Inhibitory effects of adhesion oligopeptides on the invasion of squamous carcinoma cells with special reference to implication of αv integrins

We studied invasion-related adhesion events in vitro using three squamous carcinoma cell lines (HSC-3, poorly differentiated type; OSC-19, well-differentiated type; and KB cells, undifferentiated type). An in vitro invasion assay through matrigel in the transwell chamber revealed that HSC-3 cells were most invasive, OSC-19 cells moderately invasive and KB cells least invasive. Inhibition assay of invasion using synthetic peptides RGD, RGDV, RGDS, RGDT, IKVAV and YIGSR, showed that invasion of the three cell lines was significantly inhibited by RGDV. There were other peptides that inhibited invasion significantly including IKVAV for HSC-3, and RGDS and YIGSR for OSC-19. HSC-3 cells and OSC-19 cells adhered to fibronectin, laminin, vitronectin, and type IV collagen, and KB cells did not adhere to laminin but did to fibronectin, vitronectin and collagen type IV. Pretreatment of cells with RGDV peptide in the attachment assay reduced the ability of these cells to bind to vitronectin and fibronectin more efficiently than pretreatment with RGDS. Anti-v antibodies inhibited adhesion of HSC-3, OSC-19 and KB cells to vitronectin, but anti-1 antibodies did not inhibit adhesion. Immunofluorescent microscopic examinations showed that all cell lines were positive for anti-5 and anti-v antibodies, and only HSC-3 cells were positive for anti-3 antibody. 51 was not clearly demonstrated in any of the cell lines. RGDV was the most effective inhibitor of squamous cell carcinoma invasion among the synthetic oligopeptides used in this experiment, and it is suggested that it affects v3-and/or v5-mediated carcinoma cell invasion.Abbreviations BSA bovine serum albumin - MEM Eagle's minimal essential medium - MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide - PBS phosphatebuffered saline - FITC fluorescein isothiocyanate This work supported in part by a grant from the Osaka Cancer Research Foundation     

Analysis of T-cell receptor expressing lymphocytes infiltrating squamous cell carcinomas of the upper aerodigestive tract

T-lymphocytes expressing T -cell receptors (TCRs) of the / type have been suggested to play an important role in mucosal defense against infection and neoplastic transformation. In this study, an immunohistochemical investigation was performed on the distribution of / and / TCRs among tumor-infiltrating lymphocytes. Thirteen patients with squamous cell carcinomas of the upper aerodigestive tract were studied, using monoclonal antibodies and an avidin-biotin-peroxidase technique. Most of the T-cells had an / TCR. Only 1.6% of the T-cells within the cancer tissue and 1.2% of the T-cells in the parenchyma adjacent to the cancer tissue expressed /gd TCRs. These results are consistent with the results of similar studies in bronchial and breast carcinomas. Biopsies from normal oral mucosa in nine healthy individuals showed that 1.3% of the T -cells within the epithelium and 1.0% of those in the lamina propria adjacent to the epithelium expressed / TCRs. Quantitatively the results do not support the theory that / T-cells play an important role in the immunological response against cancer tissue in the mucosa of the upper aerodigestive tract. The functional role of these cells in the mucosa and in response to carcinomas is, however, still uncertain.     

P^16基因蛋白在子宫颈鳞癌组织中的表达

应用免疫组织化学方法对58例子宫颈鳞状上皮细胞癌及5例宫颈上皮内瘤样病变标本的P^16基因表达进行研究,取8例正常子宫颈组织作对照,结果显示：P16基因的阳性表达率在宫颈鳞癌,宫颈上此内瘤样病谱及正常宫颈组织中分别为43．1％,40％,12．5％,肿瘤病理分级越低,临床分期越晚,阳性表达率越低,差异有显著性（P＜0．05）,提示：P16基因的异常表达在子宫颈鳞癌的发生,发展过程中起重要作用。     

Prognostic Significance of Proliferating Cell Nuclear Antigen (PCNA) in Squamous Cell Carcinoma of the Esophagus

Proliferating cell nuclear antigen (PCNA) has been shown tobe of prognostic significance in some gastrointestinal tumors.Immunohistochemical analysis was performed to determine whetherPCNA is useful for predicting the outcome of patients with squamouscell carcinoma of the esophagus. Using a mouse monoclonal antibody,PC 10, the expression of PCNA was studied in resected squamouscell carcinomas of the esophagus from 59 patients who had undergonecurative esophagectomy. None had received any preceding therapy.The proliferation rate was assessed in terms of the percentageof the PCNA-positive nuclear area relative to the total areaof cancer nuclei using a cell analysis system (CAS). Clinicopathologicalvariables including PCNA staining were assessed in relationto prognosis. Survival rate was obtained by the Kaplan-Meiermethod. The PCNA indices (percentage of the positive nucleararea) of the tumors varied from 4.4% to 96.2%. Among the clinicopathologicalvariables, only tumor size (5 cm) and depth of invasion werecorrelated significantly with PCNA index (P<0.05). Microscopically,PCNA was stained in non-keratinized cells but not in keratinizedcells. However the histological grade was not correlated withPCNA index. The survival rate was significantly worse in patientswith high PCNA indices (40%) than in those with low indices(<40%) (P<0.05). However, multivariate analysis revealedthat PCNA index was not an independent prognostic factor.     

喉鳞状细胞癌组织体外原代培养的初步研究

目的 应用体外培养技术,对喉鳞状细胞癌组织进行体外培养,探讨喉鳞癌组织原代培养中的各种影响因素,为建立人喉鳞癌组织的细胞系提供实验基础。方法采用体外组织培养技术,对24例人喉鳞状细胞癌组织进行原代培养,观察原代培养中肿瘤细胞的生长与供体的年龄、肿瘤组织的分化程度及不同培养方法的关系,分析在人喉鳞癌细胞的培养中成纤维细胞、微生物污染的影响。结果 24例人喉鳞状细胞癌组织标本,年龄小于60岁组的细胞生长率为31.25％(5/16例),年龄大于60岁组为37.5％(3/8例);高分化组为100％(2/2例),中分化组为30.8％(4/13例),低分化组25％(2/8例);组织块培养法为43.75％(7/16例),酶消化法培养为10％(1/10例);倒置显微镜下观察,在培养的第5～7天,在贴壁组织块周围可见到有上皮样细胞爬出。全部标本中,成纤维细胞的过度生长和微生物的污染是阻碍人喉鳞癌细胞生长的重要因素。结论培养组织的细胞生长率与供体的年龄关系不大;肿瘤组织的分化程度较高者,细胞的生长率较高;与酶消化分离培养法相比,贴壁组织块培养法的细胞生长率较高;成纤维细胞及微生物的污染是阻碍人喉鳞癌细胞系建立的重要因素。     

舌及口底鳞癌治疗分析

目的 探讨舌及口底鳞癌淋巴转移的手术时机与方式对预后的影响。方法 舌及口底鳞癌67例,非手术组12例(为cN_(2～3)患者),手术组55例。cN_028例行原发灶切除,其中23例行选择性颈淋巴清扫。27例cN_(1～3)(cN15例、cN_(2～3)22例)行原发灶切除加根治性颈淋巴清扫,其中16例cN_(2～3)行岛状肌皮瓣修复术:结果 在34例患者中(cN_(2～3)),手术组5年生存率为54.55％(12/22),非手术组5年生存率为41.67％(5/12)。28例cN_0患者中,4例单纯癌肿切除者术后45～60天出现淋巴转移;23例行选择性颈淋巴结清扫术者,证实13例有隐匿性淋巴结转移,10例未见淋巴结转移。结论 对cN_0患者应尽早行选择性颈淋巴结清扫术。对多区域淋巴结或一个区域多个淋巴结转移的cN_(2～3)患者,手术比非手术组的生存率更高,但后者对提高5年生存质量有一定作用。     

鼻腔鼻窦鳞状细胞癌组织中PCNA和EGFR的表达

目的　探讨增殖细胞核抗原 (PCNA )和表皮生长因子受体 (EGFR )在鼻腔鼻窦鳞状细胞癌组织中的表达 ,并分析其与临床病理特征的关系。方法　采用免疫组化方法 ,对 45例鼻腔鼻窦鳞癌组织进行PCNA和EGFR检测 ,并用 15例正常鼻黏膜作对照。结果　PCNA和EGFR在各级鼻腔鼻窦鳞癌组织中均有表达 ,与正常鼻黏膜比较有显著性差异 ,均有随鳞癌分级的升高而表达增强的趋势。PCNA、EGFR在鼻腔鼻窦鳞癌中的表达有显著的相关性。结论　PCNA和EGFR的表达均能反映鼻腔鼻窦鳞癌的增殖活性 ,表达强度可反映鳞癌细胞增殖活性的高低     

表皮生长因子受体和细胞外信号调节激酶的表达与舌鳞状细胞癌转移关系的观察

目的　了解表皮生长因子受体 (EGFR )和细胞外信号调节激酶 (ERK )在舌鳞状细胞癌中的表达 ,探讨EGFR和ERK与肿瘤增殖、转移的关系。方法　应用免疫组织化学SABC方法 ,分别对 90例舌鳞状细胞癌术后组织标本的EGFR、活化ERK 1/2和Ki 67进行检测 ,并对各因子间的表达关系进行统计分析。结果　在 90例舌鳞状细胞癌组织中 ,2 6例证实有颈部淋巴结转移 ,EGFR、活化ERK 1/2及Ki 67的阳性表达率分别为 93 .3 3 % ( 84/90 )、10 0 .0 0 % ( 90 /90 )和 10 0 .0 0 % ( 90 /90 ) ,阳性细胞分布和反应强度均以肿瘤深部边界区域最为明显 ,统计分析显示EGFR、活化ERK 1/2与肿瘤细胞增殖和肿瘤转移之间呈明显的正相关 (P <0 .0 1)。结论　EGFR、活化ERK 1/2在舌鳞状细胞癌细胞中多呈现过度表达与肿瘤的增殖、转移密切相关     

口腔癌前病变和鳞癌组织中hMSH2蛋白表达的初步探讨

目的:探讨错配修复基因hMSH2的表达与口腔鳞癌发生、发展的关系.方法:采用Western Blot和免疫组化方法检测正常口腔上皮组织、癌旁组织和鳞癌组织中hMSH2的蛋白含量和组织分布.结果:Western Blot检测发现口腔正常上皮组织与癌旁上皮组织的hMSH2表达无明显差异(P＞0.05),而鳞癌上皮内hMSH2蛋白表达较前两者显著增加(P＜0.05).免疫组化结果显示正常上皮内仅见基底层细胞胞核阳性表达;癌旁上皮内hMSH2蛋白表达与异常增生程度有关,异常增生明显的上皮,胞核阳性表达的细胞亦较多;hMSH2蛋白在鳞癌上皮全层细胞胞核内呈阳性表达.所有细胞的胞浆均未见阳性表达.结论:在口腔鳞癌组织中hMSH2蛋白表达水平升高,表明hMSH2的表达在口腔鳞癌的发生、发展过程中可能存在表观遗传调节.     

6例乳腺鳞癌的临床分析

目的：分析乳腺癌的临床特征及治疗方法。方法：回顾性地分析了自1958年建院以来10941例乳腺癌患者中的6例乳腺鳞癌的临床特征，治疗方法及结果。结果：乳腺鳞癌的发病率很低，在我院占所有乳腺癌的0.05％，年龄在42-76岁之间，中位年龄46岁，所有的肿瘤患者均进行手术治疗，绝大部分进行了放疗，只有2例进行了化疗，无瘤生存期差别较大。由于年代较久，有很多项目未做或不清，失访严重。结论；乳腺鳞癌的发病率非常低，确诊需要有较清楚的鉴别诊断，特别是鳞状化生。尚无可推荐的标准治疗方案，有关研究尚应继续进行。