miR-24-3p在骨肉瘤中的表达及其抑制骨肉瘤细胞增殖和侵袭迁移的实验研究

miR-24-3p expressed in osteosarcoma and inhibited the proliferation,invasion and migration of osteosarcoma cells

目的:探讨miR-24-3p在骨肉瘤中的表达及调控骨肉瘤细胞生物学行为的作用。方法:收集我院骨肉瘤标本及配对癌旁组织标本10例,RT-qPCR检测骨肉瘤组织及癌旁组织中miR-24-3p的相对表达量。MG-63细胞转染miR-24-3p mimics或miR-24-3p inhibitor后,RT-qPCR检测miR-24-3p干扰及过表达效率。通过CCK-8及EdU实验评估骨肉瘤细胞增殖能力,划痕实验和Transwell实验检测骨肉瘤细胞迁移与侵袭能力。Western blot检测PCNA蛋白表达,流式细胞术检测细胞凋亡变化。结果:与癌旁组织相比,骨肉瘤组织及细胞中miR-24-3p表达显著下调。敲降miR-24-3p表达后,CCK-8及EdU实验结果表明MG-63细胞增殖能力增加,划痕实验和Tranwell实验结果显示MG-63细胞迁移与侵袭能力增加,流式细胞术结果显示细胞凋亡降低(P<0.001)。过表达miR-24-3p表达后,CCK-8、划痕实验和Tranwell实验结果显示MG-63细胞增殖、迁移与侵袭能力减弱,细胞凋亡增加(P<0.01)。Western blot结果显示miR-24-3p mimics组PCNA蛋白表达降低,miR-24-3p inhibitor组PCNA蛋白表达增加。结论:miR-24-3p在骨肉瘤组织及细胞中表达下调,并且可以抑制骨肉瘤细胞增殖、迁移与侵袭,促进细胞凋亡,发挥抑癌作用,有望成为骨肉瘤治疗的靶点之一。

Objective:To investigate the expression of miR-24-3p in osteosarcoma and the role of regulating the biological behavior of osteosarcoma cells.Methods:Ten osteosarcoma specimens and paired para-cancer tissue specimens from our hospital were collected.RT-qPCR was used to detect the relative expression of miR-24-3p in osteosarcoma and para-cancer tissues.MG-63 cells were transfected with miR-24-3p mimics or miR-24-3p inhibitor,and RT-qPCR detected the interference and overexpression efficiency.The proliferation of osteosarcoma cells was evaluated by CCK-8 and EdU assays,and the migration and invasion were assessed by scratch assay and Transwell assay.The expression of PCNA protein was detected by Western blot.Cell apoptosis was detected by flow cytometry.Results:The expression of miR-24-3p in osteosarcoma tissues and cells was significantly down-regulated by RT-qPCR.The proliferation of osteosarcoma cells was increased by CCK-8 and EdU assays when the miR-24-3p expression was down-regulated.The migration and invasion ability of osteosarcoma cells was increased by scratch and Transwell assay,while cell apoptosis was decreased by flow cytometry(P<0.001).The proliferation of osteosarcoma cells was inhibited by CCK-8 and EdU experiments when the miR-24-3p expression was up-regulated.The migration and invasion of osteosarcoma cells were inhibited by scratch and Transwell experiments after miR-24-3p overexpression,while cell apoptosis was increased(P<0.01).Western blot results showed that the expression of PCNA protein in the miR-24-3p mimics group was decreased,while that in the miR-24-3p inhibitor group was increased.Conclusion:miR-24-3p is downregulated in osteosarcoma tissues and cells and could inhibit the proliferation,migration and invasion of osteosarcoma cells and promote cell apoptosis.miR-24-3p plays a role in osteosarcoma inhibition and is expected to be one of the therapeutic targets for osteosarcoma patients.