皮肤鳞状细胞癌中基质金属蛋白酶2及金属蛋白酶组织抑制因子2的表达

目的：探讨皮肤鳞状细胞癌(简称鳞癌)中基质金属蛋白酶2(MMP-2)、金属蛋白酶组织抑制因子2(TIMP-2)的表达及其与肿瘤分化程度和淋巴结转移的关系。方法：用ABC免疫组化技术观察50例皮肤鳞癌(高分化鳞癌34例，低分化鳞癌16例，其中14例淋巴结转移)。结果：MMP-2与TIMP-2均表达于瘤细胞和癌旁间质细胞、血管内皮细胞质中。MMP-2表达阳性率为76％(38／50)，在低分化鳞癌组中的表达明显高于高分化组(P=0．021)和有淋巴结转移组(P=0．003)。TIMP-2表达阳性率为74％(37／50)，在高分化或无淋巴结转移的鳞癌组中的表达明显高于低分化(P=0．010)或有淋巴结转移组(P=0．018)。结论：皮肤鳞癌中MMP-2和TIMP-2表达与其分化程度及淋巴结转移有关。     

角质形成细胞生长因子及其受体反义寡核苷酸对HaCaT细胞的影响

目的研究角质形成细胞生长因子(KGF)及其受体(KGFR)反义寡核苷酸对KGF介导的细胞周期和凋亡变化的抑制作用。方法利用永生化角质形成细胞株-HaCaT细胞进行体外培养,流式细胞仪检测KGF引起的HaCaT细胞细胞周期和凋亡变化以及KGFR mRNA反义寡核苷酸对KGF的抑制作用。结果 10ng/mL KGF刺激24 h后可使S期比率较自然生长组增加15%(P＜0．05),并伴凋亡增加2．9%(P＜0．01)。针对KGFR mRNA不同位点的2个序列反义寡核苷酸作用后,S期比率和凋亡率降低,且抑制作用呈浓度依赖性(P均＜0．05)。反义寡核苷酸作用后G_0G_1期相应增加,也呈浓度依赖性(P＜0．01)。各实验组G_2M期比率差异无统计学意义(P均＞0．05)。结论 KGF可能是造成银屑病KC增殖和凋亡的诱因。KGFR mRNA反义寡核苷酸对KGF介导的HaCaT细胞增殖、凋亡增加具有抑制作用。     

凋亡抑制蛋白livin在人皮肤鳞状细胞癌中表达的初步研究

目的:研究凋亡抑制蛋白livin在人皮肤鳞状细胞癌(简称鳞癌)组织及细胞中的表达情况.方法:①采用免疫组化染色方法检测18例鳞癌组织中livin的表达情况;②利用反转录(RT)-PCR及Western blot的方法检测皮肤鳞癌细胞系A431细胞中livin mRNA及蛋白的表达情况.结果:①livin蛋白在正常皮肤组织中不表达,鳞癌组织中livin蛋白的阳性表达率为72.2%;②livin基因mRNA及蛋白在皮肤鳞癌细胞系A431中表达较高.结论:凋亡抑制基因livin在人皮肤鳞癌组织及细胞中高表达,提示livin基因在皮肤鳞癌发生发展中起一定作用,为皮肤鳞癌的诊断及治疗研究提供实验基础.     

皮肤鳞状细胞癌组织中垂体瘤转化基因和碱性成纤维细胞生长因子蛋白及mRNA的检测与分析

目的 探讨垂体瘤转化基因(PTTG)和碱性成纤维细胞生长因子(bFGF)在皮肤鳞状细胞癌(皮肤鳞癌)中的作用及其临床病理意义.方法 采用原位杂交及免疫组化的方法检测42例皮肤鳞癌组织及42例正常皮肤组织中PTTG、bFGF蛋白及mRNA的表达.结果 皮肤鳞癌组织中PTTG和bFGF蛋白(27/42、64.3％和31/42、73.8％)及mRNA(25/42、59.5％和29/42、75.0％)阳性表达率均显著高于正常皮肤组织蛋白(5/42、11.9％和9/42、21.4％)及mRNA表达(3/42、7.1％和7/42、16.7％),组间比较差异均有统计学意义(P＜0.05):PTTG蛋白和mRNA与皮肤鳞癌的组织学分级密切相关(P＜ 0.05);PTTG与bFGF的蛋白及mRNA表达均呈正相关(P＜0.05).结论 PTTG和bFGF蛋白及mRNA表达率升高可能与皮肤鳞癌的发病相关.     

基质金属蛋白酶2及血管内皮生长因子皮肤鳞状细胞癌中的表达及其临床意义

目的 探讨皮肤鳞状细胞癌(简称鳞癌)中基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)、血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达及其与肿瘤分化和淋巴结转移的关系.方法 用ABC免疫组化技术检测50例皮肤鳞癌标本(其中高分化鳞癌34例,低分化鳞癌16例,伴有淋巴结转移14例)中MMP-2、VEGF表达.结果 MMP-2、VEGF主要表达在癌巢中心,表达阳性率分别为76%、72%;二者在低分化鳞癌中的表达明显高于高分化者(P<0.05),且其表达越高,淋巴结转移率越高(P<0.05).结论 皮肤鳞癌中MMP-2和VEGF表达与其分化程度及淋巴结转移有关.     

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.     

p27和细胞周期蛋白E在Bowen病及皮肤鳞状细胞癌中的表达

目的 探讨p27和细胞周期蛋白E在Bowen病及皮肤鳞状细胞癌（鳞癌）发病中所起的作用。方法 用免疫组化S-P法检测p27和细胞周期蛋白E在16例Bowen病、35例皮肤鳞癌和25例正常人皮肤中的表达。结果 p27在Bowen病和皮肤鳞癌中的表达均低于正常人皮肤，在Bowen病、高分化鳞癌和低分化鳞癌中的表达随肿瘤分化程度的降低呈下降趋势。细胞周期蛋白E在Bowen病和皮肤鳞癌中的表达均高于正常人皮肤，在Bowen病、高分化鳞癌和低分化鳞癌中的表达随肿瘤分化程度的降低而增强。在Bowen病和皮肤鳞癌中，p27和细胞周期蛋白E的表达呈负相关。结论 p27异常低表达和细胞周期蛋白E异常高表达可能与Bowen病和皮肤鳞癌的发病有关。     

p53、Gadd45α蛋白在皮肤鳞状细胞癌、基底细胞癌中的表达

目的:检测p53和Gadd45α蛋白在皮肤鳞状细胞癌和基底细胞癌组织中的表达.方法:应用免疫组化法对30例皮肤鳞状细胞癌和25例基底细胞癌组织中的p53和Gadd45α蛋白表达进行检测.结果:p53蛋白在皮肤鳞状细胞癌和基底细胞癌组织中的表达阳性率分别为56.67%、48%,与正常皮肤组织比较差异均有显著性(P=0.006;0.025);Gadd45α在皮肤鳞状细胞癌和基底细胞癌中表达分别为43.33%、52%,均高于正常皮肤组织的表达(P=0.031;0.010).高分化皮肤鳞状细胞癌组阳性表达率为75%,高于中低分化组的22.22%(P=0.008).结论:皮肤鳞状细胞癌和基底细胞癌组织中p53、Gadd45α异常表达可能参与了发病过程.     

c—jun c—fos在尖锐湿疣和宫颈鳞状细胞癌发病中的作用

目的 探讨c-jun、c-fos在宫颈鳞状细胞癌、尖锐湿疣发病中的作用。方法 采用免疫组化方法和RT－PCR方法检测c-jun、c-fos mRNA和蛋白在宫颈鳞状细胞癌及宫颈尖锐湿疣、外阴尖锐湿疣的表达情况，PCR方法检测标本中HPV感染情况。结果 宫颈鳞状细胞癌、宫颈尖锐湿疣、外阴尖锐湿疣中c-jun、c-fos mRNA表达增高。宫颈鳞细胞癌、宫颈尖锐湿疣及外阴尖锐湿疣中c-jun蛋白的表达分别为47．8％、21．0％、64．7％；c-fos蛋白的表达分别为63．0％、89．5％、88．2％。结论 HPV感染促进了c-jun、c-fosmRAN和蛋白在宫颈尖锐湿疣和宫颈鳞状细胞癌中表达，低危型HPV中其蛋白表达高于高危型HPV感染。     

皮肤鳞状细胞癌皮损中hTERT mRNA的表达

目的探讨hTERT基因表达在皮肤鳞状细胞癌(鳞癌)进展过程中的作用。方法收集10例正常皮肤、11例假上皮瘤样增生(PEH)、66例皮肤鳞癌原发灶及12例淋巴结转移灶标本和临床、病理学资料,采用原位杂交法检测hTERT mRNA,结果以图像分析方法定量分析。结果hTERT mRNA在正常皮肤、PEH及鳞癌中的阳性系数(PU)分别为0.84±0.58、3.06±1.09和10.01±0.60(P<0.01),鳞癌与前两者间差异有非常显著性(P<0.01);高分化与低分化组PU值分别为8.52±0.63和13.42±1.02,差异有非常显著性(P<0.01);转移组与无转移组、已发生转移病例原发灶与转移灶间差异均无显著性(P>0.05)。结论与正常皮肤、PEH相比,鳞癌中hTERT基因表达明显增高,分化越低表达越强,提示端粒酶激活在鳞癌恶变过程中起了关键作用。     

Hyaluronan,CD44 and versican in epidermal keratinocyte tumours

BACKGROUND: The high molecular weight polysaccharide hyaluronan is a major component of the extracellular matrix between the vital cells of human skin epidermis. The levels of hyaluronan, and those of the hyaluronan receptor CD44 and the hyaluronan binding proteoglycan versican, correlate with the aggressiveness of different human carcinomas of epithelial origin. OBJECTIVES: To study skin keratinocyte tumours for the expression of hyaluronan, the hyaluronan receptor CD44 and the hyaluronan binding proteoglycan versican. METHODS: Paraffin-embedded sections of 114 basal cell carcinomas (BCC), 31 in situ carcinomas (ISC) and 35 squamous cell carcinomas (SCC) were stained with a hyaluronan specific probe, biotinylated hyaluronan binding complex, and with monoclonal antibodies against CD44 and versican. RESULTS: Compared with normal epidermis, ISC and well differentiated SCCs showed an enhanced hyaluronan signal on carcinoma cells while CD44 expression level resembled that of normal skin. Less differentiated SCCs showed reduced and irregular expression of both hyaluronan and CD44 on carcinoma cells. In BCCs, hyaluronan and CD44 signals were absent or very low on the surface of carcinoma cells. However, hyaluronan was frequently present on BCC cell nuclei, a feature completely absent in ISC, SCC and normal epidermis. An accumulation of hyaluronan in the connective tissue stroma around the tumour was more frequent in SCCs than BCCs. Versican staining was positive around hair follicles and dermal blood vessels of normal skin. Peritumoral versican signal was present in a part of the BCCs but not in other tumours. CONCLUSIONS: The completely different hyaluronan and CD44 expression patterns in BCC and SCC probably reflect the different origins of the tumours, with BCC an undifferentiated keratinocyte and SCC a keratinocyte at an early stage in the differentiation pathway. The difference in hyaluronan and CD44 expression between these tumours may also contribute to the difference in their capacity to metastasize.     

皮肤鳞状细胞癌和基底细胞上皮瘤膜型基质金属蛋白酶-1、基质金属蛋白酶-2表达

目的 探讨皮肤鳞状细胞癌(简称鳞癌)和基底细胞上皮瘤(简称BCE)膜型基质金属蛋白酶-1(MT1-MMP)、基质金属蛋白酶-2(MMP-2)表达及其与临床病理特征的关系。方法 应用原位杂交和ABC免疫组化技术检测48例皮肤鳞癌和41例BCE手术切除标本中MT1-MMP、MMP-2表达。结果 鳞癌中MT1-MMP、MMP-2表达明显高于BCE(P<0.05),且二者的表达呈显著正相关(r=0.370,P<0.05)。MT1-MMP表达在高分化或伴有淋巴结转移的鳞癌中明显上调(P<0.05),而MMP-2表达增加仅见于伴有淋巴结转移的鳞癌(P<0.05)。鳞癌和BCE中MT1-MMP、MMP-2表达与患者性别、年龄、病程、发病部位、原发/转移病灶及BCE侵袭性无关(P>0.05)。结论 MT1-MMP、MMP-2表达上调与皮肤鳞癌的淋巴结转移有关。MT1-MMP、MMP-2表达水平不能区别BCE是否具有侵袭性。     

热休克蛋白10、60在皮肤鳞状细胞癌、基底细胞癌、日光性角化病中的表达

目的 探讨热休克蛋白（HSP）10、60在皮肤鳞状细胞癌（SCC）、基底细胞癌（BCC）和日光性角化病（AK）中的表达水平。方法 采用免疫组化EnVision两步法测定HSP10、60在皮肤SCC、BCC、AK中的阳性表达水平,并与正常组对照。结果 与对照组比较,HSP10组只有BCC组的阳性表达高于正常组（Z = 3.24,P < 0.01）,AK组（Z = 0.74,P > 0.05）和SCC组（Z = 0.52,P > 0.05）与对照组比较差异无统计学意义;HSP10组中AK与BCC,AK与SCC的差异有统计学意义（P < 0.05）,但SCC与BCC组间差异无统计学意义（P > 0.05）。HSP60组三组的阳性表达均高于正常组,其中AK（Z = -2.90,P < 0.01）、BCC（Z = -2.15,P < 0.05）、SCC（Z = -2.78,P < 0.01）;三组间两两比较结果为AK = SCC > BCC（P < 0.05）。结论 HSP60的高表达可能与鳞状细胞癌、日光性角化病的生物行为有关。     

Expression of EGF receptor,involucrin, and cytokeratins in basal cell carcinomas and squamous cell,carcinomas of the skin

Summary The distribution of several markers of keratinocyte differentiation was studied in normal epidermis, basal cell carcinomas (BCCs), and squamous cell carcinomas (SCCs) using the immunoperoxidase technique on frozen sections of punch biopsy specimens. As markers a panel of chain-specific monoclonal antibodies (MoAbs) directed against cytokeratin (CK) 4, 8, 10, 13, 18, and 19, a polyclonal antiserum against involucrin, as well as a MoAb against the epidermal growth factor (EGF) receptor were used. In 15 out of 19 BCCs tested, expression of CK 8 was seen. Only a few individual cells in a limited number of BCCs showed positive staining for CK 4, 18, or 19. No expression of CK 10 was seen except for some foci of cell keratinization. Involucrin was not found in BCCs except for some squamous horn cysts. In all BCC cells expression of EGF receptor was found. In the suprabasal layers of normal epidermis from SCC patients, positive staining for CK 10 was seen. A few individual cells in a limited number of SCCs showed positive staining for CK 4, 8, or 18. Involucrin was expressed in the center of SCCs and in the upper layers of normal epidermis. Expression of EGF receptor was found in all SCC cells. These results demonstrate differences in cellular origin and differentiation between BCC and SCC.     

Immunophenotypic analysis of the p53 gene in non-melanoma skin cancer and correlation with apoptosis and cell proliferation

BACKGROUND: Sunlight precipitates a series of genetic events that lead to the development of skin cancers such as basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). The p53 tumour suppressor gene, which plays a pivotal role in cell division and apoptosis, is frequently found mutated in sunlight-induced skin tumours. OBJECTIVE: To investigate the immunoreactivity of the p53 gene in non-melanoma skin cancers and to correlate its expression with apoptotic and cell proliferation markers. METHODS: We analysed 35 non-melanoma tumours including 19 BCCs and 16 SCCs from sun-exposed skin areas. p53 protein expression was studied immunohistochemically using the DO7 monoclonal antibody against wild-type and mutant p53 forms. The percentage of p53-immunopositive nuclei was measured by image analysis. Cell proliferation and apoptosis were also assessed by image analysis following Ki-67 immunostaining and application of the TUNEL method on paraffin sections, respectively. RESULTS: The percentage of p53-expressing cells varied from 3.5 to 90 in BCCs (median value 54.4%) and from 3.7 to 94 in SCCs (median value 40.3%). The mean value of Ki-67-positive cells was comparable in both groups of tumours with a mean value of 40.6% in BCCs and 34.6% in SCCs. Conversely, the TUNEL assay showed sporadic staining of apoptotic cells within the tumours with a mean value of 1.12% in BCCs and 1.8% in SCCs. p53 protein expression was correlated positively with cell proliferation (r = 0.75, P = 0.000001) and negatively with apoptosis (r = -0.23, P = 0.05). CONCLUSION: p53 immunoreactivity was high in the majority of the skin carcinomas examined and correlated positively with cell proliferation and negatively with apoptosis. The p53 protein overexpression appears to be related to an inactivated protein resulting from mutations of the p53 gene or other unclear molecular mechanisms.     

Lamin expression in normal human skin,actinic keratosis,squamous cell carcinoma and basal cell carcinoma

BACKGROUND: Aberrant expression patterns of nuclear lamins have been described in various types of cancer depending on the subtype of cancer, its aggressiveness, proliferative capacity and degree of differentiation. In general, the expression of A-type lamins (lamins A and C) has been correlated with a non-proliferating, differentiated state of cells and tissues. OBJECTIVES: To establish and compare the expression patterns of lamins in normal human skin, actinic keratosis (AK), squamous cell carcinoma (SCC) and basal cell carcinoma (BCC). METHODS: Expression patterns of the individual lamin subtypes were studied immunohistochemically. The proliferation capacity of the tumour cells was detected using a specific antibody to Ki-67, and was related to the A-type lamin expression patterns. RESULTS: In normal skin, lamin A was expressed in the suprabasal cell compartment of the epidermis, whereas the basal cells were mostly unstained. BCCs and SCCs stained positive in most cells, while the epidermis overlying BCC and SCC and the epidermis in AK stained homogeneously and strongly in the basal cells in addition to the suprabasal cells. Lamin C was expressed in some basal cells of normal epidermis while the suprabasal cells stained strongly positive. Both BCCs and SCCs stained strongly positive for lamin C, with the difference that in BCC the staining was predominantly present in nucleolar structures with occasional staining of the nuclear envelope. The epidermis overlying SCC showed strong positivity in the lamina of virtually all cells. The expression of lamin C in the basal cells of AK resembled the expression pattern seen in the epidermis overlying BCC, i.e. a nucleolar staining next to nuclear envelope staining. Lamin B1 and B2 were found in virtually all cells in normal epidermis, AK, BCC, SCC and the epidermis overlying cancer. The percentage of Ki-67-expressing cells was highest in BCC (45%), and gradually decreased via epidermis overlying BCC, AK, SCC, and epidermis overlying SCC, to normal skin (11%). Simultaneous expression of A-type lamins and Ki-67 occurred in approximately 50% of the proliferating (Ki-67 positive) cells in BCC and SCC. CONCLUSIONS: Significant changes occur in the expression patterns of A-type lamins in both premalignant and malignant lesions of the skin. The profound overlap of lamin A and Ki-67 staining patterns indicates that the proliferating tumour cells may obtain a certain degree of differentiation. Finally, lamin A expression in the basal cell layer of the apparently normal epidermis overlying BCC may suggest its involvement in the primary process.     

iNOS在皮肤肿瘤中的表达及其意义

目的:检测诱导型一氧化氮合酶(iNOS)在皮肤良恶性肿瘤中的表达。方法:采用免疫组织化学技术检测25例脂溢性角化病、25例光线性角化病、25例基底细胞癌、30例鳞状细胞癌(I级13例,II-III级17例)、10例正常皮肤组织中iNOS的表达。结果:2例(8.00%)脂溢性角化病、13例(52.00%)光线性角化病、11例(44.00%)基底细胞癌、22例(73.33%)鳞状细胞癌中iNOS呈阳性表达,正常皮肤表达均为阴性。iNOS在鳞状细胞癌、基底细胞癌及光线性角化病中的阳性率明显高于脂溢性角化病组(P<0.01),鳞状细胞癌组与光线性角化病组间无明显差别(P>0.05),与其他各组间差异显著(P<0.01或P<0.05),II III级鳞状细胞癌iNOS表达明显高于I级鳞状细胞癌(P<0.05)。结论:皮肤肿瘤中存在iNOS的表达,其合成的一氧化氮可能在皮肤的癌前病变及恶性肿瘤的发生、发展中起到一定的作用。其表达可能有助于皮肤肿瘤恶性度及预后的判断。     

Survivin和COX-2在皮肤基底细胞癌和鳞状细胞癌组织中的表达及相关性

目的了解皮肤基底细胞癌和皮肤鳞状细胞癌中Survivin和COX-2的表达情况及两者的关系。方法采用免疫组化法检测10例正常对照组、23例基底细胞癌、18例鳞状细胞癌组织中Survivin和COX-2的表达情况。结果 Survivin蛋白在正常组织中不表达,基底细胞癌和鳞状细胞癌中Survivin蛋白的表达率分别为60.87%和66.67%。COX-2在正常组织中的表达率为10%,基底细胞癌和鳞状细胞癌中COX-2的表达率分别为65.22%和66.67%,且明显高于其在正常组织中的表达率。Survivin的表达和COX-2的表达呈显著正相关(P0.05)。结论 Survivin蛋白和COX-2在皮肤基底细胞癌和皮肤鳞状细胞癌中高表达,两者呈正相关。     

周期素A在皮肤鳞状细胞癌与角化棘皮瘤中的检测

目的通过检测周期素(cyc lin)A在皮肤鳞状细胞癌(SCC)和角化棘皮瘤(KA)中的分布,探讨两者之间的关系。方法采用免疫组化技术检测12例KA和20例SCC皮损中cyc lin A蛋白。结果cyc lin A阳性细胞在KA中主要位于肿瘤的周边部,而在SCC中,呈弥漫性分布;cyc lin A在KA中的平均阳性率(13.2%)与高分化SCC(15.5%)差异无显著性(P>0.05),但与中、低分化SCC(23.4%,33.6%)差异有显著性(P<0.05)。结论cyc lin A在KA与SCC中的分布特点进一步说明了两者的异同,KA是否是SCC的分型尚需进一步探讨;cyc lin A在KA及各型SCC中的阳性率的不同说明cyc lin A可以作为评价皮肤肿瘤增殖程度的一个分子生物学指标。