siRNA抑制骨肉瘤U2OS细胞系中SIRT6的表达及其生物学作用研究

doi:10.19401/j.cnki.1007-3639.2018.10.001

中国癌症杂志 ›› 2018, Vol. 28 ›› Issue (10): 721-725.doi: 10.19401/j.cnki.1007-3639.2018.10.001

siRNA抑制骨肉瘤U2OS细胞系中SIRT6的表达及其生物学作用研究

余　天¹，李　娟²，张伟凯³，张　帆³

1. 华中科技大学同济医学院附属同济医院眼科，湖北武汉 430030 ；
2. 华中科技大学同济医学院附属协和医院干细胞研究与应用中心，湖北武汉 430022 ；
3. 华中科技大学同济医学院附属同济医院骨科，湖北武汉 430030

出版日期:2018-10-30 发布日期:2018-11-12
通信作者: 张帆 E-mail: doctorzhphd@yeah.net
基金资助:
国家自然科学基金（81202093）。

Influence of down-regulation of SIRT6 expression by small interfering RNA on U2OS osteosarcoma cell line

YU Tian¹, LI Juan², ZHANG Weikai³, ZHANG Fan³

1. Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China; 2. Centre for Stem Cell Research and Application, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China; 3. Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China

Published:2018-10-30 Online:2018-11-12
Contact: ZHANG Fan E-mail: doctorzhphd@yeah.net

摘要/Abstract

摘要： 背景与目的：sirtuin6(SIRT6)蛋白是对人体细胞生长、代谢调控及应激反应等具有重要作用的一种核蛋白，常在细胞核中发挥多种调控作用。但该蛋白在骨肉瘤中的作用还不清楚，本研究通过下调人骨肉瘤U2OS细胞系中SIRT6的表达，观察其对U2OS细胞的作用。方法：设计合成SIRT6的siRNA序列，通过Lipofectamine^TM2000转染U2OS细胞系，通过蛋白质印迹法（Western blot）检测SIRT6蛋白的表达情况，利用流式细胞术检测细胞凋亡，细胞计数试剂盒（cellcounting kit-8，CCK-8）法检测细胞增殖，Transwell法检测细胞侵袭和迁移能力，CCK-8法分析对紫杉醇的敏感性。结果：成功构建SIRT6 siRNA序列，Western blot检测转染后24、48和72 h的蛋白水平，实验组较对照组明显降低。流式细胞术检测细胞凋亡也显示实验组凋亡增加（P＜0.05）。Transwell法检测结果显示，实验组侵袭与迁移能力较对照组下降（P＜0.05）。CCK-8法检测细胞生长情况，可见经紫杉醇处理后实验组生长较对照组下降明显，差异有统计学意义（P＜0.05）。结论：下调骨肉瘤U2OS细胞中SIRT6的表达可以抑制骨肉瘤细胞侵袭，促进凋亡，并增强骨肉瘤细胞对紫杉醇的化疗敏感性。

关键词: SIRT6, 骨肉瘤, siRNA

Abstract: Background and purpose: Sirtuin 6 (SIRT6) is an important histone modifying protein that is implicated in diverse physiological processes, as well as aging-associated diseases, and regulates DNA repair, energy metabolism and target gene expression. However, the role of SIRT6 in human osteosarcoma has not been fully understood. This study investigated the effect of the small interfering RNA (siRNA)-mediated SIRT6 knockdown on the survival and invasion. Methods: The siRNA against SIRT6 was constructed and transfected into U2OS cell line with Lipofectamine^TM 2000. The expression of SIRT6 was detected by Western blot. Flow cytometry was used to detect the cell apoptosis. Proliferation of cells was assayed using the cell counting kit-8 (CCK-8) method. The Transwell test was used to detect the invasion and migration ability of U2OS cells. And the viability in taxol was detected using CCK-8 method. Results: Compared with the control group, the protein level was significantly decreased in the experimental group at 24, 48 and 72 hours after transfection with SIRT6 siRNA. Flow cytometry detection showed the apoptotic rate increased significantly in the experimental group (P<0.05). Cell invasion and migration ability in the experimental group decreased significantly (P<0.05). In addition, the experimental group showed decreased growth after exposure to taxol compared with the control group (P<0.05). Conclusion: Down-regulation of SIRT6 expression may decrease invation and migration, promote apoptosis, and enhance the sensitivity to Taxol in U2OS osteosarcoma cell line.

Key words: SIRT6, Osteosarcoma, siRNA

余　天,李　娟,张伟凯,等. siRNA抑制骨肉瘤U2OS细胞系中SIRT6的表达及其生物学作用研究[J]. 中国癌症杂志, 2018, 28(10): 721-725.

YU Tian, LI Juan, ZHANG Weikai, et al. Influence of down-regulation of SIRT6 expression by small interfering RNA on U2OS osteosarcoma cell line[J]. China Oncology, 2018, 28(10): 721-725.

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siRNA抑制骨肉瘤U2OS细胞系中SIRT6的表达及其生物学作用研究

Influence of down-regulation of SIRT6 expression by small interfering RNA on U2OS osteosarcoma cell line

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