恶性黑素瘤和普通痣细胞痣细胞外信号调节激酶途径相关蛋白的研究

目的 探讨恶性黑素瘤(MM)和普通痣细胞痣细胞外信号调节激酶(ERK)途径相关蛋白ERK1和ERK2以及细胞周期素D1(Cyclin D1)蛋白的表达情况,分析这3种蛋白与MM的临床病理关系。方法 采用免疫组化SP法对30例MM和13例普通痣细胞痣中ERK1、ERK2和Cyclin D1蛋白的表达进行检测,并进行临床病理相关性分析。结果 ERK1、ERK2和Cyclin D1在MM组织中表达较普通痣细胞痣组织明显升高(P<0.01),ERK1、ERK2和Cyclin D1与MM的Clark分级和Breslow厚度无明显相关性,ERK1和ERK2与Cyclin D1在MM组织中表达显著相关。结论 MM中ERK途径得到了过度激活,并可能促使Cyclin D1表达增加,因此它们在MM的发病机制中可能起到一定作用,而它们在普通痣细胞中仅在A型痣细胞表达增加。     

钠氢交换子调节因子-1及β-联蛋白在皮肤黑素瘤及痣细胞痣组织中的表达

目的 探讨皮肤黑素瘤及痣细胞痣组织中钠氢交换子调节因子-1 (NHERF1)和β-联蛋白表达的意义.方法 收集47例皮肤黑素瘤及37例痣细胞痣组织,用免疫组化方法对NHERF1和β-联蛋白在组织中的表达模式(膜表达、浆表达及核表达)和表达强度进行检测.结果 原位皮肤黑素瘤组织中NHERF1胞质中等/强阳性表达率为38％;而侵袭性皮肤黑素瘤组织中NHERF1胞质中等/强阳性表达率为71％,两组相比差异有统计学意义.同时,全部皮肤黑素瘤组织中NHERF1胞质中等/强阳性表达率为60％,而痣细胞痣组织中NHERF1胞质中等/强阳性表达率为0,差异有统计学意义.原位皮肤黑素瘤组织中可见β-联蛋白胞质中等/强阳性表达率为50％;而侵袭性皮肤黑素瘤组织中可见β-联蛋白胞质中等/强阳性表达率为84％.两组相比差异有统计学意义.全部皮肤黑素瘤组织中B一联蛋白胞质中等/强阳性表达率为72％,而痣细胞痣组织中NHERF1胞质中等/强阳性表达率为30％,两组相比具异有统计学意义.NHERF1及β-联蛋白胞质中等/强阳性表达率在痣细胞痣组、原位黑素瘤组及侵袭性黑素瘤组中均随着病变组织的恶性程度增加而升高.运用Spearman秩相关检验对皮肤黑素瘤中NHERF1及β-联蛋白的表达相关性进行分析显示无相关性.结论 NHERF1及β-联蛋白在痣细胞痣、原位黑素瘤及侵袭性黑素瘤组织中的胞质阳性表达随着病变组织的恶性程度增加而升高,但两者的表达差异无相关性.     

38例色素痣免疫病理分析

目的:分析色素痣的免疫表型.方法:对38例色素痣进行免疫病理分析.结果:色素痣三种类型中S-100均呈强阳性表达;HMB45仅在真皮浅层阳性表达;Melan-A及层粘连蛋白在三种痣中表达强度和范围相同,但是强度比S-100表达弱,三者差别无统计学意义.增殖细胞核抗原(PCNA)在交界痣中表达明显增高.结论:PCNA在交界痣中活性增高.     

人类白细胞抗原-G在皮肤恶性黑素瘤中表达的研究

目的:检测人类白细胞抗原-G(HLA-G)在原发性皮肤恶性黑素瘤(CMM)中的表达情况.方法:以免疫组化法检测35例原发性CMM、22例痣细胞痣HLA-G及Bcl-2的表达,并结合临床及组织病理学进行分析.结果:在CMM中HLA-G蛋白的阳性表达率显著高于痣细胞痣(P=0.029).HLA-G蛋白表达与CMM炎细胞浸润、Bcl-2蛋白表达显著相关(P值分别为0.032,0.008).CMM中HLA-G表达与临床分期、肿瘤厚度、溃疡及生长模式等预后变量无显著相关性.结论:CMM中HLA-G表达上调,可能参与肿瘤的发生及发展.     

恶性黑素瘤和普通痣细胞痣组织中双链RNA依赖的蛋白激酶的表达

目的 探讨恶性黑素瘤和普通痣细胞痣组织中双链RNA依赖的蛋白激酶(PKR)的表达情况,并分析PKR与恶性黑素瘤的细胞增殖程度及临床病理参数的关系。方法 应用免疫组化法检测42例恶性黑素瘤及25例普通痣细胞痣组织中PKR及增殖细胞核抗原的表达。结果 恶性黑素瘤中PKR阳性表达率显著高于普通痣细胞痣组织。恶性黑素瘤中PKR的表达水平与增殖细胞核抗原的表达呈显著负相关;与患者性别、年龄、有无淋巴结转移及TNM分级等临床病理特征均无明显相关性。结论 PKR在恶性黑素瘤细胞增殖过程中具有重要作用。     

p16蛋白和PCNA在鳞状细胞癌组织中的表达及其与临床病理特征的关系

目的探讨p16蛋白和增殖细胞核抗原(PCNA)在鳞状细胞癌组织中的表达及其与临床病理特征的关系。方法利用免疫组织化学法(IHC)检测50例鳞状细胞癌患者的癌组织、癌旁组织和30例健康人群的正常组织中p16蛋白和PCNA的表达情况,并分析p16蛋白和PCNA与鳞状细胞癌临床病理特征的关系。结果鳞状细胞癌组织中p16蛋白阳性表达率显著低于癌旁组织和正常组织(P0.01),鳞状细胞癌组织中PCNA指数显著高于癌旁组织和正常组织(P0.01);p16蛋白阳性表达率随着癌组织浸润深度增加而降低(P0.01),PCNA指数随着癌组织浸润深度增加而增大(P0.01);PTNMⅠ~Ⅱ期p16蛋白阳性表达率显著高于PTNMⅢ~Ⅳ期(P0.01),PTNMⅠ~Ⅱ期PCNA指数显著低于PTNMⅢ~Ⅳ期(P0.01);p16阳性表达的鳞状细胞癌组织中PCNA指数为30.67±9.82,显著低于p16阴性表达的鳞状细胞癌组织中的51.96±12.43(P0.01)。结论在鳞状细胞癌组织中p16蛋白缺失,而PCNA大量表达,且p16蛋白和PCNA表达与鳞状细胞癌的病理分期和浸润深度密切相关,提示p16蛋白和PCNA可作为评价鳞状细胞癌病变程度的重要指标,对指导治疗鳞状细胞癌有重大意义。     

端粒酶催化亚单位蛋白及PCNA在尖锐湿疣组织中的表达及意义

目的探讨尖锐湿疣(CA)组织中端粒酶催化亚单位蛋白(hTERT)和增殖细胞核抗原(PCNA)的表达及意义。方法用免疫组化SP法分别检测35例CA组织和18例正常上皮组织中hTERT和PCNA的表达。结果hTERT蛋白在CA组织中阳性29例,阳性率为82.86%,在18例正常上皮中,阳性1例,阳性率为5.56%,二者比较差异有显著性(P<0.01);PCNA在35例CA组织中阳性30例,阳性率85.71%,而18例正常上皮中PCNA阳性率55.56%,PCNA两组阳性率之间差异有显著性(P<0.05);hTERT和PCNA之间的阳性表达呈正相关(r=0.681,P<0.01)。结论hTERT在CA发生、发展中起一定作用,hTERT和PCNA表达呈正相关,表明hTERT与细胞的增殖有关。     

MMP-2、MMP-9在恶性黑色素瘤中的表达及预后意义

目的观察基质金属蛋白酶(MMP)-2、MMP-9在恶性黑色素瘤(Malignant melanoma,MM)细胞中的表达以及预后意义。方法采用免疫组化SP法分别检测52例MM组织、20例色素痣中MMP-2、MMP-9的表达,用χ^2检验分析MM组织与色素痣组织中二者表达的差异性,用多变量Cox回归分析其预后关系,单因素Kaplan-Meier法和对数秩检验进行生存分析。结果MM组织中MMP-2(90.4%)、MMP-9(78.8%)表达均显著高于色素痣组织(P<0.05);MMP-2、MMP-9表达阳性患者死亡风险显著升高;MMP-2阳性表达的MM患者的生存期显著低于阴性者(均P=0.004),MMP-9阳性表达的MM患者的生存期显著低于阴性者(均P<0.001)。结论MMP-2及MMP-9在MM组织中高表达,且与患者预后关系密切。     

PCNA、P21和COX-2在尖锐湿疣中的表达及其意义

目的：研究尖锐湿疣组织中增殖细胞核抗原（PCNA）、P21及环氧化酶-2（COX-2）蛋白表达及其意义。方法：应用免疫组化SP法对30例尖锐湿疣组织及10例正常组织中PCNA、P21及COX-2蛋白进行检测。结果：30例尖锐湿疣组织中PCNA、P21阳性表达较正常对照组有不同程度的增加，有统计学意义；COX-2蛋白在尖锐湿疣组织中阳性表达率13．3％（4／30），正常组织无阳性表达（0／10），两者之间无显著性差异（P〉0．05）；尖锐湿疣组织中PCNA与P21表达之间无相关性（r=0．196，P〉0．05）。结论：PCNA、P21过表达与尖锐湿疣角质形成细胞自限性增殖密切相关。     

Bmi-1和PCNA在皮肤鳞状细胞癌组织中的表达及意义

目的检测Bmi-1和PCNA在皮肤鳞状细胞癌组织中的表达,并探讨Bmi-1表达与皮肤鳞状细胞癌临床病理特征的关系。方法采用免疫组化SP法检测Bmi-1和PCNA在30例皮肤鳞状细胞癌组织和30例脂溢性角化症皮损中的表达,并分析Bmi-1的表达与皮肤鳞状细胞癌临床病理特征的相关性。结果 Bmi-1在皮肤鳞状细胞癌和脂溢性角化症中的阳性表达率分别是70.00%和16.67%,前者显著高于后者(P<0.001),而且Bmi-1的表达与病理分级、病理分期和淋巴结转移相关(P<0.05)。PCNA的阳性表达率分别是96.67%和66.67%,前者也显著高于后者(P<0.01)。但Bmi-1蛋白在皮肤鳞状细胞癌中的表达和PCNA的表达无相关关系(P>0.05)。结论 Bmi-1和PCNA在皮肤鳞状细胞癌组织中高表达,而且Bmi-1与皮肤鳞状细胞癌的分化、浸润和转移可能相关。     

生存素在恶性黑素瘤中的表达及与VEGF和PCNA的关系

目的 探讨人恶性黑素瘤组织中凋亡抑制蛋白生存素的表达,及其与恶性黑素瘤临床特征、血管内皮生长因子(VEGF)和增殖细胞核抗原(PCNA)表达的关系.方法 免疫组化方法检测36例人恶性黑素瘤和30例色素痣组织中生存素、VEGF和PCNA蛋白表达.结果 ①生存素在恶性黑素瘤和色素痣中均有表达,但在恶性黑素瘤中表达明显高于色素痣(P<0.01).②在恶性黑素瘤组织中有30例表达VEGF(阳性率83.3%),36例表达PCNA(阳性率100%),而色素痣组织不表达VEGF和PCNA.③生存素的阳性表达与恶性黑素瘤患者的发病年龄、性别、淋巴结转移均无相关性(P>0.05),但其表达与VEGF和PCNA的表达密切相关(P<0.01).结论 生存素可能参与恶性黑素瘤的增殖和与VEGF相关的肿瘤血管生成.     

VEGF-C及其受体在皮肤恶性黑素瘤中的表达及其临床意义

目的：研究皮肤恶性黑素瘤（MM）组织中血管内皮生长因子C（vascular endothelial growth factor-C,VEGF-C）及其受体VEGFR-3（flt-4）的表达情况,分析其在肿瘤淋巴管的生长及淋巴转移中的作用。方法：采用免疫组化SP法分析29例MM石蜡标本中VEGF-C/flt-4蛋白的表达情况,并与20例色素痣石蜡标本中VEGF-C/flt-4蛋白表达情况相对比,统计分析其与临床病理特点之间的关系。结果：VEGF-C蛋白的阳性率为96.6%（28/29）,flt-4的阳性率为86.2%（25/29）,明显高于色素痣（P〈0.01）,与肿瘤的病理分级、临床分期和肿瘤的淋巴转移显著相关（P〈0.05）;VEGF-C和flt-4的阳性表达相一致。结论：VEGF-C/flt-4在皮肤恶性黑素瘤中的表达明显高于色素痣,与肿瘤的预后密切相关,在肿瘤淋巴转移中可能有重要作用。     

Expression of endothelial nitric oxide synthase and vascular endothelial growth factor in human malignant melanoma and their relation to angiogenesis

BACKGROUND: Angiogenesis is the major and key factor for growth and invasion of tumours, including malignant melanoma (MM), but the factors that contribute to tumour angiogenesis are still unclear. OBJECTIVE: To study expression of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) in human MM and their relation to angiogenesis. To investigate the correlation between eNOS and VEGF and the role of nitric oxide (NO) generated by eNOS in the process of mediating angiogenesis by VEGF. METHODS: Tissue sections from 31 patients with MM were examined using immunohistochemistry and morphological quantitative analysis for protein expression of eNOS and VEGF. Microvessel density (MVD) was counted in endothelial cells in immunostained by anti-FVIII:RAg antibody. RESULTS: Positive eNOS and VEGF immunostaining were observed in 77.4% and 83.9% of MM lesions, respectively, whereas pigmented naevi never expressed eNOS and VEGF. A positive correlation between eNOS and VEGF in MM was observed. Expression of eNOS and VEGF was positively correlated with MVD expression in MM, and MVD expression in MM was stronger than in pigmented naevi. Expression of eNOS and VEGF was not correlated with lymph node metastasis. CONCLUSIONS. On the basis of the current data showing that malignant melanocytic tumours displayed strong VEGF and eNOS expression, whereas benign melanocytic proliferations showed no immunoreactivity for VEGF and eNOS, such expression may be used as a discriminating factor to distinguish malignant melanoma from pigmented naevi. Expression of eNOS and VEGF may contribute to angiogenesis of MM, eNOS probably plays an important role in mediating VEGF-induced angiogenesis.     

恶性黑素瘤血管内皮细胞生长因子和nm23抑癌基因蛋白表达及微血管密度的研究

目的 探讨血管内皮细胞生长因子(VEGF)、nm23抑癌基因和微血管密度(MVD)与恶性黑素瘤发病的关系。方法 采用免疫组化法检测47例恶性黑素瘤和10例黑素细胞痣组织中VEGF、nm23和CD34的表达并计算其MVD。结果 恶性黑素瘤VEGF阳性表达率和MVD值均明显高于黑素细胞痣(P均<0.01),有淋巴结转移者明显高于无淋巴结转移者(P值分别<0.05和<0.01);随着浸润深度增加,Ⅰ级与Ⅳ、Ⅴ级恶性黑素瘤比较,VEGF阳性表达率和MVD值差异均有统计学意义(P值分别<0.05和<0.01)。恶性黑素瘤中nm23阳性表达率明显低于黑素细胞痣(P<0.01),有淋巴结转移者明显低于无淋巴结转移者(P<0.01),与临床分型和浸润深度无明显关系。结论 VEGF和nm23表达在恶性黑素瘤的发展和转移中起重要作用。     

孕激素对无经典孕激素核受体黑素瘤细胞体外生长的影响

目的研究孕激素对黑素瘤体外生长的影响及信号传导机制。方法以间接免疫荧光法检测黑素瘤细胞系A375及A875孕激素受体表达;MTT法及流式细胞术分别检测细胞增殖与凋亡,Western blot法检测ERK1/2磷酸化水平。结果A375及A875均无细胞核内孕激素受体蛋白表达;低浓度孕激素对A375及A875表现为促生长效应,随浓度增高促增殖效应减弱并出现抑制效应;低浓度促生长效应可被MEK1/2抑制剂U0126抑制,但不受孕激素受体拮抗剂RU486影响;高孕激素浓度促进细胞凋亡;低浓度孕激素上调ERK1/2磷酸化水平,高浓度作用相反。结论孕激素可通过不依赖经典细胞核受体的非基因组机制调节黑素瘤细胞的生长及凋亡。     

恶性黑素瘤内皮型一氧化氮合酶和血管内皮生长因子的表达

目的 探讨内皮型一氧化氮合酶(eNOS)和血管内皮生长因子(VEGF)与人恶性黑素瘤血管生成的关系。方法 免疫组化法检测31例人恶性黑素瘤标本eNOS、VEGF和第Ⅷ因子相关抗原(FⅧRAg)血管内皮细胞特异性染色,计数肿瘤微血管密度(MVD)。结果 24例恶性黑素瘤组织表达eNOS,26例表达VEGF,色素痣组织不表达eNOS和VEGF;VEGF与eNOS的表达呈正相关;eNOS、VEGF的表达与恶性黑素瘤MVD呈正相关,恶性黑素瘤MVD明显高于色素痣;eNOS、VEGF表达与淋巴结转移无关。结论 MVD随着eNOS和VEGF表达的增强而增加。     

Oestrogen receptor beta and melanoma: a comparative study

Background Oncological research has focused on evaluating oestrogen receptors (ERs) in oestrogen‐related tumours, and understanding the potential role of ERs in the pathophysiology of cancer. Objectives To investigate the significance of oestrogen receptor beta (ERβ) in melanoma. Methods We prospectively evaluated ERβ expression in malignant melanoma (MM) tissue and adjacent healthy skin by quantitative immunohistochemistry at the Department of Dermatology of the University of Florence, from 1998 to 2010. Results ERβ was detected with varying staining intensity in the 66 malignant melanocytic lesions. After adjusting for age and sex, we found that ERβ expression was significantly lower in melanoma tissue compared with adjacent healthy skin (P < 0·0001). We also found significantly lower ERβ levels in thick melanoma tissue compared with thin melanoma tissue. In addition, there was a positive association between Breslow thickness and the difference of ERβ expression between healthy tissue and melanoma tissue (P = 0·0004). Consistent with sex differences in melanoma survival, men showed significantly lower levels of ERβ than women in both melanoma (P = 0·05) and healthy tissues (P = 0·02). Conclusions ERβ expression is inversely associated with Breslow thickness and is significantly influenced by sex in MM.     

Proliferative activity of cutaneous melanocytic neoplasms defined by a proliferating cell nuclear antigen labelling index

Summary To evaluate the proliferative activity of benign, borderline and malignant cutaneous melanocytic neoplasms, 30 cases of malignant melanoma (MM) and 41 cases of naevi were studied by immunostaining using a monoclonal antibody against proliferating cell nuclear antigen (PCNA). PCNA is a nuclear antigen expressed in the late G1 and S phase and serves as a marker of proliferating cells. Invasive MM and MM in situ showed much higher PCNA positivity rates than melanocytic naevi (invasive MM, 18.0%; MM in situ, 11.3%; ordinary melanocytic naevi, 2.6%). The PCNA positivity rate did not increase significantly with the thickness of MM. Among ordinary melanocytic naevi, junctional naevi had a higher PCNA positivity rate than compound or intradermal naevi. Mean PCNA positivity rates for Spitz's naevi and sporadic dysplastic naevi were within the range for ordinary melanocytic naevi, indicating the benign nature of both types of naevus. Contrary to some previous studies, MM in situ showed high proliferative activity, indicating that cells of MM in situ are actively proliferating. This study clearly demonstrates that MM and various types of naevi can be separated according to differences in proliferative activity defined by the PCNA labeling index.This work was presented in part at the 14th International Pigment Cell Conference, Kobe, Japan, October 31–November 4, 1990     

Oestrogen and progesterone receptors in melanoma and nevi: an immunohistochemical study

Background

The effect of hormonal stimulation and fertility treatments, on the development of malignant melanoma (MM) remains to be determined.

Objectives

The aim of this study was to investigate the presence of oestrogen receptor alpha (ERα) and progesterone receptor (PR) inMM and nevi after hormonal stimulation.

Materials & Methods

Immunohistochemical analyses were performed utilizing antibodies specifically directed against ERα and PR in MM and atypical nevi specimens from patients: (1) diagnosed during pregnancy, (2) diagnosed in the six months following delivery, or (3) who had undergone repetitive cycles of hormonal stimulation for in vitro fertilization (IVF) in the year that preceded MM diagnosis. Controls were atypical nevi and MM specimens of female patients of the same age group who had received no hormonal therapies and reported no pregnancies in the five years before diagnosis.

Results

Twenty-eight female patients at childbearing age were selected for this study. Strong cytoplasmic positivity of ERα and PRwas detected in atypical melanocytes of two MM specimens of patients who had undergone repetitive cycles of hormonal stimulation during IVF procedures. All other specimens showed no expression ofERαor PR.

Conclusion

Since our results represent preliminary findings, conclusions regarding a possible correlation between IVF therapy and melanoma occurrence cannot be ascertained. Larger laboratory studies should be performed to investigate reproductive hormone receptor expression in MM in women following IVF, pregnancy, prolonged contraceptive use, or hormone replacement therapy.

     

Immunohistochemical expression of cyclooxygenage-2 in melanocytic skin lesions

Background Several reports have shown expression of cyclooxygenase‐2 (COX‐2) in malignant skin tumors. COX‐2 has also recently been reported as a marker of malignant melanoma (MM). Objective Our aim was to investigate whether there is a difference in the immunohistochemical expression of COX‐2 between malignant and benign melanocytic lesions of the skin. Methods We selected 40 archival cases of MM including 10 cases of superficial spreading melanoma, 10 of lentigo maligna melanoma, 10 of nodular melanoma, and 10 of acral lentiginous melanoma. For comparison, we also selected 35 benign melanocytic lesions, which included 15 nonatypical nevi and 10 atypical nevi. The remaining 10 cases were Spitz nevi. COX‐2 immunohistochemical staining was performed, and intensities were assessed quantitatively. Results The MM group and the benign melanocytic nevi group showed a highly statistically significant difference in the intensity of COX‐2 expression (P < 0.0001). Staining intensity in the dermal component of MM cases also showed a tendency to increase with increasing tumor depth. By contrast, the intensity of the dermal component in the melanocytic nevi group decreased with increasing depth as the nevus cells matured from type A to type C cells. No statistical difference was noted between the MM and Spitz nevi cases (P = 0.20). Conclusions Malignant melanoma shows stronger immunohistochemical expression of COX‐2 than benign melanocytic nevi. Although COX‐2 cannot be used alone to differentiate MM from melanocytic nevi, it may serve as an aid in the differential diagnosis of melanocytic skin lesions.