Purpose

In this study we aimed to assess the anti-tumor effect of co-inhibition of Aurora kinase A (AURKA) and heat shock transcription factor 1 (HSF1) on hepatocellular carcinoma (HCC), as well as to explore the mechanism involved.

Methods

Expression of AURKA and HSF1 in primary HCC tissues and cell lines was detected by immunohistochemistry (IHC), qRT-PCR and Western blotting. AURKA was knocked down in HepG2 and BEL-7402 HCC cells using lentivirus-mediated RNA interference. Next, CCK-8, clone formation, transwell and flow cytometry assays were used to assess their viability, migration, invasion and apoptosis, respectively. The expression of proteins related to cell cycle progression, apoptosis and endoplasmic reticulum stress (ERS) was analyzed using Western blotting. In addition, in vivo tumor growth of HCC cells was assessed using a nude mouse xenograft model, and the resulting tumors were evaluated using HE staining and IHC.

Results

Both AURKA and HSF1 were highly expressed in HCC tissues and cells, while being negatively related to HCC prognosis. Knockdown of AURKA significantly inhibited the colony forming and migrating capacities of HCC cells. In addition, we found that treatment with an AURKA inhibitor (Danusertib) led to marked reductions in the proliferation and migration capacities of the HCC cells, and promoted their apoptosis. Notably, combined inhibition of AURKA and HSF1 induced HCC cell apoptosis, while increasing the expression of ERS-associated proteins, including p-eIF2α, ATF4 and CHOP. Finally, we found that co-inhibition of AURKA and HSF1 elicited an excellent in vivo antitumor effect in a HCC mouse model with a relatively low cytotoxicity.

Conclusions

Combined inhibition of AURKA and HSF1 shows an excellent anti-tumor effect on HCC cells in vitro and in vivo, which may be mediated by ERS. These findings suggest that both AURKA and HSF1 may serve as targets for HCC treatment.

中文翻译：

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AURKA和HSF1联合抑制通过激活内质网应激抑制肝癌增殖并促进细胞凋亡

目的

在本研究中，我们旨在评估 Aurora 激酶 A (AURKA) 和热休克转录因子 1 (HSF1) 共同抑制肝细胞癌 (HCC) 的抗肿瘤作用，并探讨相关机制。

方法

通过免疫组织化学 (IHC)、qRT-PCR 和蛋白质印迹检测 AURKA 和 HSF1 在原发性 HCC 组织和细胞系中的表达。使用慢病毒介导的 RNA 干扰在 HepG2 和 BEL-7402 HCC 细胞中敲低 AURKA。接下来，CCK-8、克隆形成、transwell 和流式细胞仪检测分别用于评估它们的活力、迁移、侵袭和凋亡。使用蛋白质印迹分析与细胞周期进程、细胞凋亡和内质网应激 (ERS) 相关的蛋白质的表达。此外，使用裸鼠异种移植模型评估 HCC 细胞的体内肿瘤生长，并使用 HE 染色和 IHC 评估所得肿瘤。

结果

AURKA 和 HSF1 在 HCC 组织和细胞中均高表达，而与 HCC 预后呈负相关。敲除 AURKA 显着抑制了 HCC 细胞的集落形成和迁移能力。此外，我们发现用 AURKA 抑制剂 (Danusertib) 治疗导致 HCC 细胞的增殖和迁移能力显着降低，并促进了它们的凋亡。值得注意的是，AURKA 和 HSF1 的联合抑制可诱导 HCC 细胞凋亡，同时增加 ERS ​​相关蛋白的表达，包括 p-eIF2α、ATF4 和 CHOP。最后，我们发现 AURKA 和 HSF1 的共同抑制在具有相对较低细胞毒性的 HCC 小鼠模型中引发了出色的体内抗肿瘤作用。

结论

AURKA 和 HSF1 的联合抑制在体外和体内对 HCC 细胞显示出优异的抗肿瘤作用，这可能是由 ERS ​​介导的。这些发现表明 AURKA 和 HSF1 都可以作为 HCC 治疗的靶点。