Department of Pathology, The First Affiliated Hospital of Sun Yat‑Sen University, Guangzhou, Guangdong 510080, P.R. China.
Mol Med Rep. 2018 Apr;17(4):5754-5763. doi: 10.3892/mmr.2018.8611. Epub 2018 Feb 15.
To investigate the roles of B‑cell lymphoma‑2 associated athanogene 3 (BAG3) in human chondrosarcoma and the potential mechanisms, the expression levels of BAG3 were detected in the present study, and the associations between BAG3 and clinical pathological parameters, clinical stage as well as the survival of patients were analyzed. The present study detected BAG3 mRNA and protein expression in the normal cartilage cell line HC‑a and in SW1353 chondrosarcoma cells by reverse transcription‑quantitative polymerase chain reaction and western blot analysis. The BAG3 protein expression in 59 cases of chondrosarcoma, 30 patients with endogenous chondroma and 8 cases of normal cartilage was semi-quantitatively analyzed using the immunohistochemical method. In addition, the BAG3 protein expression level, the clinical pathological parameters, clinical stage and the survival time of patients with chondrosarcoma were analyzed. The plasmid transfection method was employed to upregulate the expression BAG3 and small RNA interference to downregulate the expression of BAG3 in SW1353 cells. The expression levels of BAG3 protein and mRNA were significantly increased in the chondrosarcoma cell line when compared with the normal cartilage cell line. The immunohistochemistry results indicated that BAG3 protein was overexpressed in the tissue of human chondrosarcoma. Statistical analysis showed that the expression level of BAG3 was significantly increased in the different Enneking staging of patients with chondrosarcoma and Tumor staging, and there were no statistical differences in age, gender, histological classification and tumor size. In the in vitro experiments, the data revealed that BAG3 significantly promoted chondrosarcoma cell proliferation, colony‑formation, migration and invasion; however, it inhibited chondrosarcoma cell apoptosis. It was observed that BAG3 upregulated β‑catenin expression at the mRNA and protein levels. In addition, BAG3 induced the expression of runt‑related transcription factor 2 (RUNX2) in chondrosarcoma cells by upregulating β‑catenin. These clinical analyses revealed a positive association between β‑catenin and BAG3 in chondrosarcoma tumors. BAG3 was significantly increased in chondrosarcoma cells and tissues compared with the normal cartilage cells, tissue and cartilage benign tumors. Thus, BAG3 may serve as an oncogene in the development of chondrosarcoma via the induction of RUNX2 expression. The results of the present study contribute to further research on the biological development of chondrosarcoma.
为了探究 B 细胞淋巴瘤-2 相关抗凋亡基因 3(BAG3)在人软骨肉瘤中的作用及其潜在机制,本研究检测了 BAG3 的表达水平,并分析了 BAG3 与临床病理参数、临床分期以及患者生存之间的相关性。本研究通过逆转录-定量聚合酶链反应和 Western blot 分析检测了正常软骨细胞系 HC-a 和 SW1353 软骨肉瘤细胞中的 BAG3mRNA 和蛋白表达。采用免疫组织化学法半定量分析 59 例软骨肉瘤、30 例内生软骨瘤和 8 例正常软骨组织中 BAG3 蛋白的表达。此外,分析了软骨肉瘤患者的 BAG3 蛋白表达水平、临床病理参数、临床分期和生存时间。采用质粒转染法上调 BAG3 的表达,采用小干扰 RNA 下调 SW1353 细胞中 BAG3 的表达。与正常软骨细胞系相比,软骨肉瘤细胞系中 BAG3 蛋白和 mRNA 的表达水平显著升高。免疫组织化学结果表明,BAG3 蛋白在人软骨肉瘤组织中过表达。统计学分析显示,BAG3 的表达水平在软骨肉瘤患者的不同 Enneking 分期和肿瘤分期中显著增加,而在年龄、性别、组织学分类和肿瘤大小方面无统计学差异。在体外实验中,数据显示 BAG3 显著促进软骨肉瘤细胞的增殖、集落形成、迁移和侵袭,而抑制软骨肉瘤细胞的凋亡。结果表明,BAG3 可在 mRNA 和蛋白水平上上调 β-连环蛋白的表达。此外,BAG3 通过上调 β-连环蛋白诱导软骨肉瘤细胞中 runt 相关转录因子 2(RUNX2)的表达。这些临床分析显示,β-连环蛋白和 BAG3 在软骨肉瘤肿瘤中呈正相关。与正常软骨细胞、组织和软骨良性肿瘤相比,BAG3 在软骨肉瘤细胞和组织中明显增加。因此,BAG3 可能通过诱导 RUNX2 表达成为软骨肉瘤发生发展中的癌基因。本研究结果有助于进一步研究软骨肉瘤的生物学发展。