摘要
前期经基因芯片筛选发现,同源盒基因1(distal-less homeobox 1,DLX1)低表达是导致骨肉瘤形成的关键基因。该研究在此基础上,拟通过过表达DLX1探讨其对骨肉瘤细胞MG63迁移和侵袭等生物学特征的影响,并初步揭示其作用途径。采用含DLX1基因的重组腺病毒Ad DLX1(adenovirus distal-less homeobox 1)和空载腺病毒Ad RFP(adenovirus red fl uorescence protein)分别感染人骨肉瘤细胞MG63,观察细胞荧光表达情况,并用RT-PCR和Western blot验证DLX1表达水平;Transwell实验检测细胞迁移和侵袭能力;DAPI染色和流式细胞术检测细胞凋亡水平;CCK-8检测细胞增殖能力;RT-PCR和Western blot检测Wnt/β-catenin信号通路中β-catenin的表达。结果显示,与对照组(Ad RFP感染组)相比,Ad DLX1能显著增加MG63细胞中DLX1的表达,并导致细胞迁移和侵袭能力下降,但细胞增殖和凋亡情况无明显改变。DLX1过表达还可以上调Wnt/β-catenin信号通路中β-catenin的表达。该研究表明,DLX1可通过Wnt/β-catenin信号通路抑制骨肉瘤细胞MG63的迁移和侵袭。
To investigate the effects of overexpression of DLX1（distal-less homeobox 1） on migration, invasion, apoptosis and proliferation of osteosarcoma cell MG63, we infected the MG63 cells with Ad DLX1（adenovirus distal-less homeobox 1） and Ad RFP（adenovirus red fluorescence protein）. Then the expression level of DLX1 was confirmed by RT-PCR and Western blot. The cell migration and invasion were analyzed by Transwell assay. DAPI staining and flow cytometry were used to detect the cell apoptosis. The cell proliferation was detected by CCK-8 assay. The expression of β-catenin, a crucial molecule in Wnt/β-catenin pathway, was determined by RT-PCR and Western blot. The results showed that the m RNA and protein levels of DLX1 were siginificantly increased after infection with Ad DLX1, and the capacities of cell migration and invasion were down-regulated. Overexpression of DLX1 could not change the apoptosis and proliferation of MG63 cells. The expression of β-catenin m RNA and protein levels were up-regulated in MG63 cells. These results suggested that the Wnt/β-catenin pathway which involved in the migration and invasion of MG63 cells was inhibited by DLX1.
作者
吕凤香
康权
仇超
董姿杏
罗庆
Lu Fengxiang Kang Quan Qiu Chao Dong Zixing Luo Qing(Department of Pediatric Research Institution, Children＇s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of pediatrics, Chongqing 400014, China Department of Hepatology, Children＇s Hospital of Chongqing Medical University, Chongqing 400014, China)
出处
《中国细胞生物学学报》
CAS
CSCD
2017年第9期1165-1172,共8页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:81172545)
重庆市卫计委项目(批准号:2016MSXM039)资助的课题~~
