Department of Radiation and Medical Oncology, Hubei Key Laboratory of Tumor Biological Behaviors, Hubei Cancer Clinical Study Center, Zhongnan Hospital of Wuhan University, Wuhan 430071, China.
Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang 441000, China.
Biosci Rep. 2022 Oct 28;42(10). doi: 10.1042/BSR20221128.
The metallothionein 1 (MT1) family was previously shown to be involved in metal ion homeostasis, DNA damage, oxidative stress, and carcinogenesis. Our team's previous study showed that MT1X is most closely associated with ccRCC. However, its role in clear cell RCC (ccRCC) remains unclear. The present study aimed to demonstrate MT1X's prognostic value, potential biologic function, impact on the immune system, and influence on cell growth, the cell cycle, apoptosis, and migration in the setting of ccRCC. The relationship between clinical pathologic features and MT1X was analyzed using bioinformatics. We knocked down MT1X in the ccRCC cell line 786O with si-MT1X to verify the results of the bioinformatic analysis at the cytological level. Apoptosis assay, cell cycle assay, wound-healing assay, colony formation assay, and RT-qPCR were performed. MT1X is correlated with the stage (T and M) and grade and is able to be an independent prognostic factor for ccRCC. The TISIDB database analysis showed a significant correlation between MT1X and tumor-infiltrating lymphocytes such as central memory CD8+ T cells and γΔT cells. MT1X was also positively related to immunomodulators such as TGFB1 and CXCR4. We also found that MT1X knockdown inhibits cell growth, induces apoptosis, arrests cells in the S cell cycle, and inhibits the wound healing proportion in ccRCC. Gene set enrichment analysis and quantitative PCR (q-PCR) analysis found that down-regulation of MT1X reduced the accumulation of hypoxia-associated factors. Bioinformatic analysis associated increased MT1X expression with a worse prognosis. Laboratory experiments confirmed bioinformatic findings. MT1X was also found to be an independent prognostic biomarker for ccRCC and is involved in immune system regulation.
金属硫蛋白 1(MT1)家族先前被证明参与金属离子稳态、DNA 损伤、氧化应激和致癌作用。我们团队的先前研究表明 MT1X 与 ccRCC 最密切相关。然而,其在透明细胞肾细胞癌(ccRCC)中的作用仍不清楚。本研究旨在证明 MT1X 在 ccRCC 中的预后价值、潜在生物学功能、对免疫系统的影响以及对细胞生长、细胞周期、凋亡和迁移的影响。使用生物信息学分析分析临床病理特征与 MT1X 的关系。我们使用 si-MT1X 敲低 ccRCC 细胞系 786O 中的 MT1X,以在细胞学水平上验证生物信息学分析的结果。进行了凋亡测定、细胞周期测定、划痕愈合测定、集落形成测定和 RT-qPCR。MT1X 与分期(T 和 M)和分级相关,并且能够成为 ccRCC 的独立预后因素。TISIDB 数据库分析显示 MT1X 与中央记忆 CD8+T 细胞和γΔT 细胞等肿瘤浸润淋巴细胞以及 TGFB1 和 CXCR4 等免疫调节剂呈显著相关性。我们还发现 MT1X 敲低抑制 ccRCC 中的细胞生长、诱导凋亡、使细胞在 S 细胞周期中停滞并抑制伤口愈合比例。基因集富集分析和定量 PCR(q-PCR)分析发现,下调 MT1X 减少了与缺氧相关的因子的积累。生物信息学分析将 MT1X 表达增加与预后不良相关联。实验室实验证实了生物信息学发现。MT1X 也是 ccRCC 的独立预后生物标志物,并且参与免疫系统调节。