發(fā)布時(shí)間：2018-06-15 19:37

  本文選題：程序性細(xì)胞凋亡因子5 + 蕈樣肉芽腫��； 參考：《山東大學(xué)》2011年碩士論文

【摘要】：研究背景： 蕈樣肉芽腫,又名蕈樣霉菌病(mycosis fungoides, MF)是起源于記憶性輔助性T細(xì)胞的低度惡性皮膚T細(xì)胞淋巴瘤,臨床可分為紅斑、斑塊、腫瘤三期,病程呈慢性進(jìn)行性,晚期可以侵犯全身各個(gè)臟器。MF的發(fā)病機(jī)制目前尚未完全明確,近年來(lái)對(duì)MF中樹(shù)突狀細(xì)胞的分布及功能、皮膚T淋巴細(xì)胞的歸巢機(jī)制、T淋巴細(xì)胞凋亡障礙及凋亡相關(guān)基因如p53、p16、bcl-2等異常表達(dá)的研究,為MF的發(fā)病機(jī)制及治療提供了一些新的思路。 親表皮現(xiàn)象是MF組織病理學(xué)的特征之一,這提示表皮細(xì)胞與淋巴細(xì)胞的相互作用可能在MF的發(fā)病中發(fā)揮一定作用。角質(zhì)形成細(xì)胞(keratinocyte,KC)是人體表皮中的主要細(xì)胞,其活化后通過(guò)表達(dá)一系列粘附分子、趨化因子及細(xì)胞因子等,主動(dòng)參與皮膚的免疫反應(yīng),系一種抗原提呈細(xì)胞。MF患者皮損的角質(zhì)形成細(xì)胞中某些免疫分子的表達(dá)存在著顯著不同,如IL-15蛋白的高度表達(dá)是皮膚T細(xì)胞淋巴瘤的一個(gè)特征。 PDCD5 (programmed cell death 5),又名TFAR19 (TF-1 cell apoptosis-related gene 19),是近年新發(fā)現(xiàn)的一種凋亡相關(guān)基因。該基因進(jìn)化保守,表達(dá)廣泛,具有促進(jìn)細(xì)胞凋亡和抑制增殖的效應(yīng),是一個(gè)潛在的抑癌基因。在多種腫瘤組織中表達(dá)均下調(diào),參與腫瘤的發(fā)病。但其在皮膚淋巴瘤中的表達(dá)情況少見(jiàn)報(bào)道。 p53基因是目前研究的較為成熟的與腫瘤相關(guān)性最高的抑癌基因。既往研究發(fā)現(xiàn)T細(xì)胞淋巴瘤中p53基因缺失的突變率為20%-50%,與預(yù)后差有明顯相關(guān)性。亦有學(xué)者發(fā)現(xiàn),P53蛋白在MF患者皮損中異常表達(dá),且與端粒酶活化存在相關(guān)性,參與MF的發(fā)病。 本研究通過(guò)檢測(cè)PDCD5與P53蛋白在不同病期MF皮損表皮角質(zhì)形成細(xì)胞及真皮浸潤(rùn)淋巴細(xì)胞中表達(dá)變化情況及其相關(guān)性,初步探討PDCD5及角質(zhì)形成細(xì)胞在MF發(fā)病及病程進(jìn)展中的作用。 目的： 檢測(cè)程序性細(xì)胞凋亡因子5(PDCD5)及P53蛋白在不同期蕈樣肉芽腫(MF)患者皮損表皮角質(zhì)形成細(xì)胞及真皮淋巴細(xì)胞中的表達(dá),初步探討PDCD5及角質(zhì)形成細(xì)胞在MF發(fā)病過(guò)程中的作用。 材料和方法： 1.選取2006年～2010年山東大學(xué)齊魯醫(yī)院皮膚科收集的MF組織蠟塊24例,其中紅斑期19例,斑塊/腫瘤期5例,以上病例均經(jīng)臨床、組織病理及免疫組化確診為MF。取整形美容科及門(mén)診手術(shù)室所得10例正常人皮膚組織作為正常對(duì)照；另取病理科保存的乳腺癌組織蠟塊3例,作為P53的陽(yáng)性對(duì)照。 2.采用免疫組織化學(xué)方法分別檢測(cè)不同期MF及正常對(duì)照組標(biāo)本表皮角質(zhì)形成細(xì)胞及真皮浸潤(rùn)淋巴細(xì)胞中PDCD5及P53蛋白的表達(dá)。 3.所有數(shù)據(jù)均應(yīng)用SPSS13.0統(tǒng)計(jì)軟件進(jìn)行分析。PDCD5與P53蛋白在三組標(biāo)本表皮角質(zhì)形成細(xì)胞及真皮淋巴細(xì)胞中表達(dá)的陽(yáng)性等級(jí)采用成組設(shè)計(jì)多樣本比較的秩和檢驗(yàn),兩蛋白表達(dá)的相關(guān)性采用Spearman等級(jí)相關(guān)檢驗(yàn)進(jìn)行分析,P0.05為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果： 1. PDCD5在各組標(biāo)本中的表達(dá)情況 ①正常組皮膚：表皮角質(zhì)形成細(xì)胞及真皮淋巴細(xì)胞中PDCD5呈彌漫性強(qiáng)陽(yáng)性表達(dá),細(xì)胞漿和/或細(xì)胞核內(nèi)著棕褐色或棕黃色顆粒； ②紅斑期MF：皮損表皮角質(zhì)形成細(xì)胞及真皮浸潤(rùn)淋巴細(xì)胞中PDCD5仍呈彌漫性陽(yáng)性表達(dá),細(xì)胞漿和/或細(xì)胞核著色,但著色深度較正常對(duì)照組淺,呈淺黃或棕黃色； ③斑塊/腫瘤期MF：表皮角質(zhì)形成細(xì)胞及真皮浸潤(rùn)淋巴細(xì)胞中,PDCD5的表達(dá)從陽(yáng)性細(xì)胞所占百分比到著色深度均明顯下降,部分病例甚至表達(dá)缺如。 ④紅斑期MF組與正常對(duì)照組皮膚表皮角質(zhì)形成細(xì)胞及真皮淋巴細(xì)胞中PDCD5表達(dá)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)； ⑤斑塊/腫瘤期MF組皮損表皮角質(zhì)形成細(xì)胞及真皮淋巴細(xì)胞中PDCD5表達(dá)較正常對(duì)照組及紅斑期MF組均降低(P均0.05)。 2.P53在各組標(biāo)本中的表達(dá)情況 ①正常組皮膚角質(zhì)形成細(xì)胞及真皮淋巴細(xì)胞中P53表達(dá)均陰性； ②P53在各期MF皮損中呈細(xì)胞核著色,棕黃或棕褐色,主要表達(dá)于表皮角質(zhì)形成細(xì)胞中,大部分真皮淋巴細(xì)胞中缺如,僅1例腫瘤期MF真皮淋巴細(xì)胞表達(dá)； ③表皮角質(zhì)形成細(xì)胞中 正常對(duì)照組皮膚表皮角質(zhì)形成細(xì)胞中P53蛋白表達(dá)呈陰性；紅斑期MF組皮損表皮角質(zhì)形成細(xì)胞中P53蛋白陽(yáng)性率為36.84%(7/19),斑塊/腫瘤期MF組皮損表皮角質(zhì)形成細(xì)胞中P53蛋白陽(yáng)性率為100%(5/5),三組間任意兩組比較,差異均有統(tǒng)計(jì)學(xué)意義(P均0.05)； ④真皮淋巴細(xì)胞中 正常對(duì)照組和紅斑期MF組皮膚真皮淋巴細(xì)胞中P53蛋白表達(dá)均呈陰性,斑塊/腫瘤期MF組皮損真皮淋巴細(xì)胞中P53蛋白表達(dá)陽(yáng)性率為20%(1/5),三組間任意兩組比較,差異均無(wú)統(tǒng)計(jì)學(xué)意義(P均0.05)。 3.兩蛋白相關(guān)性分析 各期MF的表皮及真皮中PDCD5與P53蛋白的表達(dá)均無(wú)相關(guān)性(P0.05)。 結(jié)論： 1.隨著MF病期的進(jìn)展,角質(zhì)形成細(xì)胞中PDCD5蛋白的表達(dá)明顯下調(diào)甚至缺如,P53蛋白在其表皮角質(zhì)形成細(xì)胞中的表達(dá)逐漸增高,提示角質(zhì)形成細(xì)胞在MF的發(fā)病及病程進(jìn)展中可能發(fā)揮作用,具體作用機(jī)制有待進(jìn)一步研究。 2.隨著MF病期的進(jìn)展,真皮淋巴細(xì)胞中PDCD5蛋白表達(dá)明顯下調(diào)甚至缺如,提示PDCD5與MF發(fā)病及病程進(jìn)展密切相關(guān),上調(diào)PDCD5的表達(dá)可能有助于抑制淋巴細(xì)胞的過(guò)度增殖,PDCD5有望成為治療MF的新靶點(diǎn)。 3.不同期MF組織中PDCD5與P53蛋白表達(dá)無(wú)相關(guān)性,推測(cè)兩者可能通過(guò)不同的凋亡通路發(fā)揮促凋亡效應(yīng),也可能與本研究樣本例數(shù)少,存在一定局限性有關(guān)。
[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 .
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R739.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李惠平,曹志敏,邵玉霞,孫武,賈廷珍;重組人TFAR19蛋白對(duì)羥基喜樹(shù)堿誘導(dǎo)人7721肝癌細(xì)胞凋亡的增敏作用[J];北京醫(yī)科大學(xué)學(xué)報(bào);2000年05期

2 劉朝暉,張岱,李克敏,廖秦平;PDCD5蛋白在正常宮頸、宮頸上皮內(nèi)瘤樣病變和宮頸癌中的表達(dá)[J];北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2004年04期

3 趙曉一,劉宏偉,魏明潔;PDCD5和p53在口腔白斑和口腔鱗癌中表達(dá)的相關(guān)性[J];北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2005年04期

4 孫春玲,喬杰,胡振興,張婷,陳英玉;多囊卵巢綜合征患者黃素化顆粒細(xì)胞中PDCD5蛋白的表達(dá)[J];北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2005年05期

5 宋泉聲,韓文玲,劉紅濤,馬大龍;小鼠凋亡相關(guān)新基因TFAR19 cDNA的克隆化和序列分析[J];北京醫(yī)科大學(xué)學(xué)報(bào);1999年04期

6 趙麗娜;張秀軍;寧樹(shù)成;賀寶玲;趙杰;;系統(tǒng)性紅斑狼瘡患者外周血單個(gè)核細(xì)胞中PDCD5 mRNA的表達(dá)及意義[J];重慶醫(yī)學(xué);2010年15期

7 熊林;譚萬(wàn)龍;郁兆存;吳元東;黃河;趙國(guó)志;朱文輝;鄭少斌;;TFAR19(PDCD5)蛋白在正常腎、腎透明細(xì)胞癌組織及正常膀胱、膀胱癌組織中的表達(dá)[J];南方醫(yī)科大學(xué)學(xué)報(bào);2006年06期

8 馮靜,崔恒,魏麗惠,馬大龍;TFAR19蛋白在卵巢上皮性癌中的表達(dá)[J];中國(guó)婦產(chǎn)科臨床;2002年03期

9 王靖雪;p53和淋巴系統(tǒng)惡性腫瘤[J];國(guó)外醫(yī)學(xué)(兒科學(xué)分冊(cè));2001年03期

10 杜彥丹;于靖;趙雪飛;林平;劉紫君;于曉光;;PDCD5蛋白聯(lián)合米非司酮對(duì)前列腺癌細(xì)胞增殖及相關(guān)基因表達(dá)的影響[J];腫瘤基礎(chǔ)與臨床;2009年02期

，

本文編號(hào)：2023310