SiRNA敲低USP7对喉癌细胞生物学特性的影响

发布时间：2018-03-16 04:31

本文选题：喉癌　切入点：USP7　出处：《川北医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:泛素特异性蛋白酶7(ubiquitin-specific protease 7,USP7)在肿瘤发生发展过程中有重要作用,其与喉癌(Laryngeal cancer)病理过程间的关系目前并不完全清楚。本研究通过小干扰RNA敲低USP7在喉癌细胞中的表达,以探讨USP7对喉癌细胞生物学特性的影响。方法:采用免疫组织化学Envision法检测73例喉癌组织和52例非癌组织中USP7蛋白的表达。进而设计并使用高效Si RNA(small interfering RNA)在喉癌HEP2细胞株特异性敲低USP7表达,然后利用CCK-8法、transwell小室迁移实验和流式细胞术观察USP7敲低后对喉癌细胞增殖、迁移能力和凋亡的影响。结果:USP7蛋白阳性表达率在喉癌组织和癌旁组织中分别为83.6%和36.5%,癌组织阳性表达率显著高于癌旁组织(χ2=29.1446,p0.0001);USP7蛋白阳性表达与肿瘤分化程度相关(χ2=3.9122,p=0.0479):肿瘤分化程度越低,USP7蛋白阳性表达率越高;在临床分期中,显示出T分期越晚,USP7阳性表达率越高,但差异没有统计学意义;自行设计的Si RNA可高效抑制喉癌HEP2细胞USP7蛋白m RNA表达,并利用最高效的2#Si RNA特异性敲低USP7蛋白表达,显著抑制了喉癌细胞的增殖、迁移能力,并促进了喉癌细胞的凋亡。结论:USP7在喉癌中高表达,与肿瘤分化程度成负相关;Si RNA-USP7可以显著抑制喉癌细胞增殖和迁移能力,并促进凋亡。因而,USP7在喉癌发生发展过程中扮演着重要角色,USP7可能是与喉癌诊断、治疗和预后有密切关系的一个重要生物学标志物。
[Abstract]:Objective: ubiquitin specific protease 7 (ubiquitin specific protease 7 USP7) plays an important role in the carcinogenesis and development of laryngeal carcinoma, and the relationship between it and the pathological process of laryngeal cancer is not completely clear. The present study aims to reduce the expression of USP7 in laryngeal carcinoma cells by small interfering RNA knockout. To investigate the effect of USP7 on the biological characteristics of laryngeal carcinoma cells. Methods: the expression of USP7 protein in 73 cases of laryngeal carcinoma and 52 cases of non-cancerous tissues was detected by immunohistochemical Envision method. Then high efficient Si RNA(small interfering was designed and used in laryngeal carcinoma. HEP2 cell line specifically knocked down the expression of USP7. Then the transwell chamber migration assay and flow cytometry were used to observe the proliferation of laryngeal carcinoma cells after USP7 knockout. Results the positive expression rates of 7% USP7 protein were 83.6% and 36.5 in laryngeal carcinoma and paracancerous tissues, respectively. The positive expression rate in cancer tissue was significantly higher than that in paracancerous tissue (蠂 2 29.1446p 0.0001) and the degree of tumor differentiation was correlated with the positive expression of USP7 protein. (蠂 ~ 2 / 3.9122 / P ~ (0.0479)): the lower the differentiation of tumor, the higher the positive expression rate of USP7 protein; In clinical stage, the higher the positive expression rate of USP7 7 was, the higher the positive rate of T stage was, but the difference was not statistically significant. The self-designed Si RNA could effectively inhibit the expression of USP7 protein m RNA in HEP2 cells of laryngeal carcinoma. The expression of 2 #Si RNA specific knockout protein was used to inhibit the proliferation and migration of laryngeal carcinoma cells and promote the apoptosis of laryngeal cancer cells. There is a negative correlation between tumor differentiation and tumor differentiation. Si RNA-USP7 can significantly inhibit the proliferation and migration ability of laryngeal cancer cells and promote apoptosis. Therefore, USP7 may play an important role in the development of laryngeal carcinoma and may be associated with the diagnosis of laryngeal carcinoma. Treatment and prognosis are closely related to an important biological marker.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R739.65

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