摘要
目的研究全反式维甲酸对143B骨肉瘤细胞生长的抑制作用及可能的机制。方法利用RT-PCR(ReverseTranscription Polymerase Chain Reaction)和Western blot检测RAR和RXR在骨肉瘤细胞143B和MG63的内源性表达。通过细胞计数证实全反式维甲酸抑制143B细胞生长,利用凋亡染色和Western blot,检测全反式维甲酸是否诱导143B细胞凋亡;利用萤光素酶报告质粒测定成骨分化早期转录调节因子Runx2的转录活性,用Western blot检测骨桥素和骨钙素的表达,确证全反式维甲酸对143B细胞成骨分化不同时期标志物的影响。结果 RAR及RXR在骨肉瘤细胞143B和MG63中均存在内源性表达。全反式维甲酸能明显呈浓度依赖性抑制143B骨肉瘤细胞的生长;全反式维甲酸处理后,Caspase-3蛋白水平增加,且凋亡染色出现明显阳性;全反式维甲酸不仅使Runx2转录活性增加,而且也促进骨桥素和骨钙素的表达。结论全反式维甲酸能明显抑制143B骨肉瘤细胞生长,这种作用至少与其通过维甲酸信号通路促进143B细胞成骨分化有关。
Aim To study the inhibitory effect of all trans retinoic acid（ATRA） on 143B osteosarcoma cells and the possible mechanism.Methods First,the endogenous retinoids nuclear receptor expression in 143B and MG63 osteosarcoma cells were tested,Then cell counting was employed to test the proliferation inhibitory effect of ATRA on 143B cells.Western blot was used to test the caspase-3 level,as well as apoptosis staining,to make sure that ATAR can not only inhibits proliferation but also induces apoptosis in 143B cells.Finally,the luciferase reporter assay was adopt-ed to check the osteogenic transcription factor activity of Runx2,the protein expression of osteopontin and osteocalcin were measured with western blot.Results Different types of retinoids nuclear receptor were detected in 143B and MG63 osteosarcoma cells.ATRA inhibited the proliferation of 143B cells in a concentration-dependent manner,The caspase-3 increased apparently after the cells were treated with ATRA for 12 hour and the apoptosis staining was positive apparently.The transcription activity of Runx2 increased,so did the protein level of osteopontin and osteocalcin.Conclusion ATRA can inhibit the proliferation of 143B osteosarcoma cells and induce the cell apoptosis.These effects may at least result from the osteogenic differentiation of osteosarcoma cells initialized by ATRA via retinoic acid signaling.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第8期1091-1095,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No81071462)
关键词
全反式维甲酸
骨肉瘤
增殖抑制
凋亡
成骨分化
维甲酸信号
all trans retinoic acid
osteosarcoma
proliferation inhibiting
apoptosis
osteogenic differentiation
retinoic acid signaling
