本文選題：免疫球蛋白M + 鼻咽部腫瘤　； 參考：《瀘州醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的：隨著各種診斷方式的更新及治療方法的進(jìn)步，鼻咽癌目前的5年生存率有了一定提高，但由于鼻咽癌是一類多因素導(dǎo)致的疾病，其發(fā)病機制仍不明確，早期診斷困難，總體治療效果不理想。在前期研究中，我們發(fā)現(xiàn)免疫球蛋白（Immunoglobulin，Ig）M在多種頭頸部腫瘤包括鼻咽癌中存在異位表達(dá)，利用抗人IgM抗體干預(yù)后人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤的生長受到明顯抑制；我們推測IgM對人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤具有生長因子樣作用，但其具體機制尚不明確[1,2]。本實驗是在前期研究的基礎(chǔ)上，獲取人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤組織標(biāo)本，通過檢測移植瘤組織中生存素（Survivin）、增殖細(xì)胞核抗原（proliferating cellnuclear antigen，PCNA）、血管生成素-2（angiopoietin，Ang-2）以及移植瘤內(nèi)微血管密度（microvessel density，MVD）計數(shù)情況，進(jìn)一步探討抗人IgM抗體抑制人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤生長可能的機制，從而為鼻咽癌發(fā)病機制研究以及將抗人IgM抗體應(yīng)用于鼻咽癌的臨床治療提供理論依據(jù)。方法：取人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤組織，用石蠟包埋瘤體組織，輪轉(zhuǎn)式切片機將其切成4μm厚的病理切片，應(yīng)用免疫組化鏈霉親和素-生物素化過氧化酶復(fù)合物染色法（Streptoavidin-biotin-enzymecomplex，SABC）檢測瘤體組織中Survivin、PCNA、Ang-2表達(dá)情況；染色后在Olympus BH2顯微鏡下采集圖像，每個指標(biāo)各5張切片，每張采集5張圖片，運用Image-pro Plus6.0軟件對圖片作免疫組化半定量分析，記錄平均光密度值。應(yīng)用免疫組化鏈霉親和堿性磷酸酶染色法（Streptavidin-Alkaline Phosphatase，SAP）檢測CD31標(biāo)記的移植瘤內(nèi)微血管，并計數(shù)MVD。實驗結(jié)果均以均數(shù)±標(biāo)準(zhǔn)差（x±s）表達(dá)，統(tǒng)計數(shù)據(jù)運用SPSS16.0for windows軟件作統(tǒng)計學(xué)分析，P0.05表示差異有統(tǒng)計學(xué)意義。結(jié)果：（1）Survivin表達(dá)情況：抗人IgM抗體干預(yù)組（實驗組）Survivin表達(dá)的光密度值為（0.153±0.009），IgG對照組Survivin表達(dá)的光密度值為（0.221±0.019），PBS對照組Survivin表達(dá)的光密度值為（0.246±0.021），實驗組Survivin陽性表達(dá)的結(jié)果顯著低于IgG對照組和PBS對照組，差異有統(tǒng)計學(xué)意義（P0.05）。（2）PCNA表達(dá)情況：抗人IgM抗體干預(yù)組表達(dá)的PCNA平均光密度值為（0.084±0.025），IgG對照組表達(dá)的平均光密度值為（0.149±0.016），PBS對照組PCNA表達(dá)的平均光密度為（0.163±0.018），實驗組PCNA陽性表達(dá)的結(jié)果顯著低于IgG對照組和PBS對照組，差異有統(tǒng)計學(xué)意義（P0.05）。（3）Ang-2表達(dá)情況：抗人IgM抗體干預(yù)組Ang-2表達(dá)的平均光密度值為（0.121±0.021），，IgG對照組Ang-2表達(dá)的平均光密度值為（0.188±0.013），PBS對照組的平均光密度為（0.196±0.005），實驗組Ang-2陽性表達(dá)的結(jié)果顯著低于IgG對照組和PBS對照組，差異有統(tǒng)計學(xué)意義（P0.05）。（4）MVD計數(shù)情況：抗人IgM抗體干預(yù)組MVD為（12.54±0.47），顯著低于IgG對照組（16.95±0.21）及PBS對照組（17.17±0.47），差異有統(tǒng)計學(xué)意義（P0.05）。（5）人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤組織中MVD分別與Survivin和Ang-2呈顯著正相關(guān)關(guān)系，相關(guān)系數(shù)分別為0.754，0.696，P值分別為0.001，0.004。結(jié)論：（1）抗人IgM抗體可以顯著抑制人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤組織內(nèi)Survivin的表達(dá)。（2）抗人IgM抗體可以顯著抑制人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤內(nèi)PCNA的表達(dá)。（3）抗人IgM抗體可以顯著抑制人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤內(nèi)Ang-2的表達(dá)。（4）抗人IgM抗體可以顯著抑制人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤組織MVD，即抑制血管生成，其機制可能與抑制Ang-2、Survivin的表達(dá)有關(guān)。（5）抗人IgM抗體抑制人鼻咽癌HNE-1細(xì)胞裸鼠移植瘤生長的機制可能與其促進(jìn)凋亡、抑制細(xì)胞增殖和血管生成有關(guān)。
[Abstract]:Objective: with the update of various diagnostic methods and the progress of treatment methods, the 5 year survival rate of nasopharyngeal carcinoma has been improved. However, because nasopharyngeal carcinoma is a disease caused by a class of multiple factors, its pathogenesis is still not clear, early diagnosis is difficult, and the overall treatment effect is not ideal. In the early study, we found immunoglobulin (Immunogl Obulin, Ig) M has heterotopic expression in a variety of head and neck tumors including nasopharyngeal carcinoma, and the growth of nude mice transplanted tumor of human nasopharyngeal carcinoma HNE-1 cells after intervention with anti human IgM antibody is obviously inhibited. We speculate that IgM has a growth factor like effect on human nasopharyngeal carcinoma HNE-1 cell xenografts in nude mice, but the specific mechanism is not yet clear in [1,2]. experiment. On the basis of previous studies, the tissue specimens of human nasopharyngeal carcinoma HNE-1 cells transplanted in nude mice were obtained by detecting survivin (Survivin), proliferating cell nuclear antigen (proliferating Cellnuclear antigen, PCNA), angiopoietin -2 (angiopoietin, Ang-2), and microvascular density (microvessel density, MVD) in the transplanted tumor tissue. The possible mechanism of anti human IgM antibody inhibiting the growth of human nasopharyngeal carcinoma HNE-1 cell xenografts in nude mice was further investigated, thus providing a theoretical basis for the study of the pathogenesis of nasopharyngeal carcinoma and the application of anti human IgM antibody in the clinical treatment of nasopharyngeal carcinoma. The weave and rotating slicer cut it into 4 m thick pathological sections, and the expression of Survivin, PCNA, Ang-2 in the tumor tissues was detected by Streptoavidin-biotin-enzymecomplex (SABC) with immunohistochemical streptomycin biotin peroxidase complex (SABC). The images were collected under the Olympus BH2 microscope after staining, and each index was 5 slices each. Each piece, 5 pictures were collected each, and the average light density was recorded by Image-pro Plus6.0 software, and the average light density was recorded. The microvessels in the transplanted tumor with CD31 markers were detected by immuno histochemical Streptomyces affinity alkaline phosphatase staining (Streptavidin-Alkaline Phosphatase, SAP), and the results of MVD. test were all with a mean of average number. X + s expression, statistical data using SPSS16.0for windows software for statistical analysis, P0.05 indicated that the difference was statistically significant. Results: (1) Survivin expression: the light density value of Survivin expression in the anti human IgM antibody group (experimental group) was (0.153 + 0.009), and the optical density value of Survivin expression in IgG control group was (0.221 + 0.019), PBS pair The intensity of light density expressed in group Survivin was (0.246 + 0.021). The positive expression of Survivin in the experimental group was significantly lower than that of the IgG control group and the PBS control group. The difference was statistically significant (P0.05). (2) the expression of PCNA: the mean PCNA of the anti human IgM antibody group was (0.084 + 0.025), and the average optical density of the IgG control group The average optical density of PCNA expression in the PBS control group was (0.163 + 0.018). The results of PCNA positive expression in the experimental group were significantly lower than those in the IgG control group and the PBS control group. The difference was statistically significant (P0.05). (3) the expression of Ang-2: the average optical density of the Ang-2 expression in the anti human IgM antibody group was (0.121 + 0.021), and the Ang-2 table of the IgG control group The average light density of the PBS group was (0.188 + 0.013), the average light density of the control group was (0.196 + 0.005). The results of Ang-2 positive expression in the experimental group were significantly lower than that of the IgG control group and the PBS control group. (4) the MVD count situation: the MVD in the anti human IgM antibody group was (12.54 + 0.47), significantly lower than that of the IgG control group (16.95 + 0.21). And PBS control group (17.17 + 0.47), the difference was statistically significant (P0.05). (5) there was a significant positive correlation between MVD and Survivin and Ang-2 in human nasopharyngeal carcinoma HNE-1 cells transplanted tumor tissue in nude mice. The correlation coefficient was 0.754,0.696, P value was respectively 0.001,0.004. conclusion: (1) anti human IgM antibody could significantly inhibit the migration of nude mice in nasopharyngeal carcinoma HNE-1 cells. The expression of Survivin in the tumor tissue. (2) anti human IgM antibody can significantly inhibit the expression of PCNA in human nasopharyngeal carcinoma HNE-1 cells transplanted tumor in nude mice. (3) anti human IgM antibody can significantly inhibit the expression of Ang-2 in human nasopharyngeal carcinoma HNE-1 cells transplanted tumor in nude mice. (4) anti human IgM antibody can significantly inhibit the MVD of human nasopharyngeal carcinoma HNE-1 cell nude mice xenografts MVD The mechanism of inhibiting angiogenesis may be related to the inhibition of the expression of Ang-2 and Survivin. (5) the mechanism of anti human IgM antibody inhibiting the growth of human nasopharyngeal carcinoma HNE-1 cell xenografts may be related to the promotion of apoptosis, inhibition of cell proliferation and angiogenesis.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R739.63

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李超,馮紅超,陳建超,宋宇峰;口腔鱗癌組織中Angiopoietin-1、Angiopoietin-2的表達(dá)及臨床意義[J];癌癥;2005年11期

2 王金萬,孫燕,劉永煜,于起濤,張沂平,李凱,朱允中,周清華,侯梅,管忠震,李維廉,莊武,王東林,梁后杰,秦鳳展,盧輝山,劉曉晴,孫紅,張燕軍,王杰軍,羅素霞,楊瑞合,涂遠(yuǎn)榮,王秀問,宋恕平,周靜敏,游麗芬,王競,姚晨;重組人血管內(nèi)皮抑素聯(lián)合NP方案治療晚期NSCLC隨機、雙盲、對照、多中心Ⅲ期臨床研究[J];中國肺癌雜志;2005年04期

3 申洪;免疫組織化學(xué)染色定量方法研究（Ⅲ）[J];中國組織化學(xué)與細(xì)胞化學(xué)雜志;1995年01期

4 劉穎新;朱小東;葛蓮英;王安宇;陳龍;陳發(fā)龍;羅元;;PCNA和PTEN的表達(dá)水平在鼻咽癌預(yù)后判斷中的意義[J];臨床腫瘤學(xué)雜志;2006年10期

5 夏冰;楊太成;冼江;王捷;;鼻咽癌患者血清血管生長素水平的臨床意義初探[J];臨床耳鼻咽喉科雜志;2006年14期

6 李浩勇;梁培育;朱孝峰;鄭克立;戴宇平;;免疫球蛋白G在膀胱癌細(xì)胞中的表達(dá)及其抗體誘導(dǎo)腫瘤細(xì)胞凋亡的研究[J];中國病理生理雜志;2007年06期

7 練兵;王繼群;金琳;;靶向PCNA基因的小干擾RNA對鼻咽癌CNE2細(xì)胞周期的影響[J];中國病理生理雜志;2009年08期

8 王萍;陳群;朱美玉;孟慶勇;唐小梅;孫曉暉;劉瓊玲;王芳;;抗人Ig抗體誘導(dǎo)人乳腺癌MCF-7細(xì)胞凋亡的實驗研究[J];中國現(xiàn)代醫(yī)生;2009年11期

9 張帥,肖健云,田勇泉,趙素萍,邱元正,王承龍;凋亡抑制基因Survivin mRNA在鼻咽癌中的表達(dá)及其臨床意義[J];中國耳鼻咽喉顱底外科雜志;2004年01期

10 黃國森,涂青松,衛(wèi)光宇;鼻咽癌的血管生成及其臨床意義[J];中國耳鼻咽喉顱底外科雜志;2004年01期

相關(guān)博士學(xué)位論文 前1條

1 覃綱;免疫球蛋白M與鼻咽癌的實驗研究[D];四川大學(xué);2007年

本文編號：2036663