桥粒芯糖蛋白1在不同表皮肿瘤中表达的研究

目的：了解桥粒芯糖蛋白1与表皮肿瘤的病理及生物学行为之间的关系。方法：采用过氧化物酶标记的链霉卵白素免疫组织化学染色方法，检测了桥粒芯糖蛋白1(Dsgl)在鳞状细胞癌、基底细胞癌、Bowen病、日光角化病、角化棘皮瘤、脂溢性角化病及正常皮肤中的表达。结果：Dsgl在正常表皮中显著表达；在鳞状细胞癌和基底细胞癌的肿瘤组织中表达显著减弱或消失；在Bowen病和日光角化病细胞间变区域无表达；在绝大多数角化棘皮瘤、脂溢性角化病中的表达为强而连续的胞膜染色，与正常表皮中的表达相似。结论：皮肤恶性肿瘤中Dsgl的表达显著减弱或消失，可能与肿瘤的侵袭性和转移有关，Dsgl可能对表皮良、恶性肿瘤的鉴别诊断具有一定价值。     

p27、细胞周期素D_1在恶性角质形成细胞肿瘤中的表达

目的探讨恶性皮肤角质形成细胞肿瘤中 p27、细胞周期素 D1的蛋白表达与角质形成细胞肿瘤的关系。方法用免疫组化法检测肿瘤细胞 p27与细胞周期素 D1蛋白表达。结果 p27在 Bowen病和日光性角化病均为高表达,基底细胞癌和鳞状细胞癌为低表达,而且前两者的表达显著高于后两者（ P< 0.05）。细胞周期素 D1在 Bowen病、基底细胞癌及日光性角化病均无表达,而在鳞状细胞癌表达率很高。结论 p27和细胞周期素 D1蛋白表达在皮肤角质形成细胞肿瘤中的改变具有一定肿瘤特异性,可作为判断角质形成细胞肿瘤恶性程度、评估预后的指标。     

银屑病素在光线性角化病、Bowen病及皮肤鳞状细胞癌组织中的表达及意义

目的 研究银屑病素在光线性角化病、Bowen病及皮肤鳞状细胞癌(鳞癌)中的表达.方法 用免疫组化的方法检测18例正常皮肤组织、20例光线性角化病、25例Bowen病、21例高分化皮肤鳞癌及16例低分化鳞癌皮损中银屑病素的表达.结果 银屑病素在正常皮肤组织中表达阳性率为11.1％.在光线性角化病中19例银屑病素表达于角质层至棘层上1～3层,胞质着色,阳性率为95.0％.在Bowen病中22例银屑病素表达于表皮全层角质形成细胞,胞质着色,空泡状细胞包膜及胞质着色,阳性率为88％.在高分化鳞癌中20例银屑病素表达于角质层至棘层全层,角化珠及角化不良细胞着色,阳性率为95.2％;在低分化鳞癌中13例银屑病素表达于角质层至棘层上1～5层,低分化的鳞状细胞不着色,阳性率为92.3％.与正常皮肤组相比,银屑病素在光线性角化病、Bowen病、高分化鳞癌和低分化鳞癌中的表达差异均有统计学意义(P＜0.01).银屑病素在光线性角化病、Bowen病、高分化鳞癌中表达强度逐渐升高,而在低分化鳞癌中表达强度降低,但仍高于正常皮肤组织(P＜0.05).结论 银屑病素在鳞状细胞分化异常的皮肤病中表达异常.     

Smad7在脂溢性角化病、日光性角化病及基底细胞癌中的表达

目的： 检测Smad7在脂溢性角化病、日光性角化病以及基底细胞癌中的表达。方法：对脂溢性角化病、日光性角化病及基底细胞癌标本（各30例）和30例正常标本进行免疫组化染色。结果：23例脂溢性角化病标本、23例日光性角化病标本和28例基底细胞癌标本中Smad7染色阳性,阳性细胞率分别为（31.0±23.0）%,（32.7±26.3）%和（62.6±32.1）%,均显著高于正常组织的（6.7±5.0%）。结论： Samd7可能与脂溢性角化病、日光性角化病以及基底细胞癌的发病有关。     

Ber EP4与EMA染色在皮肤基底细胞上皮瘤和鳞状细胞癌诊断中的意义

目的 观察BerEP4和EMA染色在皮肤基底细胞上皮瘤和鳞状细胞癌诊断中的意义.方法 用免疫组化SP法检测BerEP4和EMA在皮肤基底细胞上皮瘤、鳞状细胞癌、光线性角化病、Bowen病、脂溢性角化病、寻常疣和基底鳞状细胞癌皮损肿瘤成分及周围组织、皮肤附属腺体中的表达.结果 所有基底细胞上皮瘤和基底鳞状细胞癌肿瘤细胞呈BerEP4阳性,而鳞状细胞癌、光线性角化病、Bowen病、脂溢性角化病和寻常疣呈BerEP4阴性;多数鳞状细胞癌、Bowen病和部分光线性角化病肿瘤细胞及病变区域呈EMA阳性,而基底细胞上皮瘤、基底鳞状细胞癌、脂溢性角化病和寻常疣呈EMA阴性.结论 联合使用BerEP4和EMA能很好地协助诊断皮肤基底细胞上皮瘤、基底鳞状细胞癌、癌前病变及一些良性增生性皮肤病.     

Caspase-3在皮肤鳞状细胞癌及光线性角化病中的表达

目的：检测Caspase-3在皮肤鳞状细胞癌及光线性角化病组织中的表达。方法： 应用免疫组化法检测16例皮肤鳞状细胞癌皮损、27例光线性角化病皮损及24例正常皮肤组织中Caspase-3蛋白的表达。结果：Caspase-3在皮肤鳞状细胞癌、光线性角化病及正常皮肤组织的表达率分别为37.50%,51.85%,79.17%,其表达含量在皮肤鳞状细胞癌、光线性角化病、正常皮肤组织逐渐增加。结论：Caspase-3蛋白表达下调可能参与皮肤鳞状细胞癌及光线性角化病的发病过程。     

水通道蛋白3在四种皮肤肿瘤中的表达

目的 探讨水通道蛋白3（AQP3）在四种皮肤肿瘤组织中的表达及意义。方法 应用免疫组织化学方法检测30例脂溢性角化病、15例Bowen病、32例鳞状细胞癌、17例恶性黑素瘤及15例正常人皮肤组织中AQP3的表达。结果 脂溢性角化病、Bowen病、鳞状细胞癌、恶性黑素瘤及正常人表皮组织中均存在AQP3蛋白的表达;脂溢性角化病皮损中AQP3表达水平与正常人对照组差异无统计学意义（P > 0.05）;Bowen病、鳞状细胞癌及恶性黑素瘤皮损中AQP3蛋白表达显著高于正常人对照组（P < 0.01）,其中鳞状细胞癌与恶性黑素瘤的表达最强,均显著高于Bowen病（P < 0.01）,但鳞状细胞癌与恶性黑素瘤比较差异无统计学意义（P > 0.05）。此外,在鳞状细胞癌中AQP3的表达与肿瘤的分化有显著相关性（P < 0.01）;在已转移恶性黑素瘤中AQP3的表达显著高于未转移恶性黑素瘤（P < 0.05）。结论 AQP3在皮肤恶性肿瘤中表达上调。     

Caveolin-1、Met蛋白在日光性角化病、鲍温病和皮肤鳞状细胞癌中的表达

目的：检测Caveolin-1、Met蛋白在肿瘤边缘正常皮肤、日光性角化病、鲍温病和皮肤鳞状细胞癌组织中的表达情况。方法：通过免疫组化SP法，对肿瘤边缘正常皮肤、日光性角化病(Actinic keratosis，AK)、鲍温病(Bowen's Disease，BD)、皮肤鳞状细胞癌(cutaneous squamous cell carcinoma，cSCC)标本中Caveolin-1、Met进行检测。结果：Caveolin-1在正常皮肤、AK、BD、SCC中阳性率依次为16.7%、36.8%、52.9%、78.9%，四组间采用Kruskal-wallis H秩和检验，差异具有统计学意义(P<0.05)。Met在正常皮肤、AK、BD、SCC中阳性率依次为8.3%、26.3%、58.8%、84.2%，四组间采用Kruskal-wallis H秩和检验，P<0.05，差异具有统计学意义。在cSCC组中，Caveolin-1、Met阳性表达之间呈现正相关(r=0.484,P=0.036)。结论：Caveolin-1、Met在AK、BD和cSCC细胞中表达强度与恶性程度相关，且Caveolin-1、Met可能在cSCC发生、发展中具有协同作用。     

5-氨基酮戊酸光动力治疗皮肤肿瘤的疗效观察

目的评价5-氨基酮戊酸光动力(ALA-PDT)治疗日光性角化病、Bowen病、基底细胞癌和鳞状细胞癌的临床疗效。方法将光敏剂5-氨基酮戊酸(ALA)涂于53例日光性角化病、23例Bowen病、36例基底细胞癌、40例鳞状细胞癌患者的皮损处,避光3~4h后,使用光动力治疗仪照射约20min,1次/周,经4~6次治疗后评估疗效。结果 ALA-PDT疗法治疗日光性角化病及Bowen病的有效率为100%;基底细胞癌浅表型的疗程短(3~5次),且有效率为100%,结节型疗程长(4~6次),且有效率仅为77.77%;高分化鳞癌有效率为100%,中分化为85.71%,低分化鳞癌有效率为40.00%,所有皮肤肿瘤患者的总痊愈率为84.21%,有效率为95.35%。结论 ALA-PDT治疗日光性角化病、Bowen病、浅表型基底及高分化鳞状细胞癌疗效确切,美容效果佳,无不良反应,治疗结节性基底细胞癌及中分化、低分化鳞状细胞癌尤其是年老体弱、美容要求高的患者亦有一定的疗效。     

抗增殖蛋白2在皮肤鳞状细胞癌中的表达及其对A431细胞增殖的影响

目的：观察抗增殖蛋白2在皮肤鳞状细胞癌、鲍温病、日光性角化病组织中的表达情况及其对A431细胞增殖作用的影响。方法：采用免疫组织化学方法检测抗增殖蛋白2在40例皮肤鳞状细胞癌、18例鲍温病、17例日光性角化病组织以及9例正常皮肤组织中的表达水平。采用CRISPR-Cas9法构建两组靶向敲除抗增殖蛋白2的皮肤鳞状细胞癌A431细胞模型（KO299组及KO320组）,使用Western印迹法证实其敲除效果,通过CCK8法及细胞克隆形成实验分析其对细胞增殖的影响。使用Western印迹法对AKT蛋白表达及其ser473位点磷酸化产物水平进行分析。 结果：抗增殖蛋白2在皮肤鳞状细胞癌(90.00％)、鲍温病(66.70％)、日光性角化病(41.18％)的表达高于正常组织(0.00％)（均P<0.05）,皮肤鳞状细胞癌抗增殖蛋白2表达高于日光性角化病(P<0.05)。在不同性别、年龄、部位、病程、肿瘤直径、浸润深度、分化、Broder分级的皮肤鳞状细胞癌组织中抗增殖蛋白2的表达差异无统计学意义（均P>0.05）。两组抗增殖蛋白2敲除组A431细胞的蛋白表达水平、细胞活力、克隆形成数显著低于对照组 (均P<0.05)。p-AKT表达随抗增殖蛋白2表达水平的降低而显著下调（均P<0.01）。结论：抗增殖蛋白2的表达可能促进了皮肤鳞状细胞癌的发生,并且其机制可能与促癌基因AKT蛋白活化相关。     

SGK1在日光性角化病、基底细胞癌及鳞状细胞癌中的表达

目的：检测SGK1在日光性角化病（AK）、基底细胞癌（BCC）及鳞状细胞癌（SCC）中的表达。方法：采用免疫组化SABC法检测SGK1在25例正常皮肤(NS)、25例AK、28例BCC、28例皮肤鳞状细胞癌标本中的表达。结果：NS、AK、BCC和SCC标本中,SGK1阳性细胞率分别为（40.03±14.42）%,（36.63±14.28）%,（52.82±18.73）%和（52.58±20.13）%。BCC组和SCC组分别与NS组比较,差异均有统计学意义（Ps<0.05）。各组SGK1染色阳性细胞率>50%的标本分别为6例（24%）,3例（12%）,16例（57.14%）和14例（50%）,BCC组和SCC组分别与NS组比较,差异均有统计学意义（Ps<0.05）。结论：SGK1的高表达可能与基底细胞癌及鳞状细胞癌的发病有关。     

蛋白激酶D1及其磷酸化位点在皮肤鳞状细胞癌、Bowen病及光线性角化病组织中的表达

目的 探讨蛋白激酶DI(PKD1)及其磷酸化位点pPKD1-tyr463和pPKD1-ser916在鳞状细胞癌(SCC)、Bowen病和光线性角化病(AK)中的表达及意义.方法 收集新鲜SCC、Bowen病、AK及正常皮肤组织各10份,RT-PCR法检测各组样本中PKD1在基因水平的表达,Western印迹法检测各组样本中PKD1及其磷酸化位点在蛋白水平的表达.另收集蜡块组织SCC 50份、Bowen病20份、AK 20份及正常表皮组织10份,免疫组化检测PKD1、pPKD1-tyr463及pPKD1-ser916的表达情况.结果 正常皮肤组织、SCC、Bowen病和AK组织中PRKD1 mRNA的表达量分别为0.64±0.09、5.37±1.06、2.69±0.72和2.43±0.46,4组间差异有统计学意义(F=21.37,P＜0.05),且SCC、Bowen病和AK组织的表达水平均显著高于正常组织(P＜0.05),SCC组织又显著高于AK和Bowen病组织(均P＜ 0.05),而Bowen病与AK组织的表达量差异无统计学意义(P＞0.05).PKD1总蛋白及pPKD1-tyr463在SCC和Bowen病组织中主要表达在棘层细胞及异形细胞的细胞质和细胞膜,且阳性表达率均显著高于正常皮肤组和AK组(均P＜0.01);pPKD1-ser916仅在部分高分化SCC癌巢中少量表达,而低分化鳞癌、AK、Bowen病及正常皮肤组织中均未见表达;SCC组中PKD1阳性表达率随鳞癌病理分级的提高而增加,且PKD1与pPKD1-tyr463的表达呈正相关(rcc=0.479,P＜0.05).Western印迹检测结果与免疫组化检测结果大致相符.结论 PKD1及其磷酸化位点Tyr463可能参与复层鳞状上皮来源的皮肤肿瘤的形成和进一步发展分化,在皮肤SCC形成进程中PKD1可能通过Tyr463位点活化而发挥促进作用.     

NUAK2 localization in normal skin and its expression in a variety of skin tumors with YAP

BackgroundNUAK2 is a critical gene that participates in the carcinogenesis of various types of cancers including melanomas. However, the expression patterns of NUAK2 in normal skin and in various types of skin tumors have not been fully elucidated to date.ObjectivesTo elucidate the distribution and localization of NUAK2 expression in normal skin, and characterize the expression patterns of NUAK2 and YAP in various types of skin tumors.MethodsIn this study, we characterized the expression of NUAK2 in tissues by developing a novel NUAK2-specific monoclonal antibody and using that to determine NUAK2 expression patterns in normal skin and in 155 cases of various types of skin tumors, including extramammary Paget’s disease (EMPD), squamous cell carcinoma (SCC), Bowen’s disease (BD), actinic keratosis (AK), basal cell carcinoma (BCC) and angiosarcoma (AS). Further, we analyzed the expression patterns of YAP and p-Akt in those tumors.ResultsOur analyses revealed that NUAK2 is expressed at high frequencies in EMPD, SCC, BD, AK, BCC and AS. The expression of p-Akt was positively correlated with tumor size in EMPD (P = 0.001). Importantly, the expression of NUAK2 was significantly correlated with YAP in SCC (P = 0.012) and in BD (P = 0.009).ConclusionsOur results suggest that the YAP-NUAK2 axis has critical importance in the tumorigenesis of SCC and BD, and that therapeutic modalities targeting the YAP-NUAK2 axis may be an effective approach against skin tumors including SCC and BD.     

磷酸化Stat3和磷酸化Akt及磷酸化ERK1/2在表皮良恶性肿瘤中的表达

目的 探讨磷酸化细胞信号传导与转录活化因子3(p-Stat3)和磷酸化Akt(p-Akt)及磷酸化细胞外信号调节激酶(p-ERK1/2)在表皮良恶性肿瘤中的表达.方法 采用免疫组化的方法检测脂溢性角化病,光线性角化病,鲍恩样丘疹病,基底细胞上皮瘤,鲍恩病,鳞状细胞癌中p-Stat3,p-Akt及p-ERK1/2的表达.结果 p-Stat3,p-Akt及p-ERK1/2阳性表达率在表皮恶性肿瘤中明显高于良性肿瘤(P均<0.05).p-Stat3和p-Akt的阳性表达率在表皮恶性肿瘤中从高到低依次为鳞状细胞癌>鲍恩病>基底细胞上皮瘤,而p-ERK1/2的阳性表达率无差异(P>0.05).在表皮恶性肿瘤中,p-Stat3和p-Akt的阳性表达呈显著正相关(P<0.05).而p-ERK1/2的阳性表达与p-Star3、p-Akt的阳性表达之间无相关性(P>0.05).结论 p-Stat3,p-AKT和p-ERK1/2参与表皮恶性肿瘤的发病.     

Immunohistochemical study of cyclooxygenase-2 and p53 expression in skin tumors

Overexpression of cyclooxygenase-2 (COX-2) has been demonstrated in various cancers, including experimentally promoted tumors, gastrointestinal cancers, breast tumors and skin tumors. The mechanism that controls COX-2 expression is not yet clear. Currently, it is reported that COX-2 expression is frequently associated with mutated p53 genes. The goal of this study was to evaluate the expression patterns of COX-2 and p53 in several skin tumors and their correlation. An immunohistochemical method was used to investigate the expression of COX-2 and p53 proteins on formalin-fixed, paraffin-embedded tissue specimens of squamous cell carcinomas (SCC), basal cell carcinomas (BCC), Bowen's disease (BD), actinic keratosis (AK) and porokeratosis. The expression of COX-2 increased in 50% (5/10) of SCC, 80% (8/10) of BCC, 40% (4/10) of BD, 50% (5/10) of AK, and 20% (2/10) of porokeratosis cases. The expression of p53 increased in 90% (9/10) of SCC, 70% (7/10) of BCC, 70% (7/10) of BD, 50% (5/10) of AK, and 40% (4/10) of porokeratosis cases. COX-2 positivity rates of the p53-positive skin tumors were 56%, 100%, 57%, 80% and 25% in SCC, BCC, BD, AK and porokeratosis, respectively. However, the correlation between p53 and COX-2 expression in skin tumors was not statistically significant ( P  > 0.05). Our results indicate that skin COX-2 and p53 may play roles in skin tumors, but that there is no apparent correlation between the two markers.     

紧密连接蛋白Claudin-1、7在日光性角化病和皮肤鳞状细胞癌中的表达

目的：检测紧密连接蛋白Claudin-1、7在日光性角化病(AK)和皮肤鳞状细胞癌（SCC）中的表达水平。方法：收集2014年11月至2016年11月我院病理科AK和SCC组织标本各30例,选取癌旁正常皮肤组织30例作为对照组,采用免疫组化方法检测癌旁正常表皮、AK和SCC组织中Claudin-1、7蛋白的表达。结果：对照组、AK和SCC组织中Claudin-1蛋白细胞阳性率分别为100%,53.33%和13.33%,三组两两比较,差异均有统计学意义(Ps<0.05)。Claudin-7在癌旁正常表皮、AK和SCC组织中均呈阴性表达。结论：Claudin-1蛋白在癌旁正常表皮、AK和SCC组织中表达水平逐渐下降,Claudin-1蛋白表达下调可能与表皮肿瘤的恶性转化有关。     

Increased expression of an epidermal stem cell marker, cytokeratin 19, in cutaneous squamous cell carcinoma

Background Cytokeratin 19 (CK19) has been considered to be a putative marker for epidermal stem cells in the hair follicle bulge. Cumulative reports have shown that epidermal stem cells play an important role in skin carcinogenesis. However, to date there has been no report on the clinical alteration of the stem cells in squamous cell carcinoma (SCC). Objectives To investigate alteration of the stem cells and proliferating cells and to assess their relationship and potential contribution to SCC. Methods Thirty paraffin‐embedded neoplastic skin lesions, consisting of 10 cases each of actinic keratosis (AK), Bowen disease (BD) and SCC, were examined immunohistologically for CK19 and Ki‐67. Results Positive reactivity for CK19 was seen in 30% of AK, 50% of BD and 80% of SCC lesions. There was significantly higher expression levels of CK19 in SCC than in AK and BD (P < 0·05). In addition, BD lesions harboured a significantly higher number of CK19‐positive cells than did AK lesions (P < 0·05). There were significant differences in Ki‐67 labelling indices between AK and BD and between AK and SCC (P < 0·001), but not between BD and SCC (P > 0·05). Furthermore, a serial section comparison study showed that there was a minor population of cells co‐expressing CK19 and Ki‐67 in a subset of the tumour cells of SCC samples. The percentage of CK19+ cells significantly correlated with that of Ki67+ cells in all examined neoplastic skin lesions. Conclusions These results suggest that CK19 expression may be associated with the retention of stem cell characteristics or a state that is uncommitted to terminal squamous differentiation.     

Cyclooxygenase-2 expression and angiogenesis in squamous cell carcinoma of the skin and its precursors: a paired immunohistochemical study of 35 cases

BACKGROUND: Cyclooxygenase (COX)-2 expression and tumour-induced angiogenesis appear to be increased in squamous cell carcinoma (SCC) of the skin. In other cancers, COX-2 is a pro-angiogenic factor. The association between angiogenesis and COX-2 has not been studied in skin cancer. OBJECTIVES: To assess the onset of increased COX-2 expression and angiogenesis in the multistage carcinogenesis of SCC as well as the correlation between those two parameters. PATIENTS/METHODS: We performed a retrospective paired immunohistochemical analysis of normal skin, actinic keratosis (AK), Bowen's disease (BD) and SCC among 35 individuals. Specimens were considered COX-2 immunopositive when 5% or more of the tumour cells showed clear evidence of immunostaining. To quantify active angiogenesis, we used a Ki-67-CD34 double-labelling immunohistochemical stain and calculated the fraction of proliferating endothelial cells. The Chalkley method was used to determine the microvessel density. To detect hypoxia, a carboanhydrase IX immunostain was used. RESULTS: Compared with normal epidermis (0%), AK (31%), BD (22%) and SCC (40%) were significantly more likely to be COX-2 immunopositive (P < 0.01). The fraction of proliferating endothelial cells and the Chalkley scores paralleled multistage carcinogenesis (P < 0.05 between different stages). COX-2 immunopositivity was fairly correlated with hypoxia and higher proliferating endothelial cell fractions but not with Chalkley counts. CONCLUSIONS: Induction of COX-2 expression and angiogenesis are both early events in the development of SCC. In addition to ultraviolet light, hypoxia and COX-2 may be involved in skin tumour angiogenesis.     

An assessment of the malignant potential of actinic keratoses and Bowen's disease: p53 and PCNA expression pattern correlate with the number of desmosomes

Actinic keratoses (AK) and Bowen's disease (BD), both intraepidermal skin tumors, have a potential progression to squamous cell carcinoma (SCC). To evaluate the malignant potential of AK and BD, the expression pattern of p53 protein and proliferating cell nuclear antigen (PCNA) were examined in five types of AK and BD by immunohistochemistry. The ultrastructural difference of epidermal cells between AK and BD lesions was investigated. In the study of p53 and PCNA expression, the atrophic and acantholytic types of AK showed lower positive rates compared to others. These two types did not demonstrate all layers expression pattern. The number of desmosomes of the epidermal cells was significantly reduced in BD, and in the bowenoid and hypertrophic types of AK compared with other types of AK The number of hemi-desmosomes showed greatest reduction in BD and the bowenoid type of AK On the basis of our findings, it is hypothesized that atrophic and acantholytic types of AK may have the lowest, and the bowenoid type of AK and BD may have the highest, malignant potential.