APOOBEC3G蛋白在尖銳濕疣及幾種皮膚腫瘤中的表達(dá)
發(fā)布時(shí)間：2018-09-04 18:41
【摘要】： 前言 人乳頭瘤病毒(human papilloma vivus, HPV)是一種有種屬和組織特異性的雙鏈閉環(huán)的脫氧核糖核酸(DNA)病毒,可引起肛門周圍、外陰生殖器的良惡性損害。HPV病毒持續(xù)性感染是造成尖銳濕疣(human papilloma vivus, CA)復(fù)發(fā)或?qū)m頸癌發(fā)生的重要原因,而HPV感染后局部細(xì)胞免疫功能低下是HPV持續(xù)性感染的根本原因所在。 載脂蛋白B mRNA編輯酶催化多肽樣蛋白3G(apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 protein G, APOBEC3G)是在人類免疫缺陷病毒的研究過程中發(fā)現(xiàn)的一種人類特有的天然免疫因子,屬于固有免疫家族成員。它不僅能抑制人類免疫缺陷病毒的復(fù)制,還能在瞬時(shí)表達(dá)乙型肝炎病毒的共轉(zhuǎn)染細(xì)胞體系中發(fā)揮顯著抑制病毒復(fù)制的生物學(xué)功能,并對(duì)來自不同宿主的其它逆轉(zhuǎn)錄病毒成員如鼠白血病病毒、猿免疫缺陷病毒等均具有抑制作用。因此,APOBEC3G具有廣譜的抗病毒活性。但由于APOBEC3G的基因結(jié)構(gòu)及蛋白功能與胞嘧啶脫氨酶家族其它成員非常相似,其基因過度表達(dá)可能會(huì)導(dǎo)致細(xì)胞內(nèi)基因組不穩(wěn)定性增加,從而參與惡性腫瘤的發(fā)生發(fā)展過程。 本研究觀察AOPBEC3G蛋白在人體CA、某些皮膚惡性腫瘤等組織中的表達(dá)情況,并以銀屑病及正常皮膚組織作為對(duì)照,以初步探討APOBEC3G在HPV感染相關(guān)皮膚病中的作用。 材料和方法 一、研究對(duì)象 1、病史采集完整的CA組織12例,鱗狀細(xì)胞癌組織、基底細(xì)胞癌組織、角化棘皮瘤組織、銀屑病組織各10例,正常皮膚組織5例。 2、各種組織病例性別及年齡具有可比性,無系統(tǒng)性疾病或其它皮膚病。 二、材料 1、APOBEC3G抗體(ab71634, ABCOM, UK)即用型S-P免疫組化試劑盒 PBS緩沖液 2、3-二氨基聯(lián)苯胺(DAB)顯色系統(tǒng) 三、實(shí)驗(yàn)方法 制備6u厚石蠟切片�？乖邏簾嵝迯�(fù)。先后滴加兔抗人多克隆抗EBA一抗,PBS作陰性對(duì)照。DAB顯色,蘇木素復(fù)染。脫水、透明、封片。光鏡下觀察。 四、結(jié)果判定 光鏡下(x400倍),每張切片隨機(jī)選取4個(gè)視野,以細(xì)胞膜或胞漿內(nèi)出現(xiàn)棕黃色或棕褐色顆粒為陽(yáng)性。采用半定量積分法,對(duì)細(xì)胞中陽(yáng)性細(xì)胞的染色強(qiáng)度及陽(yáng)性細(xì)胞數(shù)進(jìn)行人工判定；顯微圖像分析系統(tǒng)采集圖像,分析陽(yáng)性表達(dá)積分光密度。取平均值,以積分光密度／高倍視野為單位計(jì)算,獲得每例標(biāo)本陽(yáng)性染色的光密度值及陽(yáng)性百分比。 五、統(tǒng)計(jì)學(xué)分析 采用SPSS16.0軟件包進(jìn)行數(shù)據(jù)處理。分類資料采用X2檢驗(yàn),計(jì)量資料采用LSD-t檢驗(yàn)。P0.05認(rèn)為有統(tǒng)計(jì)學(xué)差異。 結(jié)果 1、12例CA組織中,9例呈高表達(dá),1例弱表達(dá),陽(yáng)性率為91.67%；10例SCC及10例BCC組織中,各有4例呈高表達(dá),各有2例弱表達(dá),陽(yáng)性率均為60%；10例KA組織中,5例呈高表達(dá),2例弱表達(dá),陽(yáng)性率均為70%。10例銀屑病中,有1例陽(yáng)性表達(dá)。正常皮膚無表達(dá)。 2、CA、SCC、BCC、KA陽(yáng)性表達(dá)分布情況均與對(duì)照組織(銀屑病及正常皮膚)具有統(tǒng)計(jì)學(xué)差異。CA及各種惡性皮膚腫瘤間陽(yáng)性表達(dá)無統(tǒng)計(jì)學(xué)差異,銀屑病與正常皮膚無統(tǒng)計(jì)學(xué)差異。 3、CA組織光密度值及陽(yáng)性百分比最高,分別為17.48及33.51%；銀屑病及正常皮膚組織最低,分別約為8.51%及7.82%；SCC、BCC、KA界于中間。 4、CA、SCC、BCC、KA的光密度值及陽(yáng)性百分比均與對(duì)照組織(銀屑病及正常皮膚)具有統(tǒng)計(jì)學(xué)差異。CA與三種皮膚惡性腫瘤之間有統(tǒng)計(jì)學(xué)差異。三種皮膚腫瘤之間無統(tǒng)計(jì)學(xué)差異,銀屑病與正常皮膚組織無統(tǒng)計(jì)學(xué)差異。 結(jié)論 1) APOBEC3G蛋白在大部分HPV感染的CA組織中,呈高水平表達(dá)。 2) APOBEC3G蛋白在SCC、BCC、KA組織中,呈中等水平表達(dá)。 3) APOBEC3G蛋白在銀屑病及正常皮膚組織中,呈低水平表達(dá)。
[Abstract]:Preface
Human papillomavirus (HPV) is a species-and tissue-specific double-stranded closed-loop deoxyribonucleic acid (DNA) virus that can cause benign and malignant damage to genitals around the anus and vulva. However, local cellular immune function after HPV infection is the root cause of persistent infection of HPV.
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 protein G (APOBEC3G) is a kind of human specific natural immune factor found in the research of human immunodeficiency virus. It belongs to the innate immune family. It can not only inhibit human immunodeficiency diseases. The replication of the virus can also play a significant role in inhibiting the replication of hepatitis B virus in the co-transfected cell system with transient expression of hepatitis B virus, and inhibit other retroviral members from different hosts, such as mouse leukemia virus, simian immunodeficiency virus, etc. Therefore, APOBEC3G has broad-spectrum antiviral activity. Because APOBEC3G gene structure and protein function are very similar to other members of the cytosine deaminase family, overexpression of APOBEC3G gene may lead to increased intracellular genomic instability, thus participating in the development of malignant tumors.
In this study, we observed the expression of AOPBEC3G protein in human CA, some skin malignant tumors and other tissues, and compared with psoriasis and normal skin tissues to explore the role of APOBEC3G in HPV infection-related skin diseases.
Materials and methods
First, the object of study.
1. A total of 12 cases of CA, squamous cell carcinoma, basal cell carcinoma, keratoacanthoma, psoriasis and 5 normal skin tissues were collected.
2, the sex and age of various tissues are comparable, without systemic diseases or other skin diseases.
Two, material
1, APOBEC3G antibody (ab71634, ABCOM, UK) instant S-P immunohistochemistry kit.
PBS buffer solution
2,3- two amino benzidine (DAB) color rendering system
Three, the experimental method.
Six u thick paraffin sections were prepared. Antigen was repaired by hyperbaric heating. Rabbit anti-Human Polyclonal anti-EBA antibody was dripped successively. PBS was used as negative control. DAB was developed, hematoxylin was re-stained, dehydrated, transparent and sealed. The results were observed under light microscope.
Four, result judgement
Under the light microscope (x 400 times), each slice was randomly selected from four visual fields, and the brown-yellow or brown granules in the cell membrane or cytoplasm were taken as positive. The intensity of staining and the number of positive cells in the cells were determined by semi-quantitative integration method. The average value of optical density and positive percentage of positive staining were obtained by calculating integral optical density/high power visual field.
Five, statistical analysis.
SPSS16.0 software package was used for data processing. X2 test was used for classification data and LSD-t test was used for measurement data. P 0.05 showed that there was a statistical difference.
Result
In 1,12 cases of CA, 9 cases were overexpressed, 1 case was weakly expressed, the positive rate was 91.67%; in 10 cases of SCC and 10 cases of BCC, 4 cases were overexpressed, 2 cases were weakly expressed, the positive rate was 60%; in 10 cases of KA, 5 cases were overexpressed, 2 cases were weakly expressed, the positive rate was 70%.
2. The distribution of positive expression of CA, SCC, BCC and KA was significantly different from that of the control group (psoriasis and normal skin). The positive expression of CA and various malignant skin tumors was not significantly different, and there was no significant difference between psoriasis and normal skin.
3. The highest optical density and positive percentage of CA were 17.48% and 33.51%, respectively; the lowest were psoriasis and normal skin tissues, about 8.51% and 7.82%, respectively; SCC, BCC, KA were in the middle.
4. The optical density values and positive percentages of CA, SCC, BCC and KA were significantly different from those of the control group (psoriasis and normal skin). There were significant differences between CA and three kinds of skin malignant tumors. There was no significant difference among the three kinds of skin tumors, and no significant difference between psoriasis and normal skin tissue.
conclusion
1) APOBEC3G protein is highly expressed in most HPV infected CA tissues.
2) APOBEC3G protein was moderately expressed in SCC, BCC and KA tissues.
3) APOBEC3G protein was expressed at low level in psoriasis and normal skin tissues.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R752.53
[Abstract]:Preface
Human papillomavirus (HPV) is a species-and tissue-specific double-stranded closed-loop deoxyribonucleic acid (DNA) virus that can cause benign and malignant damage to genitals around the anus and vulva. However, local cellular immune function after HPV infection is the root cause of persistent infection of HPV.
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 protein G (APOBEC3G) is a kind of human specific natural immune factor found in the research of human immunodeficiency virus. It belongs to the innate immune family. It can not only inhibit human immunodeficiency diseases. The replication of the virus can also play a significant role in inhibiting the replication of hepatitis B virus in the co-transfected cell system with transient expression of hepatitis B virus, and inhibit other retroviral members from different hosts, such as mouse leukemia virus, simian immunodeficiency virus, etc. Therefore, APOBEC3G has broad-spectrum antiviral activity. Because APOBEC3G gene structure and protein function are very similar to other members of the cytosine deaminase family, overexpression of APOBEC3G gene may lead to increased intracellular genomic instability, thus participating in the development of malignant tumors.
In this study, we observed the expression of AOPBEC3G protein in human CA, some skin malignant tumors and other tissues, and compared with psoriasis and normal skin tissues to explore the role of APOBEC3G in HPV infection-related skin diseases.
Materials and methods
First, the object of study.
1. A total of 12 cases of CA, squamous cell carcinoma, basal cell carcinoma, keratoacanthoma, psoriasis and 5 normal skin tissues were collected.
2, the sex and age of various tissues are comparable, without systemic diseases or other skin diseases.
Two, material
1, APOBEC3G antibody (ab71634, ABCOM, UK) instant S-P immunohistochemistry kit.
PBS buffer solution
2,3- two amino benzidine (DAB) color rendering system
Three, the experimental method.
Six u thick paraffin sections were prepared. Antigen was repaired by hyperbaric heating. Rabbit anti-Human Polyclonal anti-EBA antibody was dripped successively. PBS was used as negative control. DAB was developed, hematoxylin was re-stained, dehydrated, transparent and sealed. The results were observed under light microscope.
Four, result judgement
Under the light microscope (x 400 times), each slice was randomly selected from four visual fields, and the brown-yellow or brown granules in the cell membrane or cytoplasm were taken as positive. The intensity of staining and the number of positive cells in the cells were determined by semi-quantitative integration method. The average value of optical density and positive percentage of positive staining were obtained by calculating integral optical density/high power visual field.
Five, statistical analysis.
SPSS16.0 software package was used for data processing. X2 test was used for classification data and LSD-t test was used for measurement data. P 0.05 showed that there was a statistical difference.
Result
In 1,12 cases of CA, 9 cases were overexpressed, 1 case was weakly expressed, the positive rate was 91.67%; in 10 cases of SCC and 10 cases of BCC, 4 cases were overexpressed, 2 cases were weakly expressed, the positive rate was 60%; in 10 cases of KA, 5 cases were overexpressed, 2 cases were weakly expressed, the positive rate was 70%.
2. The distribution of positive expression of CA, SCC, BCC and KA was significantly different from that of the control group (psoriasis and normal skin). The positive expression of CA and various malignant skin tumors was not significantly different, and there was no significant difference between psoriasis and normal skin.
3. The highest optical density and positive percentage of CA were 17.48% and 33.51%, respectively; the lowest were psoriasis and normal skin tissues, about 8.51% and 7.82%, respectively; SCC, BCC, KA were in the middle.
4. The optical density values and positive percentages of CA, SCC, BCC and KA were significantly different from those of the control group (psoriasis and normal skin). There were significant differences between CA and three kinds of skin malignant tumors. There was no significant difference among the three kinds of skin tumors, and no significant difference between psoriasis and normal skin tissue.
conclusion
1) APOBEC3G protein is highly expressed in most HPV infected CA tissues.
2) APOBEC3G protein was moderately expressed in SCC, BCC and KA tissues.
3) APOBEC3G protein was expressed at low level in psoriasis and normal skin tissues.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R752.53
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