本文選題：骨肉瘤 + miRNA-9��； 參考：《右江民族醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:研究miRNA-9在骨肉瘤MG-63細(xì)胞中的作用,通過提高骨肉瘤MG-63細(xì)胞miRNA-9的表達(dá),探討miRNA-9在MG-63細(xì)胞增殖及凋亡中的作用及與CXCR4的相關(guān)性,為骨肉瘤的基因治療提供理論依據(jù)。方法:骨肉瘤MG-63細(xì)胞隨機(jī)分為miRNA-9轉(zhuǎn)染組、陰性對(duì)照組和空白組。miRNA-9轉(zhuǎn)染組轉(zhuǎn)染miRNA-9(Oligo),陰性對(duì)照組轉(zhuǎn)染陰性對(duì)照核苷酸序列(microRNA negative control sequence),空白組不做轉(zhuǎn)染。熒光顯微鏡下觀察細(xì)胞的轉(zhuǎn)染效果,qRT-PCR檢測(cè)細(xì)胞miRNA-9表達(dá)確定轉(zhuǎn)染效果。CCK-8法檢測(cè)3組細(xì)胞的增殖;流式細(xì)胞術(shù)比較3組細(xì)胞的凋亡水平;qRT-PCR檢測(cè)各組細(xì)胞CXCR4 mRNA的表達(dá)水平。結(jié)果:(1)轉(zhuǎn)染miRNA-9 Oligo后在熒光顯微鏡下能觀察到綠色熒光,轉(zhuǎn)染率達(dá)到70%,表明miRNA-9轉(zhuǎn)染成功;(2)qRT-PCR結(jié)果:與陰性對(duì)照組和空白組相比,轉(zhuǎn)染組細(xì)胞中miRNA-9表達(dá)升高(P0.01);(3)miRNA-9高表達(dá)的骨肉瘤細(xì)胞增殖速率在轉(zhuǎn)染后24h、48h、72h、96h均較對(duì)照組和空白組降低(P均小于0.01),轉(zhuǎn)染后24h細(xì)胞凋亡率增加(P0.01);(4)qRT-PCR結(jié)果顯示miRNA-9高表達(dá)的骨肉瘤細(xì)胞CXCR4 mRNA表達(dá)水平下調(diào)(P0.01)。結(jié)論:(1)miRNA-9高表達(dá)能抑制骨肉瘤MG-63細(xì)胞細(xì)胞增殖,促進(jìn)細(xì)胞凋亡。(2)CXCR4在骨肉瘤MG-63細(xì)胞中表達(dá),CXCR4 mRNA與miRNA-9在骨肉瘤MG-63細(xì)胞中的表達(dá)水平呈負(fù)相關(guān),即高表達(dá)miRNA-9的MG-63細(xì)胞低表達(dá)CXCR4 mRNA。CXCR4基因可能參與miRNA-9抑制骨肉瘤MG-63細(xì)胞的增殖,同時(shí)促進(jìn)細(xì)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.
【學(xué)位授予單位】：右江民族醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R738.1

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