Lv Jia-Jia, Song Wen-Ting, Li Xin-Min, Gao Jian-Mei, Yuan Ze-Li
Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi, China.
School of Pharmacy, Zunyi Medical University, Zunyi, China.
Front Pharmacol. 2020 Jun 26;11:911. doi: 10.3389/fphar.2020.00911. eCollection 2020.
Quinazoline derivatives display multiple pharmacological activities and target various biological receptors. Based on the skeleton of quinazoline core, we designed and synthesized three new quinazoline-phenyl chlormethine conjugates (I-III) bearing a Schiff base (C = N) linker, and investigated their anti-tumor effects on HepG2-xenografted tumor and human cancer cell line HepG2. Among these compounds, compound II showed better inhibitory effect against HepG2 cells. In the present study, TUNEL staining, western blot, molecular docking, and siRNA were used to investigate the inhibitory mechanism of compound II towards hepatoma. Compound II inhibited HepG2-xenografted tumor growth in nude mice. Moreover, Compound II not only up-regulated Bax/Bcl-2 ratio and active-caspase 3 level, but also down-regulated Sirt1 expression and its activity, as well as PGC-1α expression. Furthermore, compound II also significantly suppressed the promotion of HepG2 cell proliferation, as evidenced by MTT assay and lactate dehydrogenase (LDH) release assay. Of note, the cytotoxicity of Compound II on HepG2 cells mainly regulating Sirt1/caspase 3 signaling pathway, consisting with the results . Intriguingly, z-DEVD-FMK, a caspase 3 inhibitor, almost abolished the inhibitory effects of compound II. Of note, knockdown of caspase 3 by siRNA significantly reversed the inhibitory effect of compound II on HepG2. Interestingly, compound II directly bonded to Sirt1, indicating that Sirt1 might be a promising therapeutic target of compound II. In summary, our findings reveal that compound II, a new synthetical phenyl chlormethine-quinazoline derivative, contributes to the apoptosis of HepG2 cells both and through mediating Sirt1/caspase 3 singling pathway. These findings demonstrate that compound II may be a new potent agent against hepatocellular carcinoma.
喹唑啉衍生物具有多种药理活性，可作用于多种生物受体。基于喹唑啉核心骨架，我们设计并合成了三种带有席夫碱（C = N）连接基团的新型喹唑啉 - 苯基氯甲胺缀合物（I - III），并研究了它们对HepG2异种移植瘤和人肝癌细胞系HepG2的抗肿瘤作用。在这些化合物中，化合物II对HepG2细胞显示出更好的抑制作用。在本研究中，通过TUNEL染色、蛋白质印迹、分子对接和小干扰RNA（siRNA）来研究化合物II对肝癌的抑制机制。化合物II抑制裸鼠体内HepG2异种移植瘤的生长。此外，化合物II不仅上调了Bax/Bcl - 2比值和活化的半胱天冬酶3水平，还下调了沉默信息调节因子1（Sirt1）的表达及其活性以及过氧化物酶体增殖物激活受体γ共激活因子1α（PGC - 1α）的表达。此外，如MTT法和乳酸脱氢酶（LDH）释放试验所示，化合物II还显著抑制了HepG2细胞的增殖促进作用。值得注意的是，化合物II对HepG2细胞的细胞毒性主要通过调节Sirt1/半胱天冬酶3信号通路，这与结果一致。有趣的是，半胱天冬酶3抑制剂z - DEVD - FMK几乎消除了化合物II的抑制作用。值得注意的是，通过siRNA敲低半胱天冬酶3显著逆转了化合物II对HepG2的抑制作用。有趣的是，化合物II直接与Sirt1结合，表明Sirt1可能是化合物II的一个有前景的治疗靶点。总之，我们的研究结果表明，新型合成的苯基氯甲胺 -喹唑啉衍生物化合物II通过介导Sirt1/半胱天冬酶3信号通路促进HepG2细胞凋亡。这些研究结果表明化合物II可能是一种新型强效抗肝细胞癌药物。
