KIAA1199 promotes sorafenib tolerance and the metastasis of hepatocellular carcinoma by activating the EGF/EGFR-dependent epithelial-mesenchymal transition program

doi:10.1016/j.canlet.2019.03.049

Cancer Letters

Volume 454, 10 July 2019, Pages 78-89

https://doi.org/10.1016/j.canlet.2019.03.049 Get rights and content

Highlights

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Sorafenib-resistant HCC xenografts models were established to illustrate sorafenib resistance in HCC.
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KIAA1199 was positively correlated to high risk of recurrence and metastasis in HCC patients.
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KIAA1199 enhanced the metastasis of sorafenib-resistant HCC cells by activation of EGF/EGFR-dependent EMT programs.

Abstract

Patients with advanced hepatocellular carcinoma (HCC) will almost always develop acquired tolerance after sorafenib therapy, and the molecular mechanism of sorafenib tolerance remains poorly characterized. Here, using our established sorafenib-resistant HCC cell and xenograft models, we identified a novel gene, KIAA1199, which was markedly elevated among the differentially expressed genes involved in sorafenib tolerance. Moreover, elevated expression of KIAA1199 was positively correlated with a high risk of recurrence and metastasis and advanced TNM stage in HCC patients. Functionally, loss- and gain-of-function studies showed that KIAA1199 promoted the migration, invasion, and metastasis of sorafenib-resistant HCC cells. Mechanistically, KIAA1199 is required for EGF-induced epithelial-mesenchymal transition (EMT) in sorafenib-resistant HCC cells by aiding in EGFR phosphorylation. In summary, our data uncover KIAA1199 as a novel sorafenib-tolerant promoting gene that plays an indispensable role in maintaining sorafenib-resistant HCC cell metastasis.

Introduction

Hepatocellular carcinoma (HCC) is one of the most prevalent and aggressive neoplasms around the world and the second leading cause of cancer death in developing countries and the sixth leading cause in developed countries [1]. So far, the prognosis of patients with HCC remains dismal, and its overall 5-year survival is less than 15 % [2]. In general, a late diagnosis, high heterogeneous background and HCC cells tolerant to conventional chemotherapeutic agents are considered the principle reasons for the high mortality rates of HCC [3]. However, the molecular and genetic events underlying drug tolerance and tumor metastasis have not been elucidated.

Currently, sorafenib is the first targeted therapy for advanced HCC [4]. Sorafenib mainly blocks tumor cell proliferation by targeting Raf/MEK/ERK signaling and exerts an antiangiogenic effect by VEGFR 2/3 as well as PDGFR-β tyrosine kinases [5]. Although previous data have shown that the clinical outcome of sorafenib for advanced HCC is notable, unfortunately, long-term exposure to sorafenib often leads to tumor cells acquiring drug tolerance [[6], [7], [8]]. To date, the precise molecular mechanism of sorafenib tolerance remains poorly characterized. Thus, the identification of critical targets for the modulation of HCC drug tolerance and migration is valuable for the prevention of tumor cell dissemination and metastasis.

Acquired sorafenib resistance develops in the background of the tumor-host interaction. To identify novel and key targets involved in sorafenib tolerance, we established several sorafenib-resistant HCC cell and xenograft models. The HCC xenograft model more closely resembles the clinical features of HCC patients and retains the characteristics significantly associated with drug response [9].

In our current study, we found that the expression of KIAA1199 was significantly elevated in our previously established sorafenib-resistant HCC in vitro cell model and in an in vivo xenograft model. Furthermore, we found that KIAA1199 promotes HCC cell resistance to sorafenib by enhancing HCC cell migration and invasion. In addition, KIAA1199 promotes HCC cell resistance to sorafenib and cell migration through modulating the EGF-induced epithelial-mesenchymal transition (EMT) process. Our study suggests that KIAA1199 is a novel candidate target for sorafenib tolerance and metastasis in HCC.

Section snippets

Cell culture and reagents

Human HCC cell lines PLC/PRF/5 and HepG2 were purchased from the ATCC (https://www.atcc.org/). Huh7 and HLE cells were obtained from the cell bank at the Chinese Academy of Sciences (Shanghai, China, http://www.cellbank.org.cn/). 293T cells were purchased from the ATCC for lentivirus production. T421, T1115 and T1224 primary HCC cells were previously established in our laboratory. Cells were cultured according to the providers’ instructions for less than 2 months and confirmed by mycoplasma

In vivo establishment of sorafenib-resistant HCC xenograft models

First, we developed several sorafenib-resistant in vivo xenograft models of HCC based on our previously established sorafenib-resistant HCC in vitro cell models [10]. Of note, we systematically assessed sorafenib-resistant HCC cells in vitro before use in in vivo studies. Expectedly, sorafenib-resistant HCC cells possess slow growth characteristics (Supplementary Fig. S1A) and require higher doses of sorafenib for partial inhibition in vitro (Supplementary Fig. S1B). More importantly, in

Discussion

Sorafenib is the first and only targeted therapy for advanced HCC [14]. However, the emergence of acquired sorafenib tolerance has prevented patients from benefiting from this drug [15,16]. At present, research on drug resistance is mainly focused on the natural resistance or induced resistance of tumor cell lines and often leads to a tumor centric drug resistance mechanism [17]. However, tumor-host interactions cannot be reflected in vitro using drug resistance models. Previous reports have

Conflicts of interest

All authors declare no conflicts of interest.

Acknowledgements

This work was supported by funds from National Natural Science Foundation of China (No. 81525020 and 31570753 to J.L., No. 81330048 and 81520108025 to C.Q., No. 81301864 to Y.X.) and the National Key Research and Development Program of China (2016YFC1303405, to C.Q.).

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Original ArticlesKIAA1199 promotes sorafenib tolerance and the metastasis of hepatocellular carcinoma by activating the EGF/EGFR-dependent epithelial-mesenchymal transition program