DLX1联合BMP9促进人骨肉瘤细胞向成骨分化  

Effects of combination of DLX1 and BMP9 on inducing osteogeninc differentiation of human osteosarcoma cells

作　　者：吕凤香[1] 康权[1] 仇超[1] 董姿杏 罗庆[1] 

机构地区：[1]重庆医科大学附属儿童医院儿科研究所儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿科学重庆市重点实验室,重庆400014

出　　处：《基础医学与临床》2017年第6期768-774,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81172545);重庆市卫生和计划生育委员会医学科研项目(2016MSXM039)

摘　　要：目的探讨DLX1联合BMP9可否影响人骨肉瘤细胞MG63的成骨分化。方法用构建有DLX1和BMP9基因的重组腺病毒Ad DLX1和Ad BMP9,单独或联合感染MG63细胞,分为DLX1组(Ad DLX1+AdRFP感染组)、DLX1+BMP9组(Ad DLX1+Ad BMP9感染组)、BMP9组(Ad BMP9+AdRFP感染组)、RFP组(AdRFP感染组)。用RT-PCR和Western blot验证DLX1和BMP9的表达情况,Transwell实验检测细胞迁移、侵袭能力;碱性磷酸酶(ALP)染色和读数分析细胞早期成骨能力,茜素红染色检测细胞晚期成骨能力,Western blot检测骨桥蛋白(OPN)蛋白表达水平。结果 Ad DLX1和Ad BMP9感染MG63细胞后,DLX1和BMP9的表达水平上调(P<0.05);与RFP组相比,DLX1组、DLX1+BMP9组和BMP9组细胞迁移能力和侵袭能力下降(P<0.05),DLX1+BMP9组细胞迁移能力和侵袭能力下降更为明显(P<0.05);ALP活性、钙盐沉积和OPN蛋白表达在DLX1+BMP9组和BMP9组增加(P<0.05),且DLX1+BMP9组较BMP9组显著增强(P<0.05)。结论 DLX1与BMP9联合作用可抑制骨肉瘤细胞迁移和侵袭,同时可诱导骨肉瘤细胞向成骨分化。Objective To investigate the effects of Distal-less homeobox 1 （ DLX1 ） and bone morphogenetic protein 9 （BMP9） on inducing osteogeninc differentiation of human osteosarcoma cells. Methods The MC,63 cells were infected with AdDLX1 and AdBMP9 alone or in combination, grouped as DLX1 group（AdDLX1 ＋ AdRFP）, DLX1 ＋ BMP9 group （ AdDLX1 ＋ AdBMPg）, BMP9 group （ AdBMP9 ＋ AdRFP）, RFP group （AdRFP）. The expression of DLX1 and BMP9 were detected by RT-PCR and Western blot. The cell migration and invasion were analyzed by Transwell assay. Alkaline phosphatase（ALP） staining and numeration were used to detect the early osteogenic potential of MC,63 ceils, and the late osteogenic potential was detected by the alizarin red staining. The expression of osteopontin （OPN） protein was determined by Werstern blot assay. Results The mRNA and protein expression of DLX1 and BMP9 were increased by AdDLX1 and AdBMP9 （P 〈 0. 05 ）. Transwell assay resμLts showed the cell migration and invasion of DLX1 group, DLX1 ＋ BMP9 group and BMP9 group were decreased, DLX1 ＋ BMP9 group was the most one （P 〈 0. 05 ）. The activity of ALP and calcium deposition as well as OPN protein were increased in DLX1 ＋ BMP9 group and BMP9 group, these markers in DLX1 ＋ BMP9 group were more obvious（P 〈 O. 05）. Conclusions The combination of DLX1 and BMP9 may inhibit the malignant biological behavior and induce osteogenic differentiation of osteosarcoma cells.

关 键 词：骨肉瘤 DLX1 BMP9 恶性生物学行为 成骨分化 

分 类 号：R738.1[医药卫生—肿瘤]