Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(4):1434-1442. doi: 10.26355/eurrev_201902_17100.
To discover the effect and mechanism of exogenous microRNA-29b (miR-29b) on proliferation, apoptosis and the sensitivity to chemotherapy of osteosarcoma (OS) cells.
We assessed the expression of microRNA-29b in osteosarcoma tissues evaluating the regulation of in on the OS cell growth and drug sensitivity in human osteosarcoma MG-63 cell model. Firstly, quantitative RT-PCR (reverse transcription-PCR, RT-PCR) was used to measure the expression of miR-29b and matrix metallopeptidase 9(MMP-9) in primary osteosarcoma samples, and to evaluate the correlation between the two molecules. Secondly, miR-29b mimics or mimics were used to modify its expression in MG-63 cells. Luciferase reporter assay, Western blotting, cell viability, colony forming assay and apoptosis examination were performed to assess the regulation by manipulated miR-29b in the osteosarcoma-derived cells.
We found that miR-29b is down-expressed, whereas the MMP-9 level was markedly higher in primary osteosarcoma tissues and osteosarcoma-derived cells. We also found that exogenous miR-29b reduces the proliferation, promotes the apoptosis and upregulates the sensitivity to chemotherapy (doxorubicin) of osteosarcoma cells via direct targeting of the MMP-9.
Our data suggest that the reduced miRNA-29b may serve as a predictor of response to chemotherapy and as a therapeutic target in human osteosarcomas.
探索外源性 microRNA-29b（miR-29b）对骨肉瘤（OS）细胞增殖、凋亡及化疗敏感性的影响及机制。
我们评估了骨肉瘤组织中 microRNA-29b 的表达，在人骨肉瘤 MG-63 细胞模型中评估了 miR-29b 对 OS 细胞生长和药物敏感性的调节作用。首先，采用反转录-PCR（RT-PCR）定量检测了骨肉瘤原代样本中 miR-29b 和基质金属蛋白酶 9（MMP-9）的表达，并评估了这两个分子之间的相关性。其次，采用 miR-29b 模拟物或模拟物修饰 MG-63 细胞中的表达。采用荧光素酶报告基因检测、Western blot、细胞活力检测、集落形成实验和凋亡检测，评估经人为调控 miR-29b 对骨肉瘤源性细胞的调节作用。
我们发现，miR-29b 在骨肉瘤原代组织和骨肉瘤源性细胞中呈低表达，而 MMP-9 水平明显升高。我们还发现，外源性 miR-29b 通过直接靶向 MMP-9，减少骨肉瘤细胞的增殖，促进其凋亡，并提高其对化疗药物（阿霉素）的敏感性。
我们的数据表明，miR-29b 的减少可能作为骨肉瘤患者对化疗反应的预测因子，并可作为治疗靶点。
