[1]孙光雨),李 昊),裴艳涛).靶向SOX4基因的慢病毒感染对骨肉瘤MG63细胞凋亡的影响[J].郑州大学学报(医学版),2018,(06):759-763.[doi:10.13705/j.issn.1671-6825.2018.04.115]
 SUN Guangyu),LI Hao),PEI Yantao).Effects of lentiviral infection targeting SOX4 gene on apoptosis in osteosarcoma cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2018,(06):759-763.[doi:10.13705/j.issn.1671-6825.2018.04.115]
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靶向SOX4基因的慢病毒感染对骨肉瘤MG63细胞凋亡的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2018年06期
页码:
759-763
栏目:
应用研究
出版日期:
2018-11-20

文章信息/Info

Title:
Effects of lentiviral infection targeting SOX4 gene on apoptosis in osteosarcoma cells
作者:
孙光雨1)李 昊2)裴艳涛3)
1)肥城矿业中心医院骨一科 山东肥城 271608;2)山东大学齐鲁医院脊柱外科 济南 250000;3)山东大学齐鲁医院手足外科 济南250000
Author(s):
SUN Guangyu1)LI Hao2)PEI Yantao3)
1)Department of Orthopaedics,Feicheng Mining Center Hospital,Feicheng,Shandong 271608;2)Department of Spinal Surgery,Qilu Hospital of Shandong University,Jinan 250000;3)Department of Hand and Foot Surgery,Qilu Hospital of Shandong University,Jinan 250000
关键词:
骨肉瘤 SOX4基因 凋亡 Caspase-3 MG63细胞
Keywords:
osteosarcoma SOX4 gene apoptosis Caspase-3 MG63 cell
分类号:
R738.1
DOI:
10.13705/j.issn.1671-6825.2018.04.115
摘要:
目的:探讨靶向SOX4基因的慢病毒感染对骨肉瘤细胞凋亡的影响。方法:体外培养人成骨细胞株hFOB1.19和骨肉瘤细胞株MG63,采用qRT-PCR和Western blot检测两种细胞中SOX4 mRNA和蛋白的表达。将MG63细胞分为对照组、阴性组(感染LV-NC-GFP-shRNA)、干扰组(感染LV-SOX4-GFP-shRNA)和干扰+抑制剂组(LV-SOX4-GFP-shRNA感染前加Caspase-3抑制剂Ac-DEVD-CHO),采用qRT-PCR和Western blot检测对照组、阴性组和干扰组MG63细胞SOX4 mRNA和蛋白的表达情况,用于判断慢病毒感染效果; 采用流式细胞仪检测4组细胞凋亡,Western blot检测酶切Caspase-3蛋白的表达,Caspase-3活性检测试剂盒检测Caspase-3活性,采用JC-1染色检测线粒体膜电位。结果:与hFOB1.19细胞相比,MG63细胞中SOX4 mRNA和蛋白的表达水平升高(P<0.05); 与对照组相比,干扰组MG63细胞中SOX4 mRNA和蛋白的表达降低。干扰组较对照组细胞凋亡增加、Caspase-3活性和酶切蛋白的表达增强,线粒体膜电位降低(P均<0.05),而干扰+抑制剂组上述变化减弱(P<0.05)。结论:SOX4促进MG63细胞凋亡的作用机制可能与Caspase-3介导的凋亡途径有关。
Abstract:
Aim:To explore the effects of lentivirus infection targeting SOX4 gene on osteosarcoma cell apoptosis.Methods:Human osteoblast cell line hFOB1.19 and osteosarcoma cell line MG63 were cultured in vitro.The expressions of SOX4 mRNA and protein were detected by qRT-PCR and Western blot.The MG63 cells were divided into control group(not treated), negative group(negative control LV-NC-GFP-shRNA plasmid infection), interference group(LV-SOX4-GFP-shRNA plasmid infection)and interference+inhibitor group(Caspase-3 inhibitor Ac-DEVD-CHO treatment was added before LV-SOX4-GFP-shRNA plasmid infection). The expressions of SOX4 mRNA and protein in the first 3 groups were detected by qRT-PCR and Western blot to assess the effect of lentivirus infection.The rate of cell apoptosis was detected by flow cytometry,the expression of Caspase-3 protein was tested by Western blot,Caspase-3 activity was measured by Caspase-3 activity assay kit,and the changes of mitochondrial membrane potential were detected by JC-1 staining.Results:The expression levels of SOX4 mRNA and protein in MG63 cells were significantly higher than those of hFOB1.19 cells(P<0.05). Compared with control group,the expressions of SOX4 mRNA and protein in the interference group were reduced(P<0.05).The apoptosis rate of the cells in the interference group was increased,the expression levels of Caspase-3 activity and protein were enhanced,while MTP was decreased(P<0.05)compared with control group; after the addition of Caspase-3 inhibitor, the above changes were significantly weakened(P<0.05).Conclusion:Lentivirus infection targeting SOX4 gene can promote the apoptosis of MG63 cells, and its mechanism may be related to Caspase-3 mediated apoptosis pathway.

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备注/Memo

备注/Memo:
【基金项目】山东省自然科学基金资助项目(ZR2015HL096)
【作者简介】孙光雨,通信作者,男,1984年12月生,本科,主治医师,研究方向:关节、创伤及骨肿瘤,E-mail:ev3711@163.com
更新日期/Last Update: 2018-11-20