考利宁通过靶向 Pim-3 抑制食管鳞癌细胞生长。

Corynoline inhibits esophageal squamous cell carcinoma growth via targeting Pim-3.

机构信息

The Pathophysiology Department, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450000, China; China-US (Henan) Hormel Cancer Institute, Zhengzhou 450000, China; Tianjian Laboratory for Advanced Biomedical Sciences, Zhengzhou, Henan 450052, China.

The Pathophysiology Department, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450000, China.

出版信息

Phytomedicine. 2024 Jan;123:155235. doi: 10.1016/j.phymed.2023.155235. Epub 2023 Dec 12.

DOI:10.1016/j.phymed.2023.155235

PMID:38128397

Abstract

BACKGROUND

Esophageal squamous cell carcinoma (ESCC) is an aggressive and deadly malignancy characterized by late-stage diagnosis, therapy resistance, and a poor 5-year survival rate. Finding novel therapeutic targets and their inhibitors for ESCC prevention and therapy is urgently needed.

METHODS

We investigated the proviral integration site for maloney murine leukemia virus 3 (Pim-3) protein levels using immunohistochemistry. Using Methyl Thiazolyl Tetrazolium and clone formation assay, we verified the function of Pim-3 in cell proliferation. The binding and inhibition of Pim-3 by corynoline were verified by computer docking, pull-down assay, cellular thermal shift assay, and kinase assay. Cell proliferation, Western blot, and a patient-derived xenograft tumor model were performed to elucidate the mechanism of corynoline inhibiting ESCC growth.

RESULTS

Pim-3 was highly expressed in ESCC and played an oncogenic role. The augmentation of Pim-3 enhanced cell proliferation and tumor development by phosphorylating mitogen-activated protein kinase 1 (MAPK1) at T185 and Y187. The deletion of Pim-3 induced apoptosis with upregulated cleaved caspase-9 and lower Bcl2 associated agonist of cell death (BAD) phosphorylation at S112. Additionally, binding assays demonstrated corynoline directly bound with Pim-3, inhibiting its activity, and suppressing ESCC growth.

CONCLUSIONS

Our findings suggest that Pim-3 promotes ESCC progression. Corynoline inhibits ESCC progression through targeting Pim-3.

摘要

背景

食管鳞状细胞癌（ESCC）是一种侵袭性和致命性的恶性肿瘤，其特点是诊断晚期、治疗耐药和 5 年生存率低。因此，迫切需要寻找新的治疗靶点及其抑制剂，用于 ESCC 的预防和治疗。

方法

我们使用免疫组织化学方法研究了莫洛尼鼠白血病病毒 3（Pim-3）蛋白水平的前病毒整合位点。通过噻唑蓝（MTT）比色法和克隆形成实验，验证了 Pim-3 在细胞增殖中的功能。通过计算机对接、下拉实验、细胞热转移实验和激酶实验验证了 Corynoline 与 Pim-3 的结合和抑制作用。通过细胞增殖实验、Western blot 和患者来源的异种移植肿瘤模型，阐明了 Corynoline 抑制 ESCC 生长的作用机制。

结果

Pim-3 在 ESCC 中高表达，发挥致癌作用。Pim-3 的增强可通过磷酸化丝裂原活化蛋白激酶 1（MAPK1）的 T185 和 Y187 增强细胞增殖和肿瘤发展。Pim-3 的缺失诱导细胞凋亡，同时上调 cleaved caspase-9 并降低 Bcl2 相关细胞死亡激动剂（BAD）的 S112 磷酸化。此外，结合实验表明 Corynoline 可直接与 Pim-3 结合，抑制其活性，并抑制 ESCC 生长。