Section Pathology of The Institute of Forensic Medicine, University Hospital Jena, Friedrich Schiller University, D‑07747 Jena, Germany.
Leibniz Institute on Ageing‑Fritz Lipmann Institute (FLI), D‑07745 Jena, Germany.
Oncol Rep. 2020 Jun;43(6):2073-2082. doi: 10.3892/or.2020.7546. Epub 2020 Mar 17.
Mutations of p53 occur in approximately 50% of advanced non‑small cell lung cancer (NSCLC) cases, leading to loss of tumor suppressive function and/or gain of p53 oncogenic activity. Reactivation of mutant p53 and consequently induction of apoptosis in cancer cells is the goal of p53‑targeted therapy. Recently, several p53 mutant reactivating compounds were discovered including SCH 529074. However, the role of SCH 529074 in NSCLC has not been fully explored. In the present study, the effects of this compound on cell survival, cell cycle progression, induction of apoptosis and modulation of cell signaling in p53 mutant NSCLC cells (H1975, H322 and H157) and p53 wild‑type NSCLC cells (A549), was investigated. Cell‑based functional assays, real‑time RT‑qPCR and western blot assays were used. HCT116 [p53 wild‑type (WT)] and HCT116 p53‑/‑ (p53 null) were used as control cells. The results demonstrated that SCH 529074 treatment caused significant reduction in cell viability and colony formation activity in p53 mutant, p53 WT and p53‑deficient cells. The treatment of NSCLC cells with SCH 529074 resulted in a dose‑dependent induction of apoptosis and G0/G1 cell cycle arrest, which was associated with the activation of caspases (3 and 7), p53‑independent upregulation of p21 and PUMA as well as increased LC3II, a biomarker of autophagy. The combination treatment with the autophagy inhibitor chloroquine (CQ) and SCH 529074 led to decreased cell viability, colony formation and increased induction of apoptosis. The data indicated that SCH 529074 may exert its growth inhibitory function in a p53‑independent manner in NSCLC cells.
p53 突变发生于大约 50%的晚期非小细胞肺癌（NSCLC）病例中，导致肿瘤抑制功能丧失和/或获得 p53 致癌活性。p53 靶向治疗的目的是使突变型 p53 重新激活，从而诱导癌细胞凋亡。最近，发现了几种 p53 突变体再激活化合物，包括 SCH529074。然而，SCH529074 在 NSCLC 中的作用尚未完全探索。在本研究中，研究了该化合物对 p53 突变型 NSCLC 细胞（H1975、H322 和 H157）和 p53 野生型 NSCLC 细胞（A549）中的细胞存活、细胞周期进程、凋亡诱导和细胞信号转导的调节作用。使用基于细胞的功能测定、实时 RT-qPCR 和 Western blot 测定。使用 HCT116[p53 野生型（WT）]和 HCT116 p53-/-（p53 缺失）作为对照细胞。结果表明，SCH529074 处理导致 p53 突变、p53 WT 和 p53 缺陷细胞的细胞活力和集落形成活性显著降低。SCH529074 处理 NSCLC 细胞导致剂量依赖性的细胞凋亡和 G0/G1 细胞周期阻滞，这与半胱天冬酶（3 和 7）的激活、p53 非依赖性上调 p21 和 PUMA 以及增加自噬标志物 LC3II 有关。自噬抑制剂氯喹（CQ）与 SCH529074 的联合治疗导致细胞活力、集落形成降低和凋亡诱导增加。数据表明，SCH529074 可能以 p53 非依赖性方式在 NSCLC 细胞中发挥其生长抑制功能。
