Analysis of human osteoarthritic connective tissue by laser capture microdissection and QRT-PCR.
Analysis of human osteoarthritic connective tissue by laser capture microdissection and QRT-PCR.
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通过激光捕获显微切割和 QRT-PCR 分析人类骨关节炎结缔组织。
DOI：
10.1080/03008200701692685
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发表时间：
2007
影响因子：
2.9
通讯作者：
Landis,WilliamJ
中科院分区：
文献类型：
--
作者：
Scharschmidt,Thomas;Jacquet,Robin;Laskovski,Jovan;Lowder,Elizabeth;Weiner,Scott;Landis,WilliamJ
II 型胶原蛋白和聚集蛋白聚糖的基因表达水平已被确定为接受全膝关节置换术的患者中人类骨关节炎的潜在指标和疾病标志物。在这方面，手术时获得的受影响关节软骨标本被置于 RNAlaterTM 中以保持 RNA 完整性，随后进行冷冻切片。通过激光捕获显微切割分离单个或少量软骨细胞，并通过定量逆转录聚合酶链反应提取和分析其总RNA。结果表明，使用这些方法可以检测和重现骨关节炎组织中特定细胞的 II 型胶原蛋白和聚集蛋白聚糖 mRNA 表达。我们的工作首次证明成功分离仅限于包含少量人类骨关节炎材料的软骨细胞的RNA。该研究提出了一种可以严格分析该疾病及其进展的新途径。
Gene expression levels for type II collagen and aggrecan have been determined as potential measures and disease markers of human osteoarthritis in patients undergoing total knee arthroplasty. In this regard, specimens of affected articular cartilage obtained intraoperatively at the time of surgery were placed in RNAlaterTMto maintain RNA integrity and subsequently frozen-sectioned. Individual or small numbers of chondrocytes were isolated by laser capture microdissection and their total RNA was extracted and analyzed by quantitative reverse transcription-polymerase chain reaction. Results indicate that type II collagen and aggrecan mRNA expression from specific cells in osteoarthritic tissues are detectable and reproducible using these approaches. Our work is the first to demonstrate successful isolation of RNA limited to chondrocytes comprising small quantities of human osteoarthritic material. The study presents a new avenue by which the disease and its progression may be critically assayed.