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[微小RNA-181a通过靶向RASSF1A促进骨肉瘤细胞的增殖和转移]

[miR-181a promotes the proliferation and metastasis of osteosarcoma cells by targeting RASSF1A].

作者信息

Zang Xiaofang, Li Qin, Wang Weiguo, Zhou Yong, Chen Shijie, Xiao Tao

机构信息

Department of Orthopaedics, Third Xiangya Hospital, Central South University, Changsha 410013, China.

Department of Orthopaedics, Second Xiangya Hospital, Central South University, Changsha 410011, China.

出版信息

Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2016 Aug;41(8):789-95. doi: 10.11817/j.issn.1672-7347.2016.08.003.

Abstract

OBJECTIVE

To investigate the role of miR-181a in promoting the proliferation and metastasis of osteosarcoma cells by targeting RASSF1A.

METHODS

The level of miR-181a in 30 human osteosarcoma tissues and corresponding bone tissues was detected by real-time PCR, and the correlation between the level of miR-181a and clinicopathological characteristics of osteosarcoma was analyzed. Osteosarcoma cells MG-63 were transfected with chemically-synthesized miR-181a mimics and inhibitors, and the proliferation, migration and invasion of MG-63 cells were detected by MTT and Transwell assay. The specific binding ability of miR-181a to RASSF1A 3'-UTR was theoretically predicted and detected by the dual luciferase reporter gene assay.

RESULTS

The level of miR-181a in osteosarcoma tissues was statistically higher than that in the corresponding bone tissues (P<0.001). However, the level of miR-181a was not correlated with gender, age, tumor size and stage (P>0.05). MTT and Transwell assays showed that the growth rate, migration and invasion ability of MG-63 cells with up-regulation of miR-181a was significantly increased compared with negative control (P<0.05), while the growth rate, migration and invasion ability of MG-63 with down-regulated miR-181a was significantly decreased compared with negative control (P<0.05). Luciferase reporter gene assay showed that miR-181a targeted the 3'-UTR of RASSF1A and regulated the expression of RASSF1A.

CONCLUSION

MiR-181a promotes the proliferation and metastasis of osteosarcoma cells through specifically binding to RASSF1A 3'-UTR and subsequent down-regulation of RASSF1A.

摘要

目的

探讨miR-181a通过靶向RASSF1A促进骨肉瘤细胞增殖和转移的作用。

方法

采用实时PCR检测30例人骨肉瘤组织及相应骨组织中miR-181a的水平,并分析miR-181a水平与骨肉瘤临床病理特征的相关性。用化学合成的miR-181a模拟物和抑制剂转染骨肉瘤细胞MG-63,通过MTT和Transwell实验检测MG-63细胞的增殖、迁移和侵袭能力。通过双荧光素酶报告基因实验从理论上预测并检测miR-181a与RASSF1A 3'-UTR的特异性结合能力。

结果

骨肉瘤组织中miR-181a水平在统计学上高于相应骨组织(P<0.001)。然而,miR-181a水平与性别、年龄、肿瘤大小和分期无关(P>0.05)。MTT和Transwell实验表明,与阴性对照相比,miR-181a上调的MG-63细胞的生长速率、迁移和侵袭能力显著增加(P<0.05),而miR-181a下调的MG-63细胞的生长速率、迁移和侵袭能力与阴性对照相比显著降低(P<0.05)。荧光素酶报告基因实验表明,miR-181a靶向RASSF1A的3'-UTR并调节RASSF1A的表达。

结论

MiR-181a通过与RASSF1A 3'-UTR特异性结合并随后下调RASSF1A来促进骨肉瘤细胞的增殖和转移。

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