Bone and Joint Research Laboratory, Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
J Orthop Surg Res. 2009 Dec 24;4:45. doi: 10.1186/1749-799X-4-45.
Matrix metalloproteinase (MMPs) synthesized and secreted from connective tissue cells have been thought to participate in degradation of the extracellular matrix. Increased MMPs activities that degrade proteoglycans have been measured in osteoarthritis cartilage. This study aims to suppress the expression of the MMP-3 gene in in vitro human chondrosarcoma using siRNA.
CELLS WERE CATEGORIZED INTO FOUR GROUPS: control (G.1); transfection solution treated (G.2); negative control siRNA treated (G.3); and MMP-3 siRNA treated (G.4). All four groups were further subdivided into two groups - treated and non-treated with IL-1beta- following culture for 48 and 72 h. We observed the effects of gene suppression according to cell morphology, glycosaminoglycan (GAG) and hyaluronan (HA) production, and gene expression by using real-time polymerase chain reaction (PCR).
In IL-1beta treated cells the apoptosis rate in G.4 was found to be lower than in all other groups, while viability and mitotic rate were higher than in all other groups (p < 0.05). The production of GAG and HA in G.4 was significantly higher than the control group (p < 0.05). MMP-3 gene expression was downregulated significantly (p < 0.05).
MMP-3 specific siRNA can inhibit the expression of MMP-3 in chondrosarcoma. This suggests that MMP-3 siRNA has the potential to be a useful preventive and therapeutic agent for osteoarthritis.
结缔组织细胞合成和分泌的基质金属蛋白酶（MMPs）被认为参与细胞外基质的降解。在骨关节炎软骨中，已测量到降解蛋白聚糖的 MMPs 活性增加。本研究旨在使用 siRNA 抑制体外人软骨肉瘤中 MMP-3 基因的表达。
将细胞分为四组：对照组（G.1）；转染溶液处理组（G.2）；阴性对照 siRNA 处理组（G.3）；和 MMP-3 siRNA 处理组（G.4）。所有四组进一步分为两组 - 在培养 48 和 72 小时后用 IL-1β处理和不处理。我们通过实时聚合酶链反应（PCR）观察根据细胞形态、糖胺聚糖（GAG）和透明质酸（HA）产生以及基因表达的基因抑制效果。
在 IL-1β处理的细胞中，G.4 中的细胞凋亡率低于其他所有组，而活力和有丝分裂率高于其他所有组（p < 0.05）。G.4 中的 GAG 和 HA 产生明显高于对照组（p < 0.05）。MMP-3 基因表达明显下调（p < 0.05）。
MMP-3 特异性 siRNA 可抑制软骨肉瘤中 MMP-3 的表达。这表明 MMP-3 siRNA 有可能成为骨关节炎的一种有用的预防和治疗剂。
