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1.400010 重庆,重庆医科大学附属第二医院骨科
2.400016 重庆,重庆市生物化学与分子药理学重点实验室
3.400016 重庆,重庆医科大学药学院药理学教研室
[ 尹良军,E-mail:albertyin@163.com YIN Liangjun, E-mail:albertyin@163.com
收稿:2016-10-21,
修回:2016-12-07,
纸质出版:2017-07-15
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吴念, 张平, 田冬冬, 等. ATRA与BMP9对人骨肉瘤细胞增殖及分化作用的研究[J]. 第三军医大学学报, 2017,39(13):1333-1338.
Nian WU, Ping ZHANG, Dongdong TIAN, et al. Effects of all-trans-retinoic acid and bone morphogenetic protein 9 on proliferation and differentiation of human osteosarcoma cells[J]. Journal of Third Military Medical University, 2017, 39(13): 1333-1338.
吴念, 张平, 田冬冬, 等. ATRA与BMP9对人骨肉瘤细胞增殖及分化作用的研究[J]. 第三军医大学学报, 2017,39(13):1333-1338. DOI: 10.16016/j.1000-5404.201610139.
Nian WU, Ping ZHANG, Dongdong TIAN, et al. Effects of all-trans-retinoic acid and bone morphogenetic protein 9 on proliferation and differentiation of human osteosarcoma cells[J]. Journal of Third Military Medical University, 2017, 39(13): 1333-1338. DOI: 10.16016/j.1000-5404.201610139.
目的
2
研究全反式维甲酸(all-trans-retinoic acid,ATRA)与骨形态发生蛋白9(bone morphogenetic proteins 9,BMP9) 对人骨肉瘤细胞增殖及分化的影响。
方法
2
利用四甲基偶氮唑蓝(MTT)法和流式细胞仪分析细胞增殖情况;蛋白质免疫印迹实验(Western blot)检测细胞增殖和成骨相关指标;半定量PCR及Western blot分析ATRA和ATRA与BMP9联用处理143B细胞株后对BMP9表达水平的影响。
结果
2
在人骨肉瘤143B细胞株中,与对照组相比,ATRA能明显抑制细胞增殖(
P
<
0.05),并促使细胞周期阻滞在G
1
期(
P
<
0.05),下调PCNA的蛋白水平,并呈浓度依赖性;ATRA能呈浓度依赖性增加晚期成骨指标OCN和OPN的蛋白水平(
P
<
0.05);ATRA能上调BMP9的mRNA及蛋白水平,并呈浓度依赖性(
P
<
0.05);单用BMP9并不能上调OCN和OPN表达,并促进其增殖(
P
<
0.05),但与ATRA联用时,能够增强ATRA诱导的相关成骨指标表达和抗增殖作用(
P
<
0.05)。
结论
2
ATRA对143B细胞的增殖具有抑制作用并诱导其成骨分化,并可能恢复BMP9的成骨诱导能力。
Objective
2
To determine the effects of all-trans-retinoic acid (ATRA) and bone morphogenetic protein 9 (BMP9) on the proliferation and osteogenic differentiation of human osteosarcoma (OS) cells.
Methods
2
MTT assay and flow cytometry were used to analyze the proliferation of the human OS cell line 143B. Western blot analysis was used to detect the proliferation-and osteogenesis-related markers. Semi-quantitative PCR and Western blot analysis were used to analyze the expression levels of BMP9 in 143B cells after treated with ATRA alone or combined with BMP9.
Results
2
In the 143B cells
ATRA significantly inhibited the proliferation of 143B cells (
P
<
0.05) compared with the untreated group
promoted cell cycle arrested in G1 phase (
P
<
0.05)
and reduced the pro
tein level of PCNA in
a
concentration-dependent manner. The protein levels of lately osteogenic makers OPN and OCN were up-regulated by ATRA treatment in
a
concentration-dependent manner (
P
<
0.05)
so were the mRNA and protein levels of endogenous BMP9 (
P
<
0.05). BMP9 alone failed to up-regulate the expression of OPN and OCN but promoted the proliferation of 143B cells (
P
<
0.05)
but when combined with ATRA
BMP9 enhanced the expression of osteogenic markers and anti-proliferation effect induced by ATRA (
P
<
0.05).
Conclusion
2
ATRA inhibit the proliferation and induce osteogenic differentiation in 143B cells
and may restore the osteogenic capability of BMP9.
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