PDCD5及P53蛋白在不同期蕈样肉芽肿患者皮损中的表达

发布时间：2018-06-15 19:37

本文选题：程序性细胞凋亡因子5 + 蕈样肉芽肿　；参考：《山东大学》2011年硕士论文

【摘要】：研究背景：蕈样肉芽肿,又名蕈样霉菌病(mycosis fungoides, MF)是起源于记忆性辅助性T细胞的低度恶性皮肤T细胞淋巴瘤,临床可分为红斑、斑块、肿瘤三期,病程呈慢性进行性,晚期可以侵犯全身各个脏器。MF的发病机制目前尚未完全明确,近年来对MF中树突状细胞的分布及功能、皮肤T淋巴细胞的归巢机制、T淋巴细胞凋亡障碍及凋亡相关基因如p53、p16、bcl-2等异常表达的研究,为MF的发病机制及治疗提供了一些新的思路。亲表皮现象是MF组织病理学的特征之一,这提示表皮细胞与淋巴细胞的相互作用可能在MF的发病中发挥一定作用。角质形成细胞(keratinocyte,KC)是人体表皮中的主要细胞,其活化后通过表达一系列粘附分子、趋化因子及细胞因子等,主动参与皮肤的免疫反应,系一种抗原提呈细胞。MF患者皮损的角质形成细胞中某些免疫分子的表达存在着显著不同,如IL-15蛋白的高度表达是皮肤T细胞淋巴瘤的一个特征。 PDCD5 (programmed cell death 5),又名TFAR19 (TF-1 cell apoptosis-related gene 19),是近年新发现的一种凋亡相关基因。该基因进化保守,表达广泛,具有促进细胞凋亡和抑制增殖的效应,是一个潜在的抑癌基因。在多种肿瘤组织中表达均下调,参与肿瘤的发病。但其在皮肤淋巴瘤中的表达情况少见报道。 p53基因是目前研究的较为成熟的与肿瘤相关性最高的抑癌基因。既往研究发现T细胞淋巴瘤中p53基因缺失的突变率为20%-50%,与预后差有明显相关性。亦有学者发现,P53蛋白在MF患者皮损中异常表达,且与端粒酶活化存在相关性,参与MF的发病。本研究通过检测PDCD5与P53蛋白在不同病期MF皮损表皮角质形成细胞及真皮浸润淋巴细胞中表达变化情况及其相关性,初步探讨PDCD5及角质形成细胞在MF发病及病程进展中的作用。目的：检测程序性细胞凋亡因子5(PDCD5)及P53蛋白在不同期蕈样肉芽肿(MF)患者皮损表皮角质形成细胞及真皮淋巴细胞中的表达,初步探讨PDCD5及角质形成细胞在MF发病过程中的作用。材料和方法： 1.选取2006年～2010年山东大学齐鲁医院皮肤科收集的MF组织蜡块24例,其中红斑期19例,斑块/肿瘤期5例,以上病例均经临床、组织病理及免疫组化确诊为MF。取整形美容科及门诊手术室所得10例正常人皮肤组织作为正常对照；另取病理科保存的乳腺癌组织蜡块3例,作为P53的阳性对照。 2.采用免疫组织化学方法分别检测不同期MF及正常对照组标本表皮角质形成细胞及真皮浸润淋巴细胞中PDCD5及P53蛋白的表达。 3.所有数据均应用SPSS13.0统计软件进行分析。PDCD5与P53蛋白在三组标本表皮角质形成细胞及真皮淋巴细胞中表达的阳性等级采用成组设计多样本比较的秩和检验,两蛋白表达的相关性采用Spearman等级相关检验进行分析,P0.05为差异有统计学意义。结果： 1. PDCD5在各组标本中的表达情况 ①正常组皮肤：表皮角质形成细胞及真皮淋巴细胞中PDCD5呈弥漫性强阳性表达,细胞浆和/或细胞核内着棕褐色或棕黄色颗粒； ②红斑期MF：皮损表皮角质形成细胞及真皮浸润淋巴细胞中PDCD5仍呈弥漫性阳性表达,细胞浆和/或细胞核着色,但着色深度较正常对照组浅,呈浅黄或棕黄色； ③斑块/肿瘤期MF：表皮角质形成细胞及真皮浸润淋巴细胞中,PDCD5的表达从阳性细胞所占百分比到着色深度均明显下降,部分病例甚至表达缺如。 ④红斑期MF组与正常对照组皮肤表皮角质形成细胞及真皮淋巴细胞中PDCD5表达差异无统计学意义(P0.05)； ⑤斑块/肿瘤期MF组皮损表皮角质形成细胞及真皮淋巴细胞中PDCD5表达较正常对照组及红斑期MF组均降低(P均0.05)。 2.P53在各组标本中的表达情况 ①正常组皮肤角质形成细胞及真皮淋巴细胞中P53表达均阴性； ②P53在各期MF皮损中呈细胞核着色,棕黄或棕褐色,主要表达于表皮角质形成细胞中,大部分真皮淋巴细胞中缺如,仅1例肿瘤期MF真皮淋巴细胞表达； ③表皮角质形成细胞中正常对照组皮肤表皮角质形成细胞中P53蛋白表达呈阴性；红斑期MF组皮损表皮角质形成细胞中P53蛋白阳性率为36.84%(7/19),斑块/肿瘤期MF组皮损表皮角质形成细胞中P53蛋白阳性率为100%(5/5),三组间任意两组比较,差异均有统计学意义(P均0.05)； ④真皮淋巴细胞中正常对照组和红斑期MF组皮肤真皮淋巴细胞中P53蛋白表达均呈阴性,斑块/肿瘤期MF组皮损真皮淋巴细胞中P53蛋白表达阳性率为20%(1/5),三组间任意两组比较,差异均无统计学意义(P均0.05)。 3.两蛋白相关性分析各期MF的表皮及真皮中PDCD5与P53蛋白的表达均无相关性(P0.05)。结论： 1.随着MF病期的进展,角质形成细胞中PDCD5蛋白的表达明显下调甚至缺如,P53蛋白在其表皮角质形成细胞中的表达逐渐增高,提示角质形成细胞在MF的发病及病程进展中可能发挥作用,具体作用机制有待进一步研究。 2.随着MF病期的进展,真皮淋巴细胞中PDCD5蛋白表达明显下调甚至缺如,提示PDCD5与MF发病及病程进展密切相关,上调PDCD5的表达可能有助于抑制淋巴细胞的过度增殖,PDCD5有望成为治疗MF的新靶点。 3.不同期MF组织中PDCD5与P53蛋白表达无相关性,推测两者可能通过不同的凋亡通路发挥促凋亡效应,也可能与本研究样本例数少,存在一定局限性有关。
[Abstract]:Background of Study :

Fungoides ( MF ) is a low - grade malignant cutaneous T - cell lymphoma derived from memory - assisted T cells . It can be divided into three stages : erythema , plaque and tumor . The pathogenesis of MF is not completely clear . In recent years , the pathogenesis and function of dendritic cells in MF , the homing mechanism of T lymphocytes , the abnormal expression of apoptosis - related genes , such as p53 , p16 and bcl - 2 , have been studied .

It suggests that the interaction of epidermal cells with lymphocytes may play a role in the pathogenesis of MF . Keratinocyte ( KC ) is the main cell in the epidermis of human body . After activation , the expression of certain immune molecules in the keratinocytes of MF patients is significantly different , such as the expression of IL - 15 protein is a feature of cutaneous T - cell lymphoma .

PDCD5 ( programmed cell death 5 ) , also known as TFAR19 ( TF - 1 cell apoptosis - related gene 19 ) , is a newly discovered apoptosis - related gene in recent years . The gene has been evolutionarily conserved , has extensive expression , and has the effect of promoting cell apoptosis and inhibiting proliferation . It is a potential tumor suppressor gene . The expression of PDCD5 in various tumor tissues is downregulated and involved in the pathogenesis of tumor .

The p53 gene is the most mature suppressor gene which has the highest correlation with tumor . Previous research has found that the mutation rate of p53 gene deletion in T cell lymphoma is 20 % -50 % , which is related to poor prognosis . It is also found that P53 protein is abnormal in the skin lesion of MF patients , and it is related to telomerase activation and participates in the pathogenesis of MF .

In this study , the expression of PDCD5 and P53 protein in the epidermal keratinocytes and dermal infiltrating lymphocytes of MF skin lesions of different disease stages were examined . The roles of PDCD5 and keratinocytes in the pathogenesis of MF and progression of the disease were preliminarily discussed .

Purpose :

The expression of apoptosis factor 5 ( PDCD5 ) and P53 protein in epidermal keratinocytes and dermal lymphocytes in patients with MF were examined . The roles of PDCD5 and keratinocytes in the pathogenesis of MF were discussed .

Materials and methods :

1 . There were 24 cases of MF tissue wax collected from Dermatology Department of Shandong University of Shandong University from 2006 to 2010 , including 19 cases of erythema stage , 5 cases of plaque / tumor stage . All the above cases were diagnosed as MF by clinical , histopathological and immunohistochemical method .
3 cases of breast cancer tissue wax block preserved in the department of pathology were taken as positive control of P53 .

2 . The expression of PDCD5 and P53 protein in epidermal keratinocytes and dermis - infiltrating lymphocytes was detected by immunohistochemistry .

3 . All the data were analyzed by SPSS 13.0 . The expression of PDCD5 and P53 protein in epidermal keratinocytes and dermal lymphocytes of three groups were analyzed by using the rank sum test of different groups .

Results :

1 . Expression of PDCD5 in each group

( 1 ) The normal group of skin : epidermal keratinocytes and PDCD5 in the dermal lymphocytes showed diffuse strong positive expression , and the cytoplasm and / or the nucleus were brown or brown - yellow granules ;

( 2 ) The expression of PDCD5 in the epidermis keratinocytes and the dermal infiltrating lymphocytes was diffuse positive , and the cytoplasm and / or nucleus were colored , but the staining depth was lower than that of the normal control group , which was pale yellow or brownish yellow ;

The expression of PDCD5 decreased from the percentage of positive cells to the staining depth , and some cases even expressed absent .

( 4 ) There was no significant difference in the expression of PDCD5 in the keratinocytes and dermal lymphocytes ( P0.05 ) .

( 5 ) The expression of PDCD5 in the epidermis keratinocytes and dermal lymphocytes of the plaque / tumor stage MF group was lower than that in the normal control group and the red spot MF group ( P < 0.05 ) .

2 . Expression of P53 in each group of specimens

The expression of P53 was negative in normal skin keratinocytes and dermal lymphocytes .

( 2 ) P53 was nucleus - colored , brown - yellow or brown - brown in MF lesions of each stage , mainly expressed in epidermal keratinocytes , and most of the dermal lymphocytes were absent , only 1 case was expressed in MF dermal lymphocytes ;

( 3 ) epidermal keratinocytes

The expression of P53 protein was negative in the keratinocytes of normal control group .
The positive rate of P53 protein was 36.84 % ( 7 / 19 ) , the positive rate of P53 protein was 100 % ( 5 / 5 ) in plaque / tumor stage MF group , and the difference was statistically significant ( P < 0.05 ) .

( 4 ) In dermal lymphocytes

The expression of P53 protein was negative in normal control group and red spot MF group , and the positive rate of P53 protein was 20 % ( 1 / 5 ) in plaque / tumor stage MF group , and no significant difference was found between the three groups ( P < 0.05 ) .

3 . Two - protein correlation analysis

There was no correlation between the expression of PDCD5 and P53 protein in epidermis and dermis of MF in each stage ( P0.05 ) .

Conclusion :

1 . With the development of MF disease stage , the expression of PDCD5 protein in keratinocytes was down - regulated and even absent , the expression of P53 protein in keratinocytes was gradually increased , suggesting that keratinocytes might play a role in the pathogenesis of MF and progress of course , and the specific mechanism of action was to be further studied .

2 . With the progression of MF disease stage , the expression of PDCD5 protein in the dermal lymphocytes was down - regulated or even absent , suggesting that PDCD5 is closely related to the pathogenesis of MF and the progression of the disease . The up - regulation of PDCD5 may help to inhibit the excessive proliferation of lymphocytes , and PDCD5 is expected to be a new target for MF .

3 . There was no correlation between PDCD5 and P53 protein expression in MF tissues in the same period . It was speculated that both might play pro - apoptotic effect through different apoptotic pathways , and might be related to the limited number of samples in this study .
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.5

【参考文献】