Karaliotas Georgios I, Mavridis Konstantinos, Scorilas Andreas, Babis George C
First Orthopedic Department, Athens University Medical School, Attikon University Hospital, Haidari, Athens 12462, Greece.
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimiopolis, Athens 15701, Greece.
Mol Med Rep. 2015 Sep;12(3):4514-4521. doi: 10.3892/mmr.2015.3939. Epub 2015 Jun 16.
Osteoarthritis (OA) is primarily characterized by articular cartilage degeneration and chondrocyte loss. Although the role of apoptosis in cartilage pathobiology remains to be elucidated, the apoptotic B‑cell CLL/lymphoma 2 (BCL2) gene family is considered to be involved in OA. The purpose of the present study was to quantitatively analyze the mRNA expression profiles of the BCL2‑associated X protein (BAX) and BCL2 genes in human OA and in normal cartilage. Cartilage tissue samples were obtained from 78 patients undergoing total knee arthroplasty for OA (OA group) and orthopedic interventions for causes other than OA (control group). Total RNA was isolated from the cartilage tissue specimens and reverse transcribed into cDNA. A highly sensitive and specific reverse transcription quantitative polymerase chain reaction assay was developed for quantification of the mRNA levels of BAX and BCL2, using beta‑2 microglobulin as an endogenous control for normalization purposes. Gene expression analysis was performed using the comparative Ct (2(‑ΔΔCt)) method. The mRNA expression of BAX presented an increasing trend in the OA group compared with the control group, although without statistically significace (P=0.099). By contrast, the expression ratio of BCL2/BAX was found to be significantly decreased (2.76‑fold) in the OA group compared with the normal cartilage control group (P=0.022). A notable 4.6‑fold overexpression of median mRNA levels of BAX was also observed in patients with stage III OA compared with the control (P=0.034), while the BCL2/BAX ratio was markedly (2.5‑fold) decreased (P=0.024). A marked positive correlation was observed between the mRNA levels of BAX and BCL2 in the control group (r(s)=0.728; P<0.001), which was also present in the OA group, although to a lesser degree (r(s)=0.532; P<0.001). These results further implicate apoptosis in the pathogenesis of OA, through molecular mechanisms, which include the aberrant expression of the BCL2 gene family. Further investigation may reveal novel prognostic biomarkers and potential targets for therapeutic interventions in the early stages of OA.
骨关节炎（OA）的主要特征是关节软骨退变和软骨细胞丢失。尽管细胞凋亡在软骨病理生物学中的作用仍有待阐明，但凋亡相关的B细胞淋巴瘤2（BCL2）基因家族被认为与OA有关。本研究的目的是定量分析人OA软骨和正常软骨中B细胞淋巴瘤2相关X蛋白（BAX）和BCL2基因的mRNA表达谱。从78例因OA接受全膝关节置换术的患者（OA组）和因OA以外原因接受骨科干预的患者（对照组）获取软骨组织样本。从软骨组织标本中分离总RNA并逆转录成cDNA。建立了一种高灵敏度和特异性的逆转录定量聚合酶链反应检测方法，以β-2微球蛋白作为内参进行标准化，定量BAX和BCL2的mRNA水平。使用比较Ct（2（-ΔΔCt））方法进行基因表达分析。与对照组相比，OA组中BAX的mRNA表达呈上升趋势，尽管无统计学意义（P = 0.099）。相比之下，与正常软骨对照组相比，OA组中BCL2/BAX的表达比值显著降低（2.76倍）（P = 0.022）。与对照组相比，III期OA患者中BAX的中位mRNA水平也有显著的4.6倍过表达（P = 0.034），而BCL2/BAX比值显著降低（2.5倍）（P = 0.024）。对照组中BAX和BCL2的mRNA水平之间存在显著正相关（r(s)=0.728；P<0.001），OA组中也存在这种相关性，尽管程度较轻（r(s)=0.532；P<0.001）。这些结果通过包括BCL2基因家族异常表达在内的分子机制，进一步表明细胞凋亡参与了OA的发病机制。进一步的研究可能会揭示OA早期阶段新的预后生物标志物和潜在的治疗干预靶点。
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