APOOBEC3G蛋白在尖锐湿疣及几种皮肤肿瘤中的表达
发布时间：2018-09-04 18:41
【摘要】： 前言 人乳头瘤病毒(human papilloma vivus, HPV)是一种有种属和组织特异性的双链闭环的脱氧核糖核酸(DNA)病毒,可引起肛门周围、外阴生殖器的良恶性损害。HPV病毒持续性感染是造成尖锐湿疣(human papilloma vivus, CA)复发或宫颈癌发生的重要原因,而HPV感染后局部细胞免疫功能低下是HPV持续性感染的根本原因所在。 载脂蛋白B mRNA编辑酶催化多肽样蛋白3G(apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 protein G, APOBEC3G)是在人类免疫缺陷病毒的研究过程中发现的一种人类特有的天然免疫因子,属于固有免疫家族成员。它不仅能抑制人类免疫缺陷病毒的复制,还能在瞬时表达乙型肝炎病毒的共转染细胞体系中发挥显著抑制病毒复制的生物学功能,并对来自不同宿主的其它逆转录病毒成员如鼠白血病病毒、猿免疫缺陷病毒等均具有抑制作用。因此,APOBEC3G具有广谱的抗病毒活性。但由于APOBEC3G的基因结构及蛋白功能与胞嘧啶脱氨酶家族其它成员非常相似,其基因过度表达可能会导致细胞内基因组不稳定性增加,从而参与恶性肿瘤的发生发展过程。 本研究观察AOPBEC3G蛋白在人体CA、某些皮肤恶性肿瘤等组织中的表达情况,并以银屑病及正常皮肤组织作为对照,以初步探讨APOBEC3G在HPV感染相关皮肤病中的作用。 材料和方法 一、研究对象 1、病史采集完整的CA组织12例,鳞状细胞癌组织、基底细胞癌组织、角化棘皮瘤组织、银屑病组织各10例,正常皮肤组织5例。 2、各种组织病例性别及年龄具有可比性,无系统性疾病或其它皮肤病。 二、材料 1、APOBEC3G抗体(ab71634, ABCOM, UK)即用型S-P免疫组化试剂盒 PBS缓冲液 2、3-二氨基联苯胺(DAB)显色系统 三、实验方法 制备6u厚石蜡切片。抗原高压热修复。先后滴加兔抗人多克隆抗EBA一抗,PBS作阴性对照。DAB显色,苏木素复染。脱水、透明、封片。光镜下观察。 四、结果判定 光镜下(x400倍),每张切片随机选取4个视野,以细胞膜或胞浆内出现棕黄色或棕褐色颗粒为阳性。采用半定量积分法,对细胞中阳性细胞的染色强度及阳性细胞数进行人工判定；显微图像分析系统采集图像,分析阳性表达积分光密度。取平均值,以积分光密度／高倍视野为单位计算,获得每例标本阳性染色的光密度值及阳性百分比。 五、统计学分析 采用SPSS16.0软件包进行数据处理。分类资料采用X2检验,计量资料采用LSD-t检验。P0.05认为有统计学差异。 结果 1、12例CA组织中,9例呈高表达,1例弱表达,阳性率为91.67%；10例SCC及10例BCC组织中,各有4例呈高表达,各有2例弱表达,阳性率均为60%；10例KA组织中,5例呈高表达,2例弱表达,阳性率均为70%。10例银屑病中,有1例阳性表达。正常皮肤无表达。 2、CA、SCC、BCC、KA阳性表达分布情况均与对照组织(银屑病及正常皮肤)具有统计学差异。CA及各种恶性皮肤肿瘤间阳性表达无统计学差异,银屑病与正常皮肤无统计学差异。 3、CA组织光密度值及阳性百分比最高,分别为17.48及33.51%；银屑病及正常皮肤组织最低,分别约为8.51%及7.82%；SCC、BCC、KA界于中间。 4、CA、SCC、BCC、KA的光密度值及阳性百分比均与对照组织(银屑病及正常皮肤)具有统计学差异。CA与三种皮肤恶性肿瘤之间有统计学差异。三种皮肤肿瘤之间无统计学差异,银屑病与正常皮肤组织无统计学差异。 结论 1) APOBEC3G蛋白在大部分HPV感染的CA组织中,呈高水平表达。 2) APOBEC3G蛋白在SCC、BCC、KA组织中,呈中等水平表达。 3) APOBEC3G蛋白在银屑病及正常皮肤组织中,呈低水平表达。
[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.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：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.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R752.53
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