恶性黑素瘤患者T细胞化学趋化功能

目的　探讨恶性黑素瘤患者循环T细胞在体外对单核细胞化学趋化蛋白 1 (MCP 1 )的化学趋化功能及其与肿瘤浸润淋巴细胞 (TIL)及与肿瘤转移之间的关系。方法　用微量化学趋化装置和免疫组化方法测定患者T细胞对MCP 1的化学趋化功能和TIL计数。结果　原发性和转移性黑素瘤患者T细胞对MCP 1的化学趋化功能均明显低下 ,而基底细胞癌患者T细胞的化学趋化功能正常。免疫组化研究发现原发性恶性黑素瘤TIL数量与T细胞的化学趋化功能之间无关。结论　原发性和转移性恶性黑素瘤患者循环T细胞特异性地对MCP 1的化学趋化反应低下。这可能是由于这些细胞上的MCP 1受体表达或调节异常所致     

CD99 Immunoreactivity in Metastatic Malignant Melanoma

CD99, also known as p30/32, is a glycoprotein product of the MIC2 gene, which is located on the short arm of both chromosome X and Y. This transmembrane protein was originally utilized in immunohistochemistry as a unique marker for Ewing sarcoma, other primitive neuroectodermal tumors, and more recently in a wide variety of tumors. It's expression in malignant melanoma (MM) has not been well documented. A recent study at our institution demonstrated membranous staining in approximately 61% of primary MM. As CD99 is expressed by hematopoeitic cells, it has been proposed as a mechanism for lymphocytes to gain access to the vasculature.1 This study is designed to determine if CD99 expression in melanoma cells has a similar role using cases of metastatic MM from our archives. Our evaluation shows that 13 of 28 cases (46.4%) demonstrated membranous CD99 staining. A case of this magnitude has not been previously reported. Reference: 1. Shenkel AR, Mamdouh Z, Chen X, Liebman RM, Muller WA. CD99 plays a major role in the migration of monocytes through endothelial junctions. Nature Immunol 2002;3:143–150.     

Balloon Cell Melanoma: Case Report

Texts and atlases have limitations when used to teach dermatopathology to trainees: they can be tedious and time‐consuming to use, image detail is limited, text and figure legends can be ambiguous, and only a minor degree of image annotation (letters, arrows, lines outlining important features) is possible. A video screen recording software program (Camtasia Studio) has been used to create high quality digital audio and video dermatopathology tutorials. The screen recording program is started with the image window being recorded. Audio narration (USB digital microphone) is recorded along with live drawings of annotating lines outlining important histologic features, creating the effect of a "digital chalkboard" being used by the instructor. Separate clips of each "chalkboard annotation" with accompanying narration can be combined to create a seamless movie detailing each of the important histopathologic features leading to a specific dermatopathologic diagnosis. Video dermatopathology tutorials are extremely effective because they closely mimic instruction at the microscope. However, they offer the significant advantage of allowing remote teaching at any time or place convenient for the learner. Tutorials can be compressed into a variety of multimedia formats and streamed for web delivery or can be provided on CD for distribution.     

皮肤转移性黑素瘤1例

患者男,53岁。左大腿肿块伴瘙痒2月余。结合病史、皮损特点、实验室检查、皮损组织病理学改变、免疫组化检查结果,确诊为皮肤转移性黑素瘤。     

肿瘤干细胞及黑素瘤干细胞的研究进展

干细胞（stem cell，SC）是一类具有自我更新和多向分化潜能的原始细胞，SC可用于研究增殖、分化、生长、发育、衰老、死亡等最基本的生命现象，还可经体外诱导，用于临床细胞治疗和组织工程材料，为一些目前难治性疾病的治疗带来新的希望。随着对干细胞生理特性认识的深入，发现干细胞与肿瘤细胞有很多共同特点，认为肿瘤很可能是干细胞在长期的自我更新过程中，由于多基因突变导致干细胞生长失去正常调控，而停止在分化的某一阶段无限制增殖所形成的结果，由此提出肿瘤的肿瘤干细胞（cancerstem cells，CSC）学说。目前已在白血病、乳腺癌、脑胶质瘤、黑素瘤等肿瘤中分离出CSC并深入研究了CSC的生物学特性，为肿瘤研究开辟了新的途径，是人类认识肿瘤的一大进步，本文就CSC及黑素瘤干细胞研究的进展做一综述。     

Giant Metastatic Malignant Melanoma with an Unknown Primary Site

We report a case of malignant melanoma of unknown primary origin which presented with a giant metastatic tumor in his right inguinal region. A 94-year-old man noticed a small subcutaneous tumor in the right inguinal region 3 years earlier, which eventually became as large as 9 cm in diameter without treatment. Although a histological examination of the lesion showed malignant melanoma, extensive examination did not reveal its primary lesion or any metastasis other than that to the right inguinal area. Our case took an interesting course in that this well-growing metastatic tumor was localized in only one region and supported a previous report indicating that malignant melanoma with unknown primary origin has a low tendency to metastasize and a relatively good prognosis.     

A Case of Malignant Melanoma with Multiple Primary Lesions

Merkel cell carcinoma (MCC) is a small round cell tumor which does not characteristically stain with S‐100 protein. We report a case of an 83 year‐old male with a two‐month history of an ulcerated nodule on the right arm. Histopathology revealed small blue round cells arrayed in a trabecular pattern and in diffuse sheets, with scattered mitoses and necrosis. The cells were negative for vimentin, positive for NSE, focally positive for high molecular weight cytokeratins and diffusely positive for low molecular weight cytokeratins, including CK20. The findings were diagnostic of MCC, however, foci of S‐100 protein positive cells were also present in the tumor. Upon reviewing cases from Skin Pathology Laboratory at BUSM received between 1/1/2000 to 7/30/2004, we documented a total of 34 cases diagnosed as MCC; 24 were stained with S‐100 protein and this case was the only one that stained positively. S‐100 protein positivity of MCC, while having been previously reported, is an uncommon finding. We report our case to stress the importance of morphological and immunoperioxidase features for the diagnosis of Merkel cell carcinoma, particularly in the exceptional cases which exhibit S‐100 protein positivity.     

乙酰肝素酶mRNA和蛋白在恶性黑素瘤皮损及A375细胞株中的表达

目的探讨乙酰肝素酶mRNA和蛋白在恶性黑素瘤皮损及A375细胞株中的表达及意义。方法选择恶性黑素瘤30例、黑素细胞痣30例、恶性黑素瘤A375细胞株及15例正常皮肤组织,分别采用原位杂交、免疫组化法检测皮损及细胞株中乙酰肝素酶mRNA及蛋白的表达。结果A375细胞株及19例(19/30,63.33%)恶性黑素瘤皮损中乙酰肝素酶mRNA呈阳性表达,2例(2/30,6.67%)黑素细胞痣皮损中乙酰肝素酶mRNA表达阳性,正常皮肤组织中无1例呈阳性表达;恶性黑素瘤皮损中乙酰肝素酶蛋白的阳性表达率与黑素细胞痣、与正常对照组相比,差异均有显著性意义(χ12=19.200,P=0.000;χ22=25.714,P=0.000)。乙酰肝素酶蛋白在恶性黑素瘤、恶性黑素瘤A375细胞株、黑素细胞痣及正常皮肤组织的表达情况与mRNA相一致。结论乙酰肝素酶mRNA和蛋白的高表达可能与恶性黑素瘤的发生发展有关,有望作为治疗黑素瘤的靶点。     

全身转移性黑素瘤1例

患者女,60岁。头面、躯干、四肢大小不等黑红色结节,无痛痒3年,半年内泛发全身。皮肤科情况:头面、躯干、四肢见大小不等高粱粒至黄豆大小结节,呈皮色、暗红色及黑色。左膝关节上方见一直径约2cm×2cm大小肿块,其上见4～5个大小不等皮色至黑色结节,部分结节表面结痂。皮损组织病理示:真皮内弥漫性肿瘤样细胞浸润,此肿瘤样细胞核大、核仁明显,部分肿瘤样细胞呈巢状排列,肿瘤组织内见坏死。免疫组织化学染色显示肿瘤细胞LCA(-),CK20(-),S100蛋白及HMB45弥漫强阳性,Vimentin弥漫阳性。诊断:黑素瘤。     

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。     

芹菜素对人恶性黑素瘤A375细胞周期的影响及机制研究

目的研究芹菜素对人恶性黑素瘤细胞系A375细胞周期的影响,并进一步探讨其作用机制。方法 A375细胞经芹菜素干预后,MTT比色法检测芹菜素对细胞增殖的影响,PI单标流式细胞术检测芹菜素对细胞周期的影响,蛋白印迹法检测芹菜素对细胞周期相关蛋白Cyclin D1及p21表达的影响。结果芹菜素在体外能明显抑制A375细胞增殖,其抑制效应呈时间及浓度依赖性;芹菜素可影响A375细胞周期,经芹菜素干预的A375细胞,其细胞周期被阻滞于G1期;芹菜素可显著影响A375细胞周期相关蛋白的表达,芹菜素在下调A375细胞Cyclin D1表达的同时,明显上调p21蛋白的表达。结论芹菜素能拮抗人恶性黑素瘤A375细胞增殖,其机制可能与芹菜素影响A375细胞周期相关蛋白Cyclin D1/p21的表达进而调控A375细胞周期进程有关。     

Acute Kidney Injury in a Patient with Melanuria, Diffuse Melanosis, and Metastatic Malignant Melanoma

Diffuse melanosis with melanuria is a rare complication of metastatic malignant melanoma that is associated with a very poor prognosis. Acute kidney injury secondary to melanuria has not been reported previously. We describe a patient with malignant melanoma, diffuse melanosis, and melanuria who developed acute kidney injury that was clinically indicated by an increase in urinary excreted metabolites in the serum. Histopathologic examination of kidney tissue revealed damage to tubules associated with disseminated intra-tubular melanin deposits. Diffuse melanosis with melanuria may lead to morphologic and functional kidney changes and warrants careful monitoring of kidney function in affected malignant melanoma patients.     

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。     

Prognostic Significance of Epidermal Growth Factor Receptor Expression in Distant Metastatic Melanoma from Primary Cutaneous Melanoma

BackgroundEpidermal growth factor receptor (EGFR) is overexpressed in many cancers. However, EGFR expression in melanoma and its role are conflicting.ObjectiveThis study aimed to evaluate EGFR expression in distant metastatic melanoma and analyze its relationship with histologic and clinical characteristics and survival.MethodsDiagnostic tissues from 55 cases of distant metastatic melanoma was evaluated by immunohistochemistry for EGFR expression. Clinicopathologic features and survival outcomes were analyzed according to EGFR expression.ResultsThe positive EGFR expression in distant metastatic melanoma was significantly correlated with the absence of ulceration. The EGFR expression in distant metastatic melanoma was significantly associated with poor survival, under the conditions of male sex and primary cutaneous melanoma without ulceration or Breslow thickness ≤4.0 mm. This study bears limitations of a retrospective study in a single institution.ConclusionEGFR immunostaining had predictive values for survival outcome. The EGFR expression in distant metastatic melanoma in male, no ulcer, or Breslow thickness ≤4.0 mm appeared to be involved in disease progression.     

TRP-1编码基因反义核酸对良恶性黑素细胞凋亡的影响

目的　观察TRP 1反义核酸对黑素细胞及恶性黑素瘤细胞凋亡的影响。方法　将TRP 1反义核酸真核表达载体转染至黑素细胞及恶性黑素瘤细胞 ,以透射电镜及流式细胞仪分别观察细胞的凋亡情况。结果　电镜下可见各期凋亡细胞及凋亡小体 ;流式细胞仪检测表明TRP 1反义核酸转染黑素细胞及空载体转染对照组黑素细胞的凋亡率分别为 2 4.4%及 1.2 % ,TRP 1反义核酸转染的恶性黑素瘤细胞及空载体转染对照组恶性黑素瘤细胞的凋亡率分别为7.0 %及 2 .2 %。结论　TRP 1基因在维持良、恶性黑素细胞生长方面发挥重要作用 ,其反义核酸能明显促进良恶性黑素细胞的凋亡。     

恶性黑素瘤和良性黑素细胞肿瘤bcl-2蛋白表达

目的探讨ｂｃｌ ２蛋白在恶性黑素瘤（ＭＭ）中的表达及其意义。方法应用免疫组化方法对２４例ＭＭ的ｂｃｌ ２蛋白表达进行了检测。结果１５／２４的ＭＭ的瘤细胞阳性,且ｂｃｌ ２蛋白表达强度与ＭＭ的病理分级呈低度正相关（ｒｓ′＝０．４７１,Ｐ＜０．０５）,而良性黑素细胞肿瘤则极少表达ｂｃｌ ２蛋白。结论ｂｃｌ ２蛋白与ＭＭ的发展有关。