人部分组织中neuritin表达检测及其过表达对细胞凋亡的影响

发布时间：2018-04-21 16:50

本文选题：neuritin + 基因过表达　；参考：《石河子大学》2007年硕士论文

【摘要】： 目的:(1)检测部分肿瘤及相应正常组织中neuritin的表达情况;(2)观察neuritin过表达对细胞生物学特性的影响,探讨neuritin与肿瘤发生发展的相关性。方法:(1)运用免疫组织化学技术分别检测5例肺癌、食管癌、子宫内膜癌、宫颈癌及相应正常组织中neuritin蛋白的表达情况。初步筛选出neuritin蛋白表达有差异的组织后,扩大其样本量利用免疫组化技术核实neuritin在肿瘤组织与正常组织的表达差异,并通过RT-PCR在新鲜组织中进一步验证免疫组织化学检测的结果。(2)利用稳定表达neuritin的BMSCs细胞,通过β-半乳糖苷酶检测观察neuritin对细胞衰老的影响; Annexin V-PI染色后,经通过流式细胞仪观察neuritin对细胞凋亡的影响。结果:(1)在上述组织中,免疫组化初步筛选结果显示,与相应正常组织相比,neuritin蛋白在子宫内膜癌、宫颈癌、食管癌组织中高表达;尤以肺癌与正常肺组织neuritin蛋白表达差异明显;进一步扩大肺癌及正常肺组织样本量,免疫组织化学结果显示,肺癌组织中neuritin蛋白表达高于正常肺组织,差异有统计学意义(p0.01)。RT-PCR结果显示,neuritin基因在肺癌组织有较高表达,而在正常肺组织中未检测到neuritin基因的表达。(2)β-半乳糖苷酶检测结果显示,neuritin稳定过表达的骨髓间充质干细胞(BMSc-neu)阳性染色率(19.33±1.53%)低于阴性对照组(BMSc-3.1)阳性染色率(47.67±3.05%),差异有统计学意义(t=12.14,p0.05);(3)Annexin V-PI染色结果显示;neuritin稳定过表达的骨髓间充质干细胞(BMSc-neu)早期凋亡率(25.11±1.17%)低于阴性对照组(BMSc-3.1)早期凋亡率(38.10±1.28%),差异有统计学意义(t =9.22,p0.05)。结论:(1)neuritin在部分肿瘤组织的表达高于其相应的正常组织;(2)neuritin过表达能够延缓BMSCs细胞的衰老;(3)neuritin过表达能抑制BMSCs细胞凋亡。以上研究结果提示:neuritin作为抑制细胞凋亡的蛋白因子,可能与肿瘤的发生发展相关。
[Abstract]:Objective to detect the expression of neuritin in some tumors and normal tissues. To investigate the effect of neuritin overexpression on cell biological characteristics and to explore the relationship between neuritin and tumorigenesis and development. Methods the expression of neuritin protein in 5 cases of lung cancer, esophageal carcinoma, endometrial carcinoma, cervical carcinoma and corresponding normal tissues were detected by immunohistochemistry. After preliminary screening out the tissues with different expression of neuritin protein, we expanded its sample size to verify the difference of neuritin expression between tumor tissues and normal tissues by immunohistochemical technique. The results of immunohistochemistry were further verified by RT-PCR in fresh tissues. The BMSCs cells expressing neuritin stably were used to observe the effect of neuritin on cell senescence by 尾 -galactosidase assay. After Annexin V-PI staining, the effect of neuritin on cell senescence was observed by 尾 -galactosidase assay. The effect of neuritin on apoptosis was observed by flow cytometry. Results compared with the normal tissues, the expression of neuritin protein was higher in endometrial carcinoma, cervical cancer and esophageal carcinoma, especially in lung cancer and normal lung tissue. The immunohistochemical results showed that the expression of neuritin protein in lung cancer tissues was higher than that in normal lung tissues, and the difference was statistically significant (p 0.01). RT-PCR showed that the expression of neuritin gene in lung cancer tissues was higher than that in normal lung tissues. In normal lung tissues, the expression of neuritin gene was not detected. 2) 尾 -galactosidase assay showed that the positive staining rate of bone marrow mesenchymal stem cells with stable expression of neuritin was 19.33 卤1.53) and the positive staining rate of BMSc-3.1) was lower than that of negative control group (47.67 卤3.05). The results of Annexin V-PI staining showed that the early apoptosis rate of bone marrow mesenchymal stem cells (BMSc-neu) with stable expression of neuritin was 25.11 卤1.17), which was significantly lower than that of negative control group (38.10 卤1.28p0.05). Conclusion the overexpression of neuritin in some tumor tissues is higher than that in normal tissues. It can delay the senescence of BMSCs cells and inhibit the apoptosis of BMSCs cells. The above results suggest that as a protein factor that inhibits apoptosis, it may be related to the occurrence and development of tumor.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R341

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