Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Hepato‑Pancreato‑Biliary Surgery, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.
Department of Breast Oncology, Tianjin Medical University Cancer Institute and Hospital, Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, P.R. China.
Oncol Rep. 2020 Aug;44(2):565-576. doi: 10.3892/or.2020.7635. Epub 2020 Jun 5.
Retroperitoneal liposarcoma (RLPS) is one of the most common types of retroperitoneal sarcomas, and has a high recurrence rate. There is an urgent need to further explore its pathogenesis and develop more effective treatment strategies. The aim of the present study was to identify potential driver genes of RLPS through bioinformatics analysis and molecular biology to elucidate potential targets that are suitable for further analysis for the treatment of RLPS. Differentially expressed genes (DEGs) between liposarcoma and normal fatty (NF) tissues were identified based on microarray data through bioinformatics analysis, and thymidylate synthase (TYMS) was selected from the DEGs, based on high content screening (HCS). TYMS expression was evaluated in RLPS tumor tissues and cell lines. A total of 21 RLPS tissues and 10 NF frozen tissues were used for reverse transcription‑quantitative PCR, and 47 RLPS formalin‑fixed specimens were used for immunohistochemical analysis. The effect of TYMS downregulation on cell proliferation, apoptosis, cell cycle progression, and cell migration and invasion were evaluated using lentivirus‑mediated short hairpin RNA. The underlying mechanisms of TYMS in RLPS were examined by protein microarray and verified by western blotting. A total of 855 DEGs were identified. TYMS knockdown had the most notable effect on the proliferative capacity of RLPS cells according to the HCS results. TYMS mRNA expression levels were higher in RLPS tissues compared with NF tissues (P<0.001). TYMS expression was higher in high‑grade RLPS tissues compared with low‑grade RLPS tissues (P=0.003). The patients with positive TYMS expression had a worse overall survival (OS) and disease‑free survival (DFS) compared with the patients with negative TYMS expression (OS, P=0.024; DFS, P=0.030). The knockdown of TYMS reduced proliferation, promoted apoptosis, facilitated cell cycle progression from G1 to S phase, and reduced cell migration and invasion of RLPS cells. Protein microarray analysis and western blotting showed that the Janus Kinase/Signal transducers and activators of transcription pathway was downregulated following TYMS knockdown. In conclusion, TYMS expression is upregulated in RLPS tissues, and downregulation of TYMS reduces RLPS progression.
腹膜后脂肪肉瘤 (RLPS) 是最常见的腹膜后肉瘤类型之一，具有很高的复发率。因此迫切需要进一步探讨其发病机制，并开发更有效的治疗策略。本研究旨在通过生物信息学分析和分子生物学鉴定 RLPS 的潜在驱动基因，以确定适合进一步分析用于治疗 RLPS 的潜在靶点。通过生物信息学分析，从基因芯片数据中鉴定出脂肪肉瘤与正常脂肪 (NF) 组织之间的差异表达基因 (DEGs)，并根据高内涵筛选 (HCS) 从 DEGs 中选择胸苷酸合成酶 (TYMS)。评估 RLPS 肿瘤组织和细胞系中 TYMS 的表达。共使用逆转录-定量 PCR 评估了 21 例 RLPS 组织和 10 例 NF 冷冻组织，并用免疫组织化学分析了 47 例 RLPS 福尔马林固定标本。使用慢病毒介导的短发夹 RNA 下调 TYMS 表达，评估对细胞增殖、凋亡、细胞周期进程以及细胞迁移和侵袭的影响。通过蛋白质微阵列检测 TYMS 在 RLPS 中的作用机制，并通过 Western blot 进行验证。鉴定出总共 855 个 DEGs。根据 HCS 结果，TYMS 敲低对 RLPS 细胞的增殖能力影响最显著。与 NF 组织相比，RLPS 组织中 TYMS mRNA 表达水平更高(P<0.001)。高级别 RLPS 组织中 TYMS 表达高于低级别 RLPS 组织(P=0.003)。TYMS 表达阳性的患者总生存期 (OS) 和无病生存期 (DFS) 较 TYMS 表达阴性的患者差(OS，P=0.024；DFS，P=0.030)。TYMS 敲低降低了 RLPS 细胞的增殖，促进了凋亡，促进了细胞周期从 G1 期向 S 期的进展，并减少了 RLPS 细胞的迁移和侵袭。蛋白质微阵列分析和 Western blot 显示，TYMS 敲低后 Janus 激酶/信号转导和转录激活物 (JAK/STAT) 通路被下调。综上所述，TYMS 在 RLPS 组织中表达上调，下调 TYMS 可降低 RLPS 的进展。
