本文選題：neuritin + 基因過(guò)表達(dá)　； 參考：《石河子大學(xué)》2007年碩士論文

【摘要】： 目的:(1)檢測(cè)部分腫瘤及相應(yīng)正常組織中neuritin的表達(dá)情況;(2)觀(guān)察neuritin過(guò)表達(dá)對(duì)細(xì)胞生物學(xué)特性的影響,探討n(yōu)euritin與腫瘤發(fā)生發(fā)展的相關(guān)性。 方法:(1)運(yùn)用免疫組織化學(xué)技術(shù)分別檢測(cè)5例肺癌、食管癌、子宮內(nèi)膜癌、宮頸癌及相應(yīng)正常組織中neuritin蛋白的表達(dá)情況。初步篩選出neuritin蛋白表達(dá)有差異的組織后,擴(kuò)大其樣本量利用免疫組化技術(shù)核實(shí)neuritin在腫瘤組織與正常組織的表達(dá)差異,并通過(guò)RT-PCR在新鮮組織中進(jìn)一步驗(yàn)證免疫組織化學(xué)檢測(cè)的結(jié)果。(2)利用穩(wěn)定表達(dá)neuritin的BMSCs細(xì)胞,通過(guò)β-半乳糖苷酶檢測(cè)觀(guān)察neuritin對(duì)細(xì)胞衰老的影響; Annexin V-PI染色后,經(jīng)通過(guò)流式細(xì)胞儀觀(guān)察neuritin對(duì)細(xì)胞凋亡的影響。 結(jié)果:(1)在上述組織中,免疫組化初步篩選結(jié)果顯示,與相應(yīng)正常組織相比,neuritin蛋白在子宮內(nèi)膜癌、宮頸癌、食管癌組織中高表達(dá);尤以肺癌與正常肺組織neuritin蛋白表達(dá)差異明顯;進(jìn)一步擴(kuò)大肺癌及正常肺組織樣本量,免疫組織化學(xué)結(jié)果顯示,肺癌組織中neuritin蛋白表達(dá)高于正常肺組織,差異有統(tǒng)計(jì)學(xué)意義(p0.01)。RT-PCR結(jié)果顯示,neuritin基因在肺癌組織有較高表達(dá),而在正常肺組織中未檢測(cè)到neuritin基因的表達(dá)。(2)β-半乳糖苷酶檢測(cè)結(jié)果顯示,neuritin穩(wěn)定過(guò)表達(dá)的骨髓間充質(zhì)干細(xì)胞(BMSc-neu)陽(yáng)性染色率(19.33±1.53%)低于陰性對(duì)照組(BMSc-3.1)陽(yáng)性染色率(47.67±3.05%),差異有統(tǒng)計(jì)學(xué)意義(t=12.14,p0.05);(3)Annexin V-PI染色結(jié)果顯示;neuritin穩(wěn)定過(guò)表達(dá)的骨髓間充質(zhì)干細(xì)胞(BMSc-neu)早期凋亡率(25.11±1.17%)低于陰性對(duì)照組(BMSc-3.1)早期凋亡率(38.10±1.28%),差異有統(tǒng)計(jì)學(xué)意義(t =9.22,p0.05)。 結(jié)論:(1)neuritin在部分腫瘤組織的表達(dá)高于其相應(yīng)的正常組織;(2)neuritin過(guò)表達(dá)能夠延緩BMSCs細(xì)胞的衰老;(3)neuritin過(guò)表達(dá)能抑制BMSCs細(xì)胞凋亡。 以上研究結(jié)果提示:neuritin作為抑制細(xì)胞凋亡的蛋白因子,可能與腫瘤的發(fā)生發(fā)展相關(guān)。
[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 鹵1.53) and the positive staining rate of BMSc-3.1) was lower than that of negative control group (47.67 鹵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 鹵1.17), which was significantly lower than that of negative control group (38.10 鹵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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R341

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