A new 1,2,3-triazole-indirubin hybrid suppresses tumor growth and pulmonary metastasis by mitigating the HGF/c-MET axis in hepatocellular carcinoma.

Shalini V Gowda, Na Young Kim, Kachigere B Harsha, Darshini Gowda, Rajaghatta N Suresh, Amudha Deivasigamani, Chakrabhavi Dhananjaya Mohan, Kam Man Hui, Gautam Sethi, Kwang Seok Ahn, Kanchugarakoppal S Rangappa
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引用次数: 0

Abstract

Introduction: Hepatocellular carcinoma (HCC) is a fatal cancer that is often diagnosed at the advanced stages which limits the available therapeutic options. The interaction of HGF with c-MET (a receptor tyrosine kinase) results in the activation of c-MET which subsequently triggers the PI3K/Akt/mTOR axis. Overexpression of c-MET in HCC tissues has been demonstrated to contribute to tumor progression and metastasis.

Objectives: We aimed to synthesize triazole-indirubin conjugates, examine their growth suppressor efficacy in cell-based assays, and investigate the antitumor as well as antimetastatic activity of lead cytotoxic agent in the orthotopic mice model.

Methods: A series of triazole-indirubin hybrids were synthesized and cytotoxicity, apoptogenic, and antimigratory effect of the lead compound (CRI9) was evaluated using MTT assay, cell cycle analysis, annexin-V/PI assay, TUNEL assay, and wound healing assay. The effect of CRI9 on the operation of the HGF/c-MET/PI3K/Akt/mTOR axis was examined using western blotting and transfection experiments. Acute toxicity, antitumor, and antimetastatic activity of CRI9 were examined in NCr nude mice. The expression of c-MET/PI3K/Akt/mTOR, CD31, and Ki-67 was examined using immunohistochemistry and western blotting.

Results: Among the new compounds, CRI9 consistently displayed potent cytotoxicity against HGF-induced HCC cells. CRI9 induced apoptosis as evidenced by increased sub G1 cells, annexin-V+/PI+ cells, TUNEL+ cells, and cleavage of procaspase-3 and PARP. CRI9 inhibited HGF-induced phosphorylation of c-METY1234/1235 and subsequently suppressed the PI3K/Akt/mTOR axis. Also, depletion of c-MET or inhibition of c-MET by CRI9 resulted in suppression of the PI3K/Akt/mTOR axis. CRI9 showed no toxic effects in NCr nude mice and displayed a potent antitumor and antimetastatic effect in the orthotopic HCC mice model. CRI9 also reduced the levels of phospho-c-MET, CD31, and Ki-67 and suppressed the activation of the PI3K/Akt/mTOR axis in tumor tissues.

Conclusion: CRI9 has been identified as a new inhibitor of the c-MET/PI3K/Akt/mTOR axis in HCC preclinical models.

一种新型 1,2,3-三唑-靛红混合物通过缓解肝细胞癌中的 HGF/c-MET 轴抑制肿瘤生长和肺转移。
简介肝细胞癌(HCC)是一种致命的癌症,通常在晚期才被诊断出来,这限制了现有的治疗方案。HGF与c-MET(一种受体酪氨酸激酶)的相互作用会导致c-MET的活化,进而触发PI3K/Akt/mTOR轴。c-MET 在 HCC 组织中的过表达已被证实有助于肿瘤的进展和转移:目的:我们旨在合成三唑-靛红共轭物,在基于细胞的实验中检验其抑制生长的功效,并在正位小鼠模型中研究主要细胞毒剂的抗肿瘤和抗转移活性:方法:通过多步反应合成了新的三唑-靛红杂交化合物。方法:通过多步反应合成了新的三唑-靛红杂交化合物,并使用 MTT 试验、细胞周期分析、annexin-V/PI 试验、TUNEL 试验和伤口愈合试验评估了新化合物(CRI9)的细胞毒性、致凋亡和抗移植物作用。CRI9对HGF/c-MET/PI3K/Akt/mTOR轴运行的影响通过Western印迹和转染实验进行了检验。在NCr裸鼠体内检测了CRI9的急性毒性、抗肿瘤和抗转移活性。免疫组化和免疫印迹法检测了c-MET/PI3K/Akt/mTOR、CD31和Ki-67的表达:结果:在这些新化合物中,CRI9对HGF诱导的HCC细胞始终显示出强大的细胞毒性。CRI9可诱导细胞凋亡,表现为亚G1细胞、annexin-V+/PI+细胞、TUNEL+细胞的增加,以及procaspase-3和PARP的裂解。CRI9 可抑制 HGF 诱导的 c-METY1234/1235 磷酸化,进而抑制 PI3K/Akt/mTOR 轴。此外,CRI9消耗c-MET或抑制c-MET也会抑制PI3K/Akt/mTOR轴。CRI9对NCr裸鼠无毒性作用,在正位HCC小鼠模型中显示出强大的抗肿瘤和抗转移作用。CRI9还能降低肿瘤组织中磷酸化c-MET、CD31和Ki-67的水平,抑制PI3K/Akt/mTOR轴的激活:结论:在 HCC 临床前模型中,CRI9 被确定为一种新的 c-MET/PI3K/Akt/mTOR 轴抑制剂。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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