigate area susceptible to harbor TSG(s) involved in OSCC on 1p36 region, paired normal and tumor tissues of 27 patients with diagnosis of OSCC have been analyzed for loss of heterozygosity (LOH) using nine microsatellite markers based on recent gene mapping.
Results:  LOH was found at least in one locus in 85% of the cases (23 of 27). Interestingly, microsatellite instability was also found in 7% (two of 27) of the cases analyzed. The higher LOH frequencies were found with the markers D1S243 (25%), D1S468 (22%), D1S450 (25%), D1S228 (38%), D1S199 (28%), and D1S1676 (23%).
Conclusions:  Three preferentially deleted regions have been identified in OSCC: region 1 (D1S468-D1S243), region 2 (D1S450-D1S228), and region 3 (D1S199-D1S1676). Multiple candidate TSGs, such as RIZ1, p73, UBE4B, Rap1GAP, EPHB2, and RUNX3, are located in these three areas. The data obtained in this study can be used for further functional analysis of these genes involved in OSCC carcinogenesis.     

p21WAF1/CIP1 expression is a marker of poor prognosis in oral squamous cell carcinoma

BACKGROUND: Previous research on the prognostic relevance of p21(WAF1/CIP1) in oral squamous cell carcinomas (OSCC) yielded inconclusive and contradictory data. OBJECTIVES: To investigate the prognostic significance of p21(WAF1/CIP1) expression, its relationship to p53 accumulation, proliferation-associated proteins Ki-67 and cyclin D1 in relation to survival and clinicopathological features in OSCC. METHODS: Surgical specimens taken from 106 randomly selected patients were studied by immunohistochemistry. Expression of the protein of interest was correlated with clinical data. RESULTS: p21(WAF1/CIP1) expression was found in 61.3% of OSCCs. Expression of p21(WAF1/CIP1) significantly correlated with tumor size (P = 0.005), lymph node involvement (P = 0.002), clinical stage (P < 0.001), and tumor site (P = 0.002). Patients with tumors showing p21(WAF1/CIP1) immunopositivity had decreased 2-year survival (P = 0.018). Expression of p21(WAF1/CIP1) was not related to age, gender, risk factors (tobacco, alcohol), dental status, or tumor differentiation grade. The p21(WAF1/CIP1) expression positively correlated with proliferation-related variables Ki-67 (P = 0.010) and cyclin D1 (P < 0.001), but not with p53 expression. CONCLUSIONS: The expression of p21(WAF1/CIP1) was found to be associated with poorer prognosis and tumor aggressivity in OSCC.     

Association of aberrant p53 and p21WAF1 immunoreactivity with the outcome of oral verrucous leukoplakia in Taiwan

The expression of p53 and p21WAF1 in 53 oral verrucous leukoplakias (OVLs), mostly non-dysplastic lesions, was investigated to ascertain the role of such events in malignant conversion. Immunohistochemical analysis revealed aberrant p53 and p21WAF1 immunoreactivity in 51% (27 cases) and 75% (40 cases), respectively. After an average follow-up period of three and a half years, histopathological examination revealed that 22 (42%) cases had developed oral squamous cell carcinoma (OSCC), 14 (26%) cases had undergone recurrence, and 17 (32%) cases were free of disease. The oncogenic potential of this subset of premalignant lesions warrants attention. A significant difference in the frequency of OSCC progression/recurrence was noted in lesions bearing aberrant immunoreactivity of either p53 (93% vs 42%; P=0.00008) or p21WAF1 (80% vs 32%; P=0.002) in comparison with lesions without immunoreactivity. This study suggested that the aberrant immunoreactivity of p53 and p21WAF1 may represent important alterations of OVL and could affect the outcome of this lesion.