皮肤恶性肿瘤的DNA异常甲基化研究进展

对肿瘤研究的不断深入发现除了基因突变之外,表观遗传改变也与肿瘤的发生发展密切相关。肿瘤表观遗传的改变以DNA的异常甲基化为主,其主要的机制是由肿瘤抑制基因启动子超甲基化引起基因沉默。DNA的异常甲基化几乎存在于任何类型的人类肿瘤中,包括皮肤恶性肿瘤。现对DNA甲基化、DNA甲基化与肿瘤、DNA甲基化在基底细胞癌、鳞状细胞癌、黑色素瘤和皮肤淋巴瘤等皮肤恶性肿瘤中的研究进行综述。     

微小核糖核酸在皮肤肿瘤的研究进展

已有研究证实,miRNA参与恶性黑素瘤的发生,是诊断和治疗的新靶目标.近年研究发现,miRNA的调节异常已经成为基底细胞癌、皮肤鳞状细胞癌发病机制的一条重要途径;而且miRNA在其他皮肤肿瘤(蕈样肉芽肿、Merkel细胞癌)组织中的表达水平有不同程度的上调或下调,是这些肿瘤发生发展的影响因素.病毒编码的miRNA调节病毒基因的表达,参与病毒相关性皮肤肿瘤的发生.     

皮肤基底细胞癌T钙黏蛋白的表达及其DNA异常甲基化分析

目的 检测皮肤基底细胞癌(BCC)中T钙黏蛋白的表达以及异常甲基化.方法 采用免疫组化法和CpG岛甲基化特异性PCR方法,对29例BCC和19例正常人暴光部位皮肤的组织标本进行T钙黏蛋白的表达及甲基化检测.结果 正常皮肤皆表达T钙黏蛋白,29例BCC中8例(27.59%)检测到T钙黏蛋白表达(P<0.05),9例BCC(31.03%)和19例正常皮肤中的1例(5.26%)检测到T钙黏蛋白启动子区域的异常甲基化(P<0.05).结论 T钙黏蛋白表达降低与皮肤基底细胞癌的发病机制相关,T钙黏蛋白启动子区域的异常甲基化与蛋白低表达可能相关.     

炎症性皮肤病的DNA异常甲基化

DNA甲基化是一种重要的不改变DNA序列,但可影响基因表达水平并遗传的表观遗传学事件.随着DNA甲基化检测方法的日趋完善,有越来越多的研究表明,DNA异常甲基化与炎症性皮肤病相关.有研究证实或提示,DNA异常甲基化可能与多种炎症性皮肤疾病的发病机制密切相关,如银屑病、系统性红斑狼疮、特应性皮炎、系统性硬皮病,其他尚包括雄激素性脱发、慢性湿疹和天疱疮等,但研究并不能清楚地解释异常的甲基化模式如何在疾病中发挥具体的生物学作用.随着研究的深入,可对相关疾病的研究提供新的思路.     

皮肤肿瘤

20110284皮肤鳞状细胞癌中FHIT基因启动子甲基化与FHIT蛋白表达的临床意义/田中伟(新乡医学院一附院皮肤科),宋向凤,李敏…∥现代预防医学.-2010,37(6).-1176~1182采用甲基化特异性PCR法测定32例皮肤鳞状细胞癌组织标本FHIT基因启动子甲基化情况,免疫组化法检测其蛋白表达情况。结果有17例发生了甲基化,阳性率53.10%;皮肤鳞状细胞癌及正常皮肤组织中     

E-钙黏蛋白与皮肤鳞状细胞癌

E-钙黏着蛋白是介导上皮细胞间粘连的最重要的一种钙依赖性的细胞黏附分子.近年来研究发现E-钙黏着蛋白异常表达与多种上皮恶性肿瘤的分化、浸润生长、转移和预后有着密切的关系.研究E-钙黏着蛋白的结构功能、在肿瘤侵袭转移中的作用机制、与皮肤鳞状细胞癌的相关性及临床意义,以期对皮肤鳞状细胞癌的早期诊断、治疗选择和预后预测提供一定依据.     

皮肤T细胞淋巴瘤表观遗传学研究进展

表观遗传学主要是研究在不改变DNA序列的情况下,发生的基因表达水平可遗传的变化,主要包括DNA甲基化、组蛋白修饰和非编码RNA调控.皮肤T细胞淋巴瘤是一组原发于皮肤的T淋巴细胞恶性增殖性疾病,蕈样肉芽肿和Sezary综合征是最常见的皮肤T细胞淋巴瘤.近年来研究表明,表观遗传学不仅在皮肤T细胞淋巴瘤的发生发展中起重要作用,而且可以通过改变表观遗传达到治疗皮肤T细胞淋巴瘤目的.     

皮肤鳞状细胞癌中p16基因甲基化的研究

近年来研究显示,p16基因第1外显予5'CpG岛的高甲基化在多种肿瘤的发生发展中起重要作用^[1],特别是在一些过去未发现有p16基因缺失与突变的肿瘤中也发现由p16基因甲基化而引起的基因失活。我们应用限制性内切酶-PCR法对40例原发性皮肤鳞状细胞癌(鳞癌)组织和10例正常皮肤组织进行pl6基因甲基化的检测,以探讨皮肤鳞癌的发生发展是否与pl6基因甲基化有关。     

皮肤鳞状细胞癌组织中p16及14-3-3σ基因启动子甲基化检测及其相关性分析

目的:通过检测皮肤鳞状细胞癌组织及正常皮肤组织中p16及14-3-3σ基因启动子甲基化,探讨p16及14-3-3σ基因启动子甲基化在皮肤鳞状细胞癌发生发展中的作用。方法:实时荧光定量聚合酶链反应(q-PCR)法检测53例皮肤鳞状细胞癌组织及48例正常皮肤组织中p16和14-3-3σm RNA表达,免疫印迹(western blot)法检测p16和14-3-3σ蛋白表达,应用甲基化特异性PCR对所提DNA进行p16及14-3-3σ基因启动子甲基化检测,并分析这两个基因的相关性。结果:皮肤鳞状细胞癌标本中p16及14-3-3σm RNA表达均低于正常皮肤组织,皮肤鳞状细胞癌标本中p16和14-3-3σ蛋白亦低于正常皮肤组织,皮肤鳞状细胞癌标本中p16及14-3-3σ基因启动子甲基化的阳性率分别为54.72%(29/53)和67.92%(36/53),高于对照组的8.33%(4/48)和18.75(9/48),差异有统计学意义(P0.001);皮肤鳞状细胞癌组织中p16和14-3-3σ的甲基化存在正相关关系(r=0.431,χ~2=9.825,P=0.002,P0.05)。结论:皮肤鳞状细胞癌标本中p16及14-3-3σ低表达及甲基化与鳞状细胞癌发生发展可能有关,皮肤鳞状细胞癌基因启动子甲基化可能是多位点参与的。     

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.     

转基因动物技术研究皮肤肿瘤

基底细胞癌和鳞状细胞癌是常见的皮肤肿瘤，其发病与多种因素有关。从遗传学的角度上看肿瘤是一种基因病，以往由于研究条件的限制，人们对于涉及的基因并不了解。随着转基因以及基因打靶技术的不断完善，研究者已经能够精确研究肿瘤发病中相关基因的具体作用，这对于明确肿瘤发病机制及指导治疗均提供有利的平台。以基底细胞癌和鳞状细胞癌为例，对转基因动物技术在皮肤肿瘤研究领域的最新研究进展概述。     

The relation between dermoscopy and histopathology of basal cell carcinoma

BACKGROUND:

Basal cell carcinoma is the most frequent cancer in fair-skinned populations and dermoscopy is an important, non-invasive technique that aids in the diagnosis of Basal cell carcinoma.

OBJECTIVES:

The aim of this study was to evaluate the relationship between histopathological subtypes and dermoscopic features of Basal cell carcinoma.

METHODS:

This study included 98 patients with clinically and histopathologically confirmed Basal cell carcinomas. The dermoscopic features of the lesions from each patient were analyzed before the histopathological findings were evaluated.

RESULTS:

Dermoscopic structures were observed in all 98 patients and irregular vascularity was identified in 78 patients (79.6%). The most common vascular pattern was the presence of arborizing vessels (42 patients, 42.9%) followed by arborizing microvessels (21 patients, 21.4%) and short fine telangiectasias (SFTs; 15 patients, 15.3%). White streaks (38 patients, 38.8%), translucency (31 patients, 31.6%), a milky-pink to red background (42 patients, 42.9%), and erosion/ulceration (29 patients, 29.6%) were also observed. Pigmented islands were seen as blue-gray globules (7 patients, 7.1%) and blue-gray ovoid nests (42 patients, 42.9%). The pigment distribution pattern was maple leaf-like areas in 9 patients (9.2 %) and spoke wheel-like areas in 6 patients (6.1%).

CONCLUSIONS:

Basal cell carcinomas show a wide spectrum of dermoscopic features. Arborizing vessels were the most common dermoscopic findings in Basal cell carcinomas, while superficial Basal cell carcinomas displayed mainly milky-pink to red areas, and arborizing microvessels. The most common dermoscopic features of pigmented types were islands of pigment (blue-gray globules, blue-gray ovoid nests). In conclusion, dermoscopy can be used as a valuable tool for the diagnosis of Basal cell carcinomas and prediction of their histopathological subtypes.     

Modified Tripier flap in reconstruction of the lower eyelid

We describe the use of modified Tripier flap for reconstruction of a surgical defect in the lower eyelid region, after excision of nodular basal cell carcinoma.     

皮肤鳞状细胞癌临床和病理分析

目的探讨皮肤鳞状细胞癌的临床和病理特点。方法收集2009--2012年经组织病理诊断为皮肤鳞状细胞癌的58例患者的临床及病理资料,对临床和组织病理学特征进行回顾分析。结果皮肤鳞状细胞癌高峰发病年龄为70～89岁,皮损见于头面部27例（46．55％）,外生殖器部位20例（34．48％）,四肢9例（15．52％）,躯干2例（3．45％）。临床上表现为结节隆起型27例,占46．55％,深在浸润型19例,占32．76％,溃疡型12例,占20．69％。组织病理表现为经典型51例,特殊类型7例（疣状型5例、腺性鳞癌1例、透明细胞型1例）。结论皮肤鳞状细胞癌好发于老年人曝光部位,应及早进行组织病理学检查以明确诊断,提高患者生存率。     

皮肤假血管肉瘤型鳞状细胞癌临床病理观察

目的 提高对易误诊为血管肉瘤的皮肤假血管肉瘤型鳞状细胞癌的认识.方法 报道2例皮肤原发的假血管肉瘤型鳞状细胞癌.结果 例1女性,71岁,皮损位于右小腿远端;例2男,57岁,皮损位于右颈部.皮损均为单发的溃疡性肿块.光镜下棘层显著松解形成假血管腔隙,内含游离肿瘤细胞和红细胞,腔隙内衬多角形或扁平、鞋钉样细胞.瘤细胞上皮样,异形性明显,核分裂象易见,胞质丰富,空泡状,核仁明显.肿瘤间质疏松,可见嗜碱性黏液样基质,局部区域弥漫出血坏死.免疫表型panCK、CK5/6、Vim、CK14、EMA、P63及P53阳性,血管内皮标记CD31、CD34、F8、Fli-1阴性.电镜下肿瘤胞质内存在少量的张力原纤维及典型的桥粒结构.结论 皮肤原发的假血管肉瘤型鳞状细胞癌是一种鳞状细胞癌亚型,需要与血管肉瘤、上皮样肉瘤等恶性肿瘤进行鉴别.     

Squamous cell cancer – 31-year epidemiological study in a city of south Brazil

BACKGROUND

The incidence and morbidity of squamous cell cancers are increasing worldwide. Epidemiological studies with morbidity coefficients about this type of cancer are scarce in Brazil.

OBJECTIVES

To determine morbidity coefficients, analyze and classify the squamous cell cancers diagnosed in the city of Blumenau - SC from 1980 to 2011, according to clinical and histological features.

METHODS

The authors revised 4000 histopathological exams with respect to sex, age, anatomic site and histological type. The morbidity coefficients were calculated using the number of squamous cell cancers found and the annual population estimated by the Brazilian Institute of Geography and Statistics between 1980 and 2011.

RESULTS

A total of 4000 tumors were identified, 2249 (56.2%) in male and 1751 (43.8%) in female patients. The standard incidence rates varied from 40 cases in 1980 to 120 cases/100,000 inhabitants in 2011. The morbidity above 70 years of age reached 1484 cases/100,000 inhabitants in male and 975 in female patients. As to primary anatomic site, we found more tumors on the lips and ears in male and on the face and legs in female patients. As to the degree of involvement, the more frequent were Well Differentiated Squamous cell carcinomas (70%) and Moderate Squamous cell carcinomas (19,1%). The Low Differentiated Squamous cell carcinomas, which represented those with the worst prognosis, were found in 4.5% of the tumors.

CONCLUSION

Squamous cell cancers in Blumenau - SC have similar patterns of distribution regarding age, primary anatomic site and histological types as found in the international literature. The morbidity increased by 300% in the last 31 years, which indicates that we need to dedicate special attention to the older population.     

人皮肤鳞状细胞癌A431细胞株电压门控钾通道的研究

目的 探讨电压门控钾通道在人皮肤鳞状细胞癌A431细胞和人角质形成细胞HaCaT细胞中的作用.方法 噻唑蓝方法研究加入含有不同浓度四乙胺的培养基对A431细胞和HaCaT细胞的增殖的影响;提取体外培养A431细胞、HaCaT细胞以及人正常表皮组织的蛋白,ELISA方法检测电压门控钾通道蛋白(HERG)在它们中的表达,比较它们的差异.结果 四乙胺对体外培养的A431细胞和HaCaT细胞的增殖抑制效应具有时间依赖与剂量依赖的特点,当四乙胺的浓度≥10 mmol/L且作用时间≥24 h,可以使A431细胞和HaCaT细胞的增殖受到明显的抑制.HERG钾通道蛋白在A431细胞、HaCaT细胞和人正常皮肤表皮组织均有表达,平均浓度分别为(49.7114±3.55696)pg/ml、(35.7471±4.14696)pg/ml、(36.8857±3.47810)pg/ml,组间比较差异具有统计学意义(P<0.05).结论 阻断电压门控钾通道可抑制A431细胞和HaCaT细胞的增殖;电压门控钾通道可能在人皮肤鳞状细胞癌细胞上的表达更显著.

Abstract：

Objective To investigate the role of voltage-gated potassium channel in the human skin squamous cell carcinoma A431 cells and human keratinocyte HaCaT cells. Methods MTT assay was performed to detect the effect of different concentrations of tetraethylammonium (TEA) on the proliferation of cultured A431 cells and HaCaT cells. Besides, enzyme-linked immunosorbent assay (ELISA) was conducted to detect the expression of HERG channel protein in A431 cells, HaCaT cells and normal human skin tissue.Results TEA inhibited the proliferation of A431 cells and HaCaT cells in a dose- and time-dependent manner.After treated with TEA (≥ 10 mmol/L) for 24 or more hours, the proliferation of A431 cells and HaCaT cells was obviously suppressed. A significant difference was observed in the average concentration of HERG channel protein between A431 cells, HaCaT cells and normal human skin tissue (49.7114 ± 3.55696 pg/ml, 35.7471 ±4.14696 pg/ml, 36.8857 ± 3.47810 pg/ml, all P < 0.05). Conclusions The block of voltage-gated potassium channel could inhibit the proliferation of A431 cells and HaCaT cells, and the expression of voltage-gated potassium channel seems to be higher in human skin squamous cell carcinoma cells.     

翻译控制肿瘤蛋白在鳞状细胞癌和细胞株A431及SCL-1中的表达

目的 探讨翻译控制肿瘤蛋白(TCTP)在人皮肤鳞状细胞癌(SCC)组织及其细胞株A431及SCL-1中的表达,观察其对SCC细胞凋亡及增殖的影响.方法 免疫组化法观察TCTP蛋白在SCC组织中的表达.Western印迹法检测TCTP蛋白在SCC细胞株A431及SCL-1中的表达情况.继而构建针对编码TCTP的基因TPT1的siRNA序列,通过脂质体转染法转入A431细胞中,RT-PCR法检测TPT1的表达,Western印迹法检测TCTP蛋白的表达,流式细胞仪检测细胞凋亡率,MTT法检测A431细胞增殖情况.结果 免疫组化结果显示,在SCC中存在着TCTP的高表达,且其表达水平与癌组织的分级有着正相关关系(P＜0.05).Western印迹结果证实,在A431细胞及SCL-1细胞株中都存在着TCTP的表达,以A431细胞表达为最高.荧光检测siRNA转染效率可达到90％ ～ 95％.RT-PCR及Western-Blot结果显示合成的siRNA可以有效下调A431细胞中TPT1及TCTP的表达(P＜0.05).结论 合成的siRNA可以有效下调A431细胞中TP及TCTP的表达,这种下调可引起A431细胞凋亡率的上升和细胞增殖的抑制.