circ_0001246调节miRNA30a对骨肉瘤化疗敏感性影响的机制研究
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江苏省自然科学青年基金(BK20171092)


The mechanism of effect of circ_0001246 on osteosarcoma chemotherapy sensitivity by regulating miRNA31a
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    摘要:

    目的:探讨circ_0001246调节miRNA30a及影响骨肉瘤化疗敏感性的机制。方法:将骨肉瘤细胞随机分为3组,分别置于6孔板中,包括未转染组(空白组)、siRNA阴性对照转染组(NC组)和siRNA转染组(circ_0001246 siRNA转染组),转染后荧光倒置显微镜观察转染效率,检测各组细胞circ_0001246、miRNA30a的表达,用流式细胞AnnexinV/7AAD双染法检测各组细胞对化疗药物依托泊苷(etoposide -VP16)的敏感性,荧光素酶报告基因实验验证circ_0001246对miRNA30a的“吸附”作用。结果:circ_0001246 siRNA转染组的细胞miRNA30a表达较NC组和空白组明显上调(P < 0.05),骨肉瘤中circ_0001246和miRNA30a的表达呈现负反馈。circ_0001246 siRNA转染组细胞凋亡率较NC组和空白组明显升高(P < 0.05)。circ_0001246可以通过直接碱基配对吸附miRNA30a发挥调控作用。结论:circ_0001246是骨肉瘤相关癌基因,靶向抑制骨肉瘤中circ_0001246表达可以增强肿瘤细胞对化疗药物依托泊苷的敏感性,该作用可能是通过circ_0001246的吸附功能负调控miRNA30a表达而实现的。

    Abstract:

    Objective:This study aims to investigate the molecular mechanism of osteosarcoma chemotherapy sensitivity regulation of circ_0001246 via targeting miRNA30a. Methods:The MG63 and U2OS cells were randomly allocated into three groups in 6-well plates respectively,including non-transfection group(blank group), siRNA negative control transfection group(NC group),and siRNA transfection group(circ_0001246 siRNA). The transfection efficiencies were evaluated by fluorescence inverted microscope. Expression Level of circ_0001246 and miRNA30a were detected. Osteosarcoma chemotherapy sensitivity were detected by annexin V-FITC/7AAD apoptosis double staining. Function of circ_0001246 to adsorb miRNA30a were verified by luciferase reporter gene assay. Results: MiRNA30a expression in the circ_0001246 siRNA transfection group were significantly up-regulated compared to the NC group and blank group(P < 0.05). We found circ_0001246 and miRNA30a were inversely expressed. Osteosarcoma chemotherapy sensitivity in the circ_0001246 siRNA transfection group were significantly up-regulated compared to the NC group and blank group(P < 0.05). circ_0001246 could adsorb miRNA30a by direct base pairs. Conclusion: Circ_0001246 is an osteosarcoma related oncogene. Targeted inhibition of circ_0001246 expression in osteosarcoma can enhance the chemotherapy sensitivity of tumor cells to etoposide-VP16. One of the mechanisms is the negative regulation of the miRNA30a expression by the adsorption function of circ_0001246.

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王 娟,吴 涛,何 斌,王伯尧,魏新程,范 磊. circ_0001246调节miRNA30a对骨肉瘤化疗敏感性影响的机制研究[J].南京医科大学学报(自然科学版),2021,(2):198-202

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  • 收稿日期:2020-04-30
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  • 在线发布日期: 2021-03-10
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