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

当前位置： X-MOL 学术 › J. Adv. Res. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

A new 1,2,3-triazole-indirubin hybrid suppresses tumor growth and pulmonary metastasis by mitigating the HGF/c-MET axis in hepatocellular carcinoma
Journal of Advanced Research ( IF 11.4 ) Pub Date : 2024-08-30 , DOI: 10.1016/j.jare.2024.08.033
Shalini V Gowda ₁ , Na Young Kim ₂ , Kachigere B Harsha ₁ , Darshini Gowda ₁ , Rajaghatta N Suresh ₁ , Amudha Deivasigamani ₃ , Chakrabhavi Dhananjaya Mohan ₄ , Kam Man Hui ₃ , Gautam Sethi ₅ , Kwang Seok Ahn ₂ , Kanchugarakoppal S Rangappa ₁

Affiliation

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-MET^Y1234/1235 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.

中文翻译：

一种新的 1,2,3-三唑-靛玉红杂交体通过减轻肝细胞癌中的 HGF/c-MET 轴来抑制肿瘤生长和肺转移

介绍

肝细胞癌（HCC）是一种致命的癌症，通常在晚期被诊断出来，这限制了可用的治疗选择。HGF 与 c-MET（一种受体酪氨酸激酶）的相互作用导致 c-MET 激活，随后触发 PI3K/Akt/mTOR 轴。HCC 组织中 c-MET 的过表达已被证明会导致肿瘤进展和转移。

目标

我们旨在合成三唑-靛玉红偶联物，在基于细胞的测定中检查其生长抑制因子的功效，并研究铅细胞毒剂在原位小鼠模型中的抗肿瘤和抗转移活性。

方法

合成一系列三唑-靛玉红杂交体，并使用 MTT 测定、细胞周期分析、膜联蛋白-V/PI 测定、TUNEL 测定和伤口愈合测定评价先导化合物（CRI9）的细胞毒性、凋亡和抗迁移作用。使用 western blotting 和转染实验检测 CRI9 对 HGF/c-MET/PI3K/Akt/mTOR 轴操作的影响。在 NCr 裸鼠中检测 CRI9 的急性毒性、抗肿瘤和抗转移活性。使用免疫组化和蛋白质印迹检测 c-MET/PI3K/Akt/mTOR 、 CD31 和 Ki-67 的表达。

结果

在新化合物中，CRI9 始终对 HGF 诱导的 HCC 细胞表现出强大的细胞毒性。CRI9 诱导细胞凋亡，表现为 sub G1 细胞、膜联蛋白-V⁺/PI⁺ 细胞、TUNEL⁺ 细胞增加以及 procaspase-3 和 PARP 裂解。CRI9 抑制 HGF 诱导的 c-MET^Y1234/1235 磷酸化，随后抑制 PI3K/Akt/mTOR 轴。此外，c-MET 的耗竭或 CRI9 对 c-MET 的抑制导致 PI3K/Akt/mTOR 轴的抑制。CRI9 在 NCr 裸鼠中没有毒性作用，在原位 HCC 小鼠模型中显示出有效的抗肿瘤和抗转移作用。CRI9 还降低了磷酸化 c-MET 、 CD31 和 Ki-67 的水平，并抑制了肿瘤组织中 PI3K/Akt/mTOR 轴的激活。