Lv Hong-Wei, Xing Wen-Qun, Ba Yu-Feng, Li Hao-Miao, Wang Hao-Ran, Li Yin
Department of Thoracic Surgery, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou 450008, Henan Province, People's Republic of China.
Department of Thoracic Surgery, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou 450008, Henan Province, People's Republic of China; Department of Thoracic Surgery, The Cancer Hospital Chinese Academy of Medical Science, Beijing 100021, People's Republic of China.
Transl Oncol. 2021 Jun;14(6):101075. doi: 10.1016/j.tranon.2021.101075. Epub 2021 Mar 25.
Cisplatin (DDP) remains the backbone of chemotherapy for non-small cell lung cancer (NSCLC), yet its clinical efficacy is limited by DDP resistance. We aim to investigate the role of the SET and MYND domain-containing protein 3 (SMYD3) in DDP resistance of NSCLC.
Expression pattern of SMYD3 was determined in NSCLC tissues using qRT-PCR, which also validated its correlation with NSCLC clinicopathological stages. Impacts of SMYD3 on DDP resistance were evaluated by knocking down SMYD3 in DDP-resistant cells and overexpressing it in DDP-sensitive cells, and assessed for several phenotypes: IC by MTT, long-term proliferation by colony formation, apoptosis and cell-cycle distribution by flow cytometry. The interaction between Ankyrin Repeat and KH Domain Containing 1 (ANKHD1) and SMYD3 was examined by co-immunoprecipitation and immunofluorescence. The transcriptional regulation of SMYD3 on cyclin-dependent kinase 2 (CDK2) promoter regions was confirmed using chromatin-immunoprecipitation. The in vivo experiments using DDP-resistant cells with altered SMYD3 and ANKHD1 expression were further performed to verify the SMYD3/ANKHD1 axis.
Highly expressed SMYD3 was observed in NSCLC tissues or cells, acted as a sensitive indicator for NSCLC, correlated with higher TNM stages or resistant to DDP treatment, and shorter overall survival. The promotion of SMYD3 on DDP resistance requires co-regulator, ANKHD1. CDK2 was identified as a downstream effector. In vivo, SMYD3 knockdown inhibited the growth of DDP-resistant NSCLC cells, which was abolished by ANKHD1 overexpression.
SMYD3 confers NSCLC cells chemoresistance to DDP in an ANKHD1-dependent manner, providing novel therapeutic targets to overcome DDP resistance in NSCLC .
顺铂（DDP）仍然是非小细胞肺癌（NSCLC）化疗的主要药物，但其临床疗效受到DDP耐药性的限制。我们旨在研究含 SET 和 MYND 结构域蛋白 3（SMYD3）在 NSCLC 的 DDP 耐药中的作用。
使用 qRT-PCR 检测 NSCLC 组织中 SMYD3 的表达模式，并验证其与 NSCLC 临床病理分期的相关性。通过在 DDP 耐药细胞中敲低 SMYD3 以及在 DDP 敏感细胞中过表达 SMYD3，评估 SMYD3 对 DDP 耐药性的影响，并检测几种表型：通过 MTT 法检测 IC，通过集落形成检测长期增殖，通过流式细胞术检测凋亡和细胞周期分布。通过免疫共沉淀和免疫荧光检测锚蛋白重复序列和 KH 结构域包含蛋白 1（ANKHD1）与 SMYD3 之间的相互作用。使用染色质免疫沉淀法确认 SMYD3 对细胞周期蛋白依赖性激酶 2（CDK2）启动子区域的转录调控。进一步进行体内实验，使用改变了 SMYD3 和 ANKHD1 表达的 DDP 耐药细胞，以验证 SMYD3/ANKHD1 轴。
在 NSCLC 组织或细胞中观察到 SMYD3 高表达，它是 NSCLC 的一个敏感指标，与较高的 TNM 分期相关或对 DDP 治疗耐药，且总生存期较短。SMYD3 对 DDP 耐药性的促进作用需要共调节因子 ANKHD1。CDK2 被确定为下游效应分子。在体内，敲低 SMYD3 抑制了 DDP 耐药 NSCLC 细胞的生长，而 ANKHD1 的过表达消除了这种抑制作用。
SMYD3 以 ANKHD1 依赖的方式赋予 NSCLC 细胞对 DDP 的化学抗性，为克服 NSCLC 中的 DDP 耐药性提供了新的治疗靶点。
