Departments of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia.
Departments of Biochemistry and Medicine, Virginia Commonwealth University, Richmond, Virginia.
J Cell Physiol. 2020 Oct;235(10):6862-6874. doi: 10.1002/jcp.29580. Epub 2020 Jan 27.
We have extended our analyses of (curcumin+sildenafil) biology. The drug combination caused vascularization and degradation of mutant K-RAS that correlated with reduced phosphorylation of ERK1/2, AKT T308, mTORC1, mTORC2, ULK1 S757, STAT3, STAT5, and NFκB and increased phosphorylation of eIF2α, ATM, AMPKα, ULK1 S317; all concomitant with elevated ATG13 S318 phosphorylation and autophagosome formation. Prior studies with drug combinations utilizing sildenafil have delineated an ATM-AMPK-ULK1 S317 pathway and an AKT-mTOR-ULK1 S757 pathway as modules which control ATG S318 phosphorylation and autophagosome formation. The knockdown of PKG reduced cell killing as well as reducing drug-enhanced phosphorylation of ATM, AMPKα, and ATG13. In the absence of PKG, no significant increase in ULK1 S317 phosphorylation was observed. In a Beclin1-dependent fashion, the drug combination reduced the expression of multiple histone deacetylase (HDAC) proteins, including HDAC2 and HDAC3. Molecular knockdown of HDAC2, HDAC3, and especially (HDAC2+HDAC3) significantly reduced the expression of PD-L1 and elevated expression of Class I human major histocompatibility complex. In vivo, (curcumin+sildenafil) enhanced the efficacy of 5-flurouracil against CT26 colorectal tumors. Prior exposure of established CT26 tumors to (curcumin+sildenafil) significantly enhanced the efficacy of a subsequently administered anti-PD-1 antibody. Collectively our data argue that (curcumin+sildenafil) has the potential in several settings to be an efficacious neoadjuvant therapy for colon cancer.
我们扩展了对（姜黄素+西地那非）生物学的分析。该药物组合导致血管生成和突变 K-RAS 的降解，这与 ERK1/2、AKT T308、mTORC1、mTORC2、ULK1 S757、STAT3、STAT5 和 NFκB 的磷酸化减少以及 eIF2α、ATM、AMPKα、ULK1 S317 的磷酸化增加有关；所有这些都伴随着 ATG13 S318 磷酸化和自噬体形成的增加。先前利用西地那非进行药物组合的研究已经描绘了 ATM-AMPK-ULK1 S317 途径和 AKT-mTOR-ULK1 S757 途径作为控制 ATG S318 磷酸化和自噬体形成的模块。PKG 的敲低降低了细胞杀伤作用，并降低了药物增强的 ATM、AMPKα 和 ATG13 的磷酸化。在没有 PKG 的情况下，没有观察到 ULK1 S317 磷酸化的显著增加。以 Beclin1 依赖的方式，药物组合降低了多种组蛋白去乙酰化酶（HDAC）蛋白的表达，包括 HDAC2 和 HDAC3。HDAC2、HDAC3 的分子敲低，特别是（HDAC2+HDAC3）显著降低了 PD-L1 的表达，并提高了 I 类人主要组织相容性复合物的表达。在体内，（姜黄素+西地那非）增强了 5-氟尿嘧啶对 CT26 结直肠肿瘤的疗效。预先暴露于 CT26 肿瘤的（姜黄素+西地那非）显著增强了随后给予的抗 PD-1 抗体的疗效。总的来说，我们的数据表明，（姜黄素+西地那非）在几种情况下有可能成为结直肠癌有效的新辅助治疗方法。
