KPNA2在骨肉瘤组织中的表达情况及对其骨肉瘤细胞系增殖影响的研究

发布时间：2018-05-22 11:35

本文选题：KPNA2 + 骨肉瘤　；参考：《山西医科大学》2017年硕士论文

【摘要】：背景:核转运蛋白α2是核转运蛋白的一种,在肿瘤的发生发展中起着重要的作用。研究表明KPNA2在多种肿瘤中表达,然而目前并无有关KPNA2在骨肉瘤中的作用相关报道。目的:本实验研究骨肉瘤细胞系中和正常组织中KPNA2的蛋白及m RNA的表达差异。并进一步研究KPNA2是否影响骨肉瘤细胞系Saos-2的增值和凋亡。方法:免疫组化(IHC)用于检测骨肉瘤组织和正常骨组织中KPNA2的蛋白的表达量;实时荧光定量PCR(q PCR)用于检测四种骨肉瘤细胞系中KPNA2m RNA表达水平,小片段RNA干扰慢病毒技术用于敲减Saos-2中KPNA2的表达;四唑盐比色法(MTT)和高内涵筛选(HCS)用于检测细胞的增值和生长;流式细胞术用于检测细胞周期和细胞凋亡。结果:结果显示骨肉瘤组织中KPNA2高表达,并且在四种骨肉瘤细胞系中KPNA2m RNA也高表达。在转染慢病毒后,与对照组相比,实验组中细胞增值率明显下降,流式结果显示实验组中在S期和G2/M期细胞数目明显增多,表明KPNA2可能干扰Saos-2的细胞周期和细胞的生长。细胞凋亡的结果表明KPNA2-sh RNA组的凋亡率为13.38±0.48%,明显高于对照组(5.13±0.33%)。结论:该实验表明,KPNA2在骨肉瘤中高表达,干扰骨肉瘤细胞中KPNA2的表达可抑制Saos-2细胞的增值,促进其凋亡。
[Abstract]:Background: nuclear transporter 伪 2 is a nuclear transporter, which plays an important role in tumorigenesis and development. Studies have shown that KPNA2 is expressed in a variety of tumors, but there are no reports on the role of KPNA2 in osteosarcoma. Objective: to study the expression of KPNA2 protein and m RNA in osteosarcoma cell line and normal tissue. Furthermore, the effect of KPNA2 on the proliferation and apoptosis of osteosarcoma cell line Saos-2 was studied. Methods: immunohistochemistry was used to detect the expression of KPNA2 protein in osteosarcoma and normal bone tissue, and real-time fluorescence quantitative PCR(q was used to detect the expression of KPNA2m RNA in four kinds of osteosarcoma cell lines. Small fragment RNA interference lentivirus technique was used to knock down the expression of KPNA2 in Saos-2; tetrazolium colorimetric assay (MTT) and high intension screening assay were used to detect cell proliferation and growth; flow cytometry was used to detect cell cycle and apoptosis. Results: high expression of KPNA2 was found in osteosarcoma tissues and high expression of KPNA2m RNA in four kinds of osteosarcoma cell lines. Compared with the control group, the proliferation rate of lentivirus in the experimental group was significantly decreased, and the number of cells in S phase and G 2 / M phase was significantly increased in the experimental group, indicating that KPNA2 might interfere with the cell cycle and cell growth of Saos-2. The apoptosis rate in KPNA2-sh RNA group was 13.38 卤0.48, which was significantly higher than that in control group (5.13 卤0.33). Conclusion: KPNA2 is highly expressed in osteosarcoma and interfering with the expression of KPNA2 in osteosarcoma cells can inhibit the proliferation of Saos-2 cells and promote its apoptosis.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R738.1

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