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微小RNA-34a通过影响细胞凋亡抑制蛋白2(C-IAP2)和Bcl-2,部分抑制骨肉瘤的肿瘤侵袭和转移。

MicroRNA-34a inhibits tumor invasion and metastasis in osteosarcoma partly by effecting C-IAP2 and Bcl-2.

作者信息

Wen Jie, Zhao Yan-Kun, Liu Yan, Zhao Jin-Feng

机构信息

1 Department of Orthopedics, Xiangya Hospital, Central South University, Changsha, China.

2 Department of Orthopedics, Inner mongolia Baogang Hospital, The Third Affiliated Hospital of Inner Mongolia Medical College, Baotou, China.

出版信息

Tumour Biol. 2017 Jun;39(6):1010428317705761. doi: 10.1177/1010428317705761.

Abstract

Osteosarcoma is a common primary malignant bone tumor that occurs mainly in children and adolescents. Recent evidence has demonstrated that miR-34a is involved in the invasion and metastasis of osteosarcoma. This study aims to explore the effect of biological behavior of miR-34a on osteosarcoma. First, we collect osteosarcoma and adjacent specimens, and the relative expression of miR-34a and C-IAP2 messenger RNA was quantitated by real-time polymerase chain reaction. Furthermore, miR-34a stimulant is synthesized and transfected onto osteosarcoma MG-63 cells. The effect of overexpression of miR-34a on osteosarcoma was detected by colony-forming assay, Annexin V-fluorescein isothiocyanate Apoptosis Detection Kit I, Transwell assay, and animal experiment in vivo. Finally, the relative levels of C-IAP2 and Bcl-2 protein were checked by western blot, and the activity of caspase-3 and caspase-9 was tested by spectrophotometry assay. In conclusion, miR-34a was downregulated in osteosarcoma cells. And the expression of C-IAP2 and Bcl-2 protein was drastically inhibited, and the activities of caspase-3 and caspase-9 were significantly increased after transfecting miR-34a onto osteosarcoma MG-63 cells. And the overexpression of miR-34a can inhibit cell invasion and metastasis, promote cell apoptosis, and arrest cells in G0/G1 period. And the animal experiment in vivo demonstrated that the overexpression of miR-34a could significantly inhibit the growth of osteosarcoma in animal skin. Taken together, we indicated that miR-34a can inhibit tumor invasion and metastasis in osteosarcoma, and its mechanism may be partly related to downregulating the expression of C-IAP2 and Bcl-2 protein directly or indirectly.

摘要

骨肉瘤是一种常见的原发性恶性骨肿瘤,主要发生于儿童和青少年。最近的证据表明,miR-34a参与了骨肉瘤的侵袭和转移。本研究旨在探讨miR-34a对骨肉瘤生物学行为的影响。首先,我们收集骨肉瘤及相邻标本,通过实时聚合酶链反应定量检测miR-34a和C-IAP2信使核糖核酸的相对表达。此外,合成miR-34a激动剂并转染到骨肉瘤MG-63细胞上。通过集落形成试验、膜联蛋白V-异硫氰酸荧光素凋亡检测试剂盒I、Transwell试验和体内动物实验检测miR-34a过表达对骨肉瘤的影响。最后,通过蛋白质免疫印迹法检测C-IAP2和Bcl-2蛋白的相对水平,通过分光光度法检测半胱天冬酶-3和半胱天冬酶-9的活性。总之,miR-34a在骨肉瘤细胞中表达下调。将miR-34a转染到骨肉瘤MG-63细胞后,C-IAP2和Bcl-2蛋白的表达受到显著抑制,半胱天冬酶-3和半胱天冬酶-9的活性显著增加。miR-34a的过表达可抑制细胞侵袭和转移、促进细胞凋亡,并使细胞停滞于G0/G1期。体内动物实验表明,miR-34a的过表达可显著抑制动物皮肤中骨肉瘤的生长。综上所述,我们表明miR-34a可抑制骨肉瘤的肿瘤侵袭和转移,其机制可能部分与直接或间接下调C-IAP2和Bcl-2蛋白的表达有关。

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