弥漫大B细胞淋巴瘤中miR-29b的表达及功能研究

发布时间：2018-04-26 19:21

本文选题：弥漫大B细胞淋巴瘤 + miR-29b　；参考：《郑州大学》2015年硕士论文

【摘要】：弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)是一组临床和生物学特性各异的异质性肿瘤,约占非霍奇金淋巴瘤(non-hodgkin’s lymphoma,NHL)的40%,其在发展中国家高达60%,而现有治疗方法只能使40-50%的病人获得临床持续缓解。因此寻找新的分子标志在临床上对其诊断和治疗有着一定的积极意义。Micro RNAs(mi RNAs)作为一种单链的非编码小分子RNA,其长度约为22-25个碱基对构成,该类RNA在转录之后对基因的表达起到调控作用,根据抑制或是降解靶基因功能其分为两种。研究表明,mi RNAs对免疫系统的调控,在体内细胞的增殖分化和生长发育起着一定的重要作用,对肿瘤的发生发展也发挥着一定的抑制或促进作用。在不同的肿瘤中,不同的mi RNAs表达模式也不尽相同,因此在临床上可作为预测、诊断以及治疗的生物学标志。研究表明mi R-29b作为mi RNAs中的一员,它可以沉默一些潜在的癌基因,抑制肿瘤细胞生长,诱导凋亡,发挥抑癌基因的作用。研究也表明,mi R-29b功能存在着异质性,目前mi R-29b在DLBCL中的表达及功能还缺乏相应的研究,因此本研究通过分析mi R-29b在DLBCL细胞系中的表达、功能,探讨mi R-29b在DLBCL发生发展中的功能。目的了解DLBCL细胞系中mi R-29b的表达情况;观察mi R-29b mimic和mi R-29b inhibitor在DLBCL两种细胞系SU-DHL-8,SU-DHL-10的转染率情况;探讨mi R-29b对SU-DHL-8,SU-DHL-10两种细胞系增殖情况的影响;研究mi R-29b对DLBCL两种细胞系SU-DHL-8,SU-DHL-10细胞迁移能力的影响。研究mi R-29b对DLBCL两种细胞系SU-DHL-8,SU-DHL-10细胞凋亡的影响。方法1、利用Trizol试剂提取淋巴细胞和人正常淋巴结组织中总RNA,根据mi R-29b设计的反转录引物及q RT-PCR引物;利用q RT-PCR方法检测DLBCL细胞系中mi R-29b的表达情况。2、根据mi R-29b的序列合成mi R-29b mimic和mi R-29b inhibitor,将合成的mi R-29b mimic和mi R-29b inhibitor转染入DLBCL细胞系中,通过q RT-PCR方法检测mi R-29b的表达量。3、通过利用CCK-8细胞生长实验检测mi R-29b对DLBCL细胞系细胞增殖能力的影响;4、运用Transwell迁移实验检测mi R-29b对DLBCL细胞系细胞迁移能力的影响,并通过流式细胞仪检测细胞的凋亡情况。结果1.通过q RT-PCR方法检测DLBCL细胞系中mi R-29b的表达结果显示,mi R-29b在人正常淋巴结组织中表达显著高于SU-DHL-10和SU-DHL-8细胞系中的表达量(P0.05)。2.mi R-29b mimic和mi R-29b inhibitor在DLBCL细胞系中转染效率结果显示,随着浓度的改变,mi R-29b inhibitor转染的细胞数量变化不明显,差异没有统计学意义(P0.05);而随着浓度的增加,mi R-29b mimic转染的细胞数量显著增加(P0.05)。3.分别转染mi R-29b mimic和mi R-29b inhibitor之后可改变DLBCL细胞mi R-29b的表达,和正常mi R-29b的表达量相比,mi R-29b mimic的转染使mi R-29b的表达量提高了200-300倍,结果有显著性差异(P0.05);mi R-29b inhibitor和对照组相比抑制了mi R-29b的表达,使mi R-29b的表达量降低50%左右(P0.05)。4.采用CCK-8细胞生长实验检测mi R-29b对SU-DHL-8,SU-DHL-10细胞系增殖的影响。结果显示:和对照组相比,上调mi R-29b能够显著抑制细胞的增殖能力(P0.05),而下调mi R-29b则细胞的增殖能力显著增强(P0.05)。5.采用Transwell迁移实验检测mi R-29b对DLBCL细胞系细胞迁移能力的影响。结果显示:和对照组相比,上调mi R-29b可以显著抑制细胞迁移能力(P0.05),而下调mi R-29b则细胞的迁移能力显著增强(P0.05)。结论1.mi R-29b在DLBCL中的低表达,推测mi R-29b表达可能参与了弥漫大B细胞淋巴瘤的发生发展过程。2.mi R-29b在DLBCL中可以发挥着抑癌基因的作用,这有望成为新的以mi RNA为基础的肿瘤干预物,尚需更深入研究。
[Abstract]:Diffuse large B cell lymphoma (diffuse large B-cell lymphoma, DLBCL) is a heterogeneous group of heterogeneous tumors with different clinical and biological characteristics, accounting for 40% of non Hodgkin's lymphoma (non-Hodgkin 's lymphoma, NHL), which is up to 60% in developing countries, and the existing treatment methods can only make persistent remission in 40-50% patients. The new molecular markers have certain positive significance in the clinical diagnosis and treatment of.Micro RNAs (MI RNAs) as a single strand of non coding small molecule RNA, whose length is about 22-25 base pairs, and this class RNA plays a regulatory role in the expression of genes after transcription, and the root is suppressed or degrade target genes into two kinds. The study shows that MI RNAs plays an important role in the proliferation, differentiation and growth of the immune system in the body. It also plays a certain inhibitory or promoting role on the development and development of the tumor. In different tumors, different mi RNAs expression patterns are different, so it can be used as a prediction, diagnosis and treatment in clinical. The biological markers of therapy show that MI R-29b, as a member of MI RNAs, can silence some of the potential oncogenes, inhibit the growth of tumor cells, induce apoptosis and play the role of tumor suppressor gene. The study also shows that the MI R-29b function exists heterogeneity, and the expression and function of MI R-29b in DLBCL are still lack of corresponding research, therefore, the expression and function of MI R-29b in DLBCL are still lacking. In this study, the expression and function of MI R-29b in the DLBCL cell line were analyzed to explore the function of MI R-29b in the development of DLBCL. Objective to understand the expression of MI R-29b in the DLBCL cell line, and to observe the transfection rate of MI R-29b mimic and MI R-29b mimic. The effect of the proliferation of two -10 cell lines; the effect of MI R-29b on the migration of SU-DHL-8 and SU-DHL-10 cells in the DLBCL two cell lines. The effect of MI R-29b on the apoptosis of DLBCL two cell lines SU-DHL-8 and SU-DHL-10 cells. Method 1 The expression of MI R-29b in the DLBCL cell line was detected by the reverse transcription primers and Q RT-PCR primers, and the MI R-29b expression in the DLBCL cell line was detected by the Q RT-PCR method. The effect of MI R-29b on the proliferation of DLBCL cell lines was detected by CCK-8 cell growth test. 4, the effect of MI R-29b on the cell migration ability of DLBCL cell lines was detected by Transwell migration test, and the cell apoptosis was detected by flow cytometry. Results the expression of MI expression in DLBCL cell line was detected by Q RT-PCR method. The results showed that the expression of MI R-29b in human normal lymph nodes was significantly higher than that in SU-DHL-10 and SU-DHL-8 cell lines (P0.05).2.mi R-29b mimic and MI R-29b inhibitor in the DLBCL cell lines. As the concentration increased, the number of cells transfected by Mi R-29b mimic increased significantly (P0.05).3. transfected to MI R-29b mimic and MI R-29b inhibitor, which could be changed by 200-300 times compared with the expression of normal mimic. There was significant difference (P0.05); MI R-29b inhibitor inhibited the expression of MI R-29b and reduced the expression of MI R-29b by about 50% (P0.05).4. (P0.05).4. using CCK-8 cell growth test to detect the effect of MI cell proliferation. Proliferation ability (P0.05), and down regulation of MI R-29b, the proliferation ability of cells increased significantly (P0.05).5. using Transwell migration test to detect the effect of MI R-29b on the migration ability of DLBCL cell line cells. The results showed that up regulation of MI R-29b could significantly inhibit cell migration capacity (P0.05), while down regulation, the migration energy of cells was down regulated. Force significantly enhanced (P0.05). Conclusion the low expression of 1.mi R-29b in DLBCL, it is presumed that MI R-29b expression may be involved in the development of diffuse large B cell lymphoma..2.mi R-29b can play the role of tumor suppressor gene in DLBCL, which is expected to be a new tumor intervention based on MI RNA. It is still needed to be further studied.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R733.1

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