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Elevated expression of RIT1 hyperactivates RAS/MAPK signal and sensitizes hepatocellular carcinoma to combined treatment with sorafenib and AKT inhibitor

Oncogene volume 41, pages 732–744 (2022)Cite this article

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Abstract

Hyperactivation of RAS/MAPK signaling is commonly observed in hepatocellular carcinoma (HCC). Gain-of-function mutations of canonical RAS genes, however, are rarely detected and it remains unclear how the activity of this pathway is turned on during hepatocarcinogenesis. We performed a comprehensive analysis of RAS superfamily genetic alterations across ten subfamilies, 152 members in 377 HCC patients from the Cancer Genome Atlas database. RIT1 (Ras-like without CAAX 1) was the most frequently altered RAS member amplified in 13% of the HCC cohort. Both genomic amplification and CREB-mediated transcriptional activation contributed to the elevated RIT1 expression, and its overexpression correlated with RAS/MAPK activation and poor prognosis. Then, we found that RIT1-induced angiogenesis via the MEK/ERK/EIF4E/HIF1-α/VEGFA axis. MAP3K11 and MAP3K12, in addition to CRAF, could mediate this process by binding to RIT1. Moreover, RIT1 increased the phosphorylation of p38 MAPK and AKT to promote cell survival under reactive oxygen species stress. Based on this mechanistic understanding, we treated RIT1-overexpressing HCC with combined regimen sorafenib plus AKT inhibitor, and achieved enhanced antitumor effects in vivo. Our study reveals RAS “orphan” member RIT1 as the most common genetic alteration of RAS family in HCC and combination of sorafenib with AKT inhibitor might be a promising treatment strategy for RIT1-overexpressing HCC.

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Fig. 1: Copy number amplification of RIT1 is the most frequent genetic alteration of RAS family in HCC and associates with RAS/MAPK activation.
Fig. 2: RIT1 overexpression can be transcriptionally activated by CREB protein and associates with poor prognosis.
Fig. 3: RIT1 confers robust tumorigenic and metastatic properties to HCC cells.
Fig. 4: RIT1 promotes a tumor vasculature phenotype.
Fig. 5: RIT1 induces angiogenesis through upregulation of VEGFA.
Fig. 6: RIT1 activates MEK/ERK via multiple MAP3Ks.
Fig. 7: RIT1 enhances HCC cell survival and metastasis under ROS stress.
Fig. 8: RIT1 is a useful biomarker to classify patients for sorafenib plus AKT inhibitor combined treatment.

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Acknowledgements

The authors would like to thank the Core Facility of University of Hong Kong for their support for flow cytometry analysis, confocal microscopy, and animal imaging.

Funding

This work was supported by the National Natural Science Foundation of China (82073127), Shenzhen Fundamental Research Programs (JCYJ20190809145215160), Shenzhen Science and Technology Innovation Commission (KYTDPT20181011104005 and KQTD2018041118502879), and Guangdong Innovative Research Team Fund (No. 2016ZT06S172). Hong Kong Research Grant Council grants including GRF (17143716), Collaborative Research Funds (C7065-18GF and C7026-18GF) and Theme-based Research Scheme (T12-704/16-R).

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Author notes

  1. These author contributed equally: Xin-Yuan Guan, Yan Li.

Authors and Affiliations

  1. Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China

    Liangzhan Sun, Zhengdong Zhou, Pengchao Hu, Shasha Wu, Yanchen Wang, Yuyang Du, Jingyi Zheng, Hui Yang, Dandan Yu & Yan Li

  2. Department of Clinical Oncology, The University of Hong Kong, Hong Kong, China

    Liangzhan Sun, Feifei Zhang, Yuzhu Cui, Shayi Wu, Miao Chen, Qian Yan, Dandan Yu, Chaoran Shi, Yu Zhang & Xin-Yuan Guan

  3. State Key Laboratory for Liver Research, The University of Hong Kong, Hong Kong, China

    Liangzhan Sun, Qian Yan, Dandan Yu & Xin-Yuan Guan

  4. Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, China

    Shaoyan Xi

  5. State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, China

    Shaoyan Xi, Ying Wang, Dan Xie & Xin-Yuan Guan

  6. Department of Radiation Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

    Ying Wang

  7. University of Hong Kong-Shenzhen Hospital, Shenzhen, China

    Xin-Yuan Guan

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Contributions

XY-G, YL and LS devised and coordinated the project. LS performed all the experiments with help from SX, ZZ, FZ, PH, YC, ShashaW, YingW, ShayiW, YanchenW, YD, JZ, HY, MC, QY, DY, CS, YZ. DX and SX provided human samples, performed tumor grades and histological examination. LS and YL wrote the manuscript. XY-G and YL supervised the project.

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Correspondence to Xin-Yuan Guan or Yan Li.

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Sun, L., Xi, S., Zhou, Z. et al. Elevated expression of RIT1 hyperactivates RAS/MAPK signal and sensitizes hepatocellular carcinoma to combined treatment with sorafenib and AKT inhibitor. Oncogene 41, 732–744 (2022). https://doi.org/10.1038/s41388-021-02130-8

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