抗人IgM抗体抑制人鼻咽癌HNE-1细胞裸鼠移植瘤生长的机制研究

发布时间：2018-06-18 19:49

本文选题：免疫球蛋白M + 鼻咽部肿瘤　；参考：《泸州医学院》2012年硕士论文

【摘要】：目的：随着各种诊断方式的更新及治疗方法的进步，鼻咽癌目前的5年生存率有了一定提高，但由于鼻咽癌是一类多因素导致的疾病，其发病机制仍不明确，早期诊断困难，总体治疗效果不理想。在前期研究中，我们发现免疫球蛋白（Immunoglobulin，Ig）M在多种头颈部肿瘤包括鼻咽癌中存在异位表达，利用抗人IgM抗体干预后人鼻咽癌HNE-1细胞裸鼠移植瘤的生长受到明显抑制；我们推测IgM对人鼻咽癌HNE-1细胞裸鼠移植瘤具有生长因子样作用，但其具体机制尚不明确[1,2]。本实验是在前期研究的基础上，获取人鼻咽癌HNE-1细胞裸鼠移植瘤组织标本，通过检测移植瘤组织中生存素（Survivin）、增殖细胞核抗原（proliferating cellnuclear antigen，PCNA）、血管生成素-2（angiopoietin，Ang-2）以及移植瘤内微血管密度（microvessel density，MVD）计数情况，进一步探讨抗人IgM抗体抑制人鼻咽癌HNE-1细胞裸鼠移植瘤生长可能的机制，从而为鼻咽癌发病机制研究以及将抗人IgM抗体应用于鼻咽癌的临床治疗提供理论依据。方法：取人鼻咽癌HNE-1细胞裸鼠移植瘤组织，用石蜡包埋瘤体组织，轮转式切片机将其切成4μm厚的病理切片，应用免疫组化链霉亲和素-生物素化过氧化酶复合物染色法（Streptoavidin-biotin-enzymecomplex，SABC）检测瘤体组织中Survivin、PCNA、Ang-2表达情况；染色后在Olympus BH2显微镜下采集图像，每个指标各5张切片，每张采集5张图片，运用Image-pro Plus6.0软件对图片作免疫组化半定量分析，记录平均光密度值。应用免疫组化链霉亲和碱性磷酸酶染色法（Streptavidin-Alkaline Phosphatase，SAP）检测CD31标记的移植瘤内微血管，并计数MVD。实验结果均以均数±标准差（x±s）表达，统计数据运用SPSS16.0for windows软件作统计学分析，P0.05表示差异有统计学意义。结果：（1）Survivin表达情况：抗人IgM抗体干预组（实验组）Survivin表达的光密度值为（0.153±0.009），IgG对照组Survivin表达的光密度值为（0.221±0.019），PBS对照组Survivin表达的光密度值为（0.246±0.021），实验组Survivin阳性表达的结果显著低于IgG对照组和PBS对照组，差异有统计学意义（P0.05）。（2）PCNA表达情况：抗人IgM抗体干预组表达的PCNA平均光密度值为（0.084±0.025），IgG对照组表达的平均光密度值为（0.149±0.016），PBS对照组PCNA表达的平均光密度为（0.163±0.018），实验组PCNA阳性表达的结果显著低于IgG对照组和PBS对照组，差异有统计学意义（P0.05）。（3）Ang-2表达情况：抗人IgM抗体干预组Ang-2表达的平均光密度值为（0.121±0.021），，IgG对照组Ang-2表达的平均光密度值为（0.188±0.013），PBS对照组的平均光密度为（0.196±0.005），实验组Ang-2阳性表达的结果显著低于IgG对照组和PBS对照组，差异有统计学意义（P0.05）。（4）MVD计数情况：抗人IgM抗体干预组MVD为（12.54±0.47），显著低于IgG对照组（16.95±0.21）及PBS对照组（17.17±0.47），差异有统计学意义（P0.05）。（5）人鼻咽癌HNE-1细胞裸鼠移植瘤组织中MVD分别与Survivin和Ang-2呈显著正相关关系，相关系数分别为0.754，0.696，P值分别为0.001，0.004。结论：（1）抗人IgM抗体可以显著抑制人鼻咽癌HNE-1细胞裸鼠移植瘤组织内Survivin的表达。（2）抗人IgM抗体可以显著抑制人鼻咽癌HNE-1细胞裸鼠移植瘤内PCNA的表达。（3）抗人IgM抗体可以显著抑制人鼻咽癌HNE-1细胞裸鼠移植瘤内Ang-2的表达。（4）抗人IgM抗体可以显著抑制人鼻咽癌HNE-1细胞裸鼠移植瘤组织MVD，即抑制血管生成，其机制可能与抑制Ang-2、Survivin的表达有关。（5）抗人IgM抗体抑制人鼻咽癌HNE-1细胞裸鼠移植瘤生长的机制可能与其促进凋亡、抑制细胞增殖和血管生成有关。
[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.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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