Department of Orthopedics Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing Medical University, No. 1 Youyi Road, Yuzhong, Chongqing, 400010, China.
Department of Plastic Surgery, Chongqing University Central Hospital, Chongqing University, No. 1 Jiangkang Road, Yuzhong, Chongqing, 400010, China.
Curr Cancer Drug Targets. 2023;23(6):496-504. doi: 10.2174/1568009623666230206154944.
Protein kinase, membrane-associated tyrosine/threonine 1 (PKMYT1) contributes to the proliferative, migratory, invasive and colony-forming capabilities of oncocytes. Dysregulated expression of PKMYT1 is associated with numerous malignancies. However, at present, the functional role of PKMYT1 in osteosarcoma is still not clarified.
The present study, therefore, aimed to investigate the prognostic value of PKMYT1 in osteosarcoma, and to explore the underlying molecular mechanism(s).
To meet this end, the expression level of PKMYT1 in osteosarcoma was measured by immunohistochemical analysis. The prognostic value of PKMYT1 in osteosarcoma was analyzed on the basis of R2: Genomics Analysis and Visualization Platform. The functional role of PKMYT1 was subsequently investigated in MG63 cells by knocking down PKMYT1 expression lentivirus encoding shRNA. MTT assay, scratch-wound and Transwell assays were then used to determine whether PKMYT1 fulfills a role in the proliferative and invasive capabilities of the MG63 cells. Subsequently, the role of PKMYT1 in the apoptosis of the cells was assessed using western blot and immunofluorescence analyses. Finally, to determine whether PKMYT1 exerts its role through the NF-κB pathway, fibroblast-stimulating lipopeptide-1 (FSL-1) was used as an NF-κB activator.
Compared with normal tissues, osteosarcoma tissues showed a significantly increased level of PKMYT1 expression. The clinical survival analysis indicated that patients with high PKMYT1 expression were associated with lower probabilities of overall survival and metastasis-free survival compared with those with low PKMYT1 expression levels. Knockdown of PKMYT1 inhibited the migratory and invasive capabilities of the MG63 cells, and also facilitated their apoptosis. Moreover, the knockdown of PKMYT1 restrained the NF-κB pathway in MG63 cells, whereas activating the NF- κB pathway ameliorated the effects of silencing PKMYT1 on MG63 cells, suggesting that PKMYT1 functions the NF-κB pathway in MG63 cells.
Taken together, the results of the present study have shown that a high expression level of PKMYT1 is associated with poor prognosis of osteosarcoma, and that PKMYT1 is able to aggravate the malignant progression of MG63 cells negatively regulating the NF-κB pathway, suggesting that PKMYT1 may be a potential molecular therapeutic target for the treatment of osteosarcoma.
蛋白激酶，膜相关酪氨酸/苏氨酸 1（PKMYT1）有助于成骨细胞瘤的增殖、迁移、侵袭和集落形成能力。PKMYT1 的失调表达与许多恶性肿瘤有关。然而，目前 PKMYT1 在骨肉瘤中的功能作用尚不清楚。
因此，本研究旨在探讨 PKMYT1 在骨肉瘤中的预后价值，并探讨其潜在的分子机制。
为此，通过免疫组织化学分析测量骨肉瘤中 PKMYT1 的表达水平。基于 R2：基因组分析和可视化平台分析 PKMYT1 在骨肉瘤中的预后价值。随后，通过慢病毒编码 shRNA 敲低 PKMYT1 表达，在 MG63 细胞中研究 PKMYT1 的功能作用。然后，通过 MTT 测定、划痕实验和 Transwell 测定来确定 PKMYT1 是否在 MG63 细胞的增殖和侵袭能力中发挥作用。随后，通过 Western blot 和免疫荧光分析评估 PKMYT1 在细胞凋亡中的作用。最后，为了确定 PKMYT1 是否通过 NF-κB 途径发挥作用，使用纤维母细胞刺激脂肽-1（FSL-1）作为 NF-κB 激活剂。
与正常组织相比，骨肉瘤组织中 PKMYT1 的表达水平显著增加。临床生存分析表明，与低 PKMYT1 表达水平的患者相比，高 PKMYT1 表达的患者总体生存率和无转移生存率较低。敲低 PKMYT1 抑制了 MG63 细胞的迁移和侵袭能力，同时促进了它们的凋亡。此外，在 MG63 细胞中敲低 PKMYT1 抑制了 NF-κB 途径，而激活 NF-κB 途径则改善了沉默 PKMYT1 对 MG63 细胞的作用，这表明 PKMYT1 在 MG63 细胞中通过 NF-κB 途径发挥作用。
综上所述，本研究结果表明，PKMYT1 高表达与骨肉瘤预后不良相关，PKMYT1 能够通过负调控 NF-κB 途径加重 MG63 细胞的恶性进展，提示 PKMYT1 可能是治疗骨肉瘤的潜在分子治疗靶点。
