lncRNA LINC00173通过调节miR-130a-5p抑制骨肉瘤发生发展的机制研究

林浩; 徐德利; 张凯

doi:10.13898/j.cnki.issn.1000-2200.2022.12.002

lncRNA LINC00173通过调节miR-130a-5p抑制骨肉瘤发生发展的机制研究

Mechanism of lncRNA LINC00173 inhibiting the development of osteosarcoma by regulating miR-130a-5p

摘要

摘要:
目的探讨长链非编码RNA（lncRNA）LINC00173通过调控miR-130a-5p抑制骨肉瘤发生发展的分子机制。
方法体外培养人骨肉瘤细胞系MG-63，分别将LINC00173过表达载体质粒或空载质粒转染至MG-63细胞，实时荧光定量PCR（qPCR）检测转染效率，采用细胞计数法（CCK-8）和Transwell实验分析MG-63细胞增殖、侵袭和迁移能力，双荧光素酶报告基因实验和qPCR检测LINC00173和miR-130a-5p的靶向调控关系。同时过表达LINC00173和miR-130a-5p，并检测MG-63细胞增殖、侵袭和迁移能力。
结果qPCR检测结果显示，与Mock组比较，pc-LINC00173组LINC00173在MG-63细胞中的表达量明显升高（P < 0.01）。转染48 h和72 h后，pc-LINC00173组细胞增殖活力均低于Mock组和pcDNA组（P < 0.01）。Transwell检测结果显示，pc-LINC00173组与Mock组比较侵袭和迁移细胞数明显减少（P < 0.05）。LINC00173与miR-130a-5p之间存在特异性结合位点，与miR-NC组比较，转染miR-130a-5p mimics能够抑制LINC00173-Wt细胞的相对荧光素酶活性（P < 0.01），与Mock组和pcDNA组比较，pc-LINC00173组MG-63细胞中miR-130a-5p的表达量均降低（P < 0.01）。与pc-LINC00173组和pc-LINC00173+miR-NC组比较，pc-LINC00173+miR-130a-5p组MG-63细胞中miR-130a-5p的表达明显升高，吸光度值明显升高，侵袭和迁移细胞数明显增多（P < 0.01）。
结论LINC00173能够抑制骨肉瘤MG-63细胞增殖、侵袭和迁移，其作用机制可能与靶向抑制miR-130a-5p的表达有关。

Abstract:
ObjectiveTo investigate the molecular mechanism of long-chain non-coding RNA (lncRNA) LINC00173 through regulating miR-130a-5p to mediate the development of osteosarcoma.
MethodsHuman osteosarcoma cell line MG-63 was cultured in vitro, and LINC00173 overexpression vector plasmid or empty plasmid was transfected into MG-63 cells, real-time fluorescence quantitative PCR (qPCR) was used to detect the transfection efficiency. CCK-8 and Transwell assay was used to analyze the proliferation, invasion and migration ability of MG-63 cells, dual luciferase reporter gene experiment and qPCR were used to detect the targeted regulation relationship between LINC00173 and miR-130a-5p. At the same time, LINC00173 and miR-130a-5p were overexpressed, and the proliferation, invasion and migration ability of MG-63 cells were detect.
ResultsqPCR results showed that the expression of LINC00173 in MG-63 cells in pc-LINC00173 group was significantly higher than that in Mock group (P < 0.01). After 48 and 72 hours of transfection, the cell proliferation activity of pc-LINC00173 group was lower than that of Mock group and pcDNA group (P < 0.01). Transwell results showed that the number of invading and migrating cells in pc-LINC00173 group was significantly reduced compared with Mock group (P < 0.05). There was a specific binding site between LINC00173 and miR-130a-5p. Compared with the miR-NC group, transfection of miR-130a-5p mimics could inhibit the relative luciferase activity of LINC00173-Wt cells (P < 0.01). Compared with the Mock group and the pcDNA group, the expression of miR-130a-5p in MG-63 cells in pc-LINC0173 group decreased (P < 0.01). Compared with pc-LINC00173 group and pc-LINC00173 + miR-NC group, the expression of miR-130a-5p in MG-63 cells in pc-LINC00173 + miR-130a-5p group was significantly increased, the absorbance value was significantly increased, and the number of invading and migrating cells was significantly increased (P < 0.01).
ConclusionsLINC00173 can inhibit the proliferation, invasion and migration of osteosarcoma MG-63 cells, and its mechanism may be related to the targeted inhibition of miR-130a-5p expression.

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