miRNA-9在骨肉瘤MG-63细胞增殖、凋亡中的作用及与CXCR4相关性研究

发布时间：2018-05-10 10:23

本文选题：骨肉瘤 + miRNA-9　；参考：《右江民族医学院》2017年硕士论文

【摘要】：目的:研究miRNA-9在骨肉瘤MG-63细胞中的作用,通过提高骨肉瘤MG-63细胞miRNA-9的表达,探讨miRNA-9在MG-63细胞增殖及凋亡中的作用及与CXCR4的相关性,为骨肉瘤的基因治疗提供理论依据。方法:骨肉瘤MG-63细胞随机分为miRNA-9转染组、阴性对照组和空白组。miRNA-9转染组转染miRNA-9(Oligo),阴性对照组转染阴性对照核苷酸序列(microRNA negative control sequence),空白组不做转染。荧光显微镜下观察细胞的转染效果,qRT-PCR检测细胞miRNA-9表达确定转染效果。CCK-8法检测3组细胞的增殖;流式细胞术比较3组细胞的凋亡水平;qRT-PCR检测各组细胞CXCR4 mRNA的表达水平。结果:(1)转染miRNA-9 Oligo后在荧光显微镜下能观察到绿色荧光,转染率达到70%,表明miRNA-9转染成功;(2)qRT-PCR结果:与阴性对照组和空白组相比,转染组细胞中miRNA-9表达升高(P0.01);(3)miRNA-9高表达的骨肉瘤细胞增殖速率在转染后24h、48h、72h、96h均较对照组和空白组降低(P均小于0.01),转染后24h细胞凋亡率增加(P0.01);(4)qRT-PCR结果显示miRNA-9高表达的骨肉瘤细胞CXCR4 mRNA表达水平下调(P0.01)。结论:(1)miRNA-9高表达能抑制骨肉瘤MG-63细胞细胞增殖,促进细胞凋亡。(2)CXCR4在骨肉瘤MG-63细胞中表达,CXCR4 mRNA与miRNA-9在骨肉瘤MG-63细胞中的表达水平呈负相关,即高表达miRNA-9的MG-63细胞低表达CXCR4 mRNA。CXCR4基因可能参与miRNA-9抑制骨肉瘤MG-63细胞的增殖,同时促进细胞凋亡。
[Abstract]:Objective: to study the role of miRNA-9 in osteosarcoma MG-63 cells, and to explore the role of miRNA-9 in MG-63 cell proliferation and apoptosis and its correlation with CXCR4 by increasing the expression of miRNA-9 in osteosarcoma MG-63 cells so as to provide a theoretical basis for gene therapy of osteosarcoma. Methods: osteosarcoma MG-63 cells were randomly divided into miRNA-9 transfection group, negative control group and blank group. MiRNA-9 transfection group was transfected with miRNA-9 oligodeoxynucleotide sequence, negative control group was transfected with microRNA negative control sequence, and blank group was not transfected. The effect of cell transfection was observed under fluorescence microscope. The expression of miRNA-9 was determined by qRT-PCR. The proliferation of the three groups was detected by CCK-8 method. The apoptosis level of the three groups was compared by flow cytometry and the expression of CXCR4 mRNA was detected by qRT-PCR. Results after transfection of miRNA-9 Oligo, green fluorescence could be observed under fluorescence microscope, and the transfection rate was 70%, which indicated that the successful transfection of miRNA-9 was compared with the negative control group and the blank group. The proliferation rate of osteosarcoma cells with high expression of P0.01MRNA-9 in the transfected group was significantly lower than that in the control group and blank group at 24 h after transfection for 48 h and 72 h / 96 h, respectively. The apoptosis rate of osteosarcoma cells was increased 24 h after transfection. The results of RT-PCR showed that miRNA-9 overexpression of osteosarcoma cells was higher than that of the control group and the blank group (P < 0.01). The results of RT-PCR showed that the proliferation rate of osteosarcoma cells with high expression of miRNA-9 was higher than that of control group and blank group. The expression of CXCR4 mRNA was down-regulated by P0.01. Conclusion the high expression of miRNA-9 can inhibit the proliferation of osteosarcoma MG-63 cells and promote the expression of CXCR4 mRNA in osteosarcoma MG-63 cells. There is a negative correlation between the expression of CXCR4 mRNA in osteosarcoma MG-63 cells and the expression of miRNA-9 in osteosarcoma MG-63 cells. The low expression of CXCR4 mRNA.CXCR4 gene in MG-63 cells with high expression of miRNA-9 may be involved in miRNA-9 inhibiting the proliferation of osteosarcoma MG-63 cells and promoting apoptosis.
【学位授予单位】：右江民族医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R738.1

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