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Ras association domain family member 10 suppresses gastric cancer growth by cooperating with GSTP1 to regulate JNK/c-Jun/AP-1 pathway

Oncogene volume 35, pages 2453–2464 (2016)Cite this article

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Abstract

The Ras association domain family (RASSF) encodes several members with tumor-suppressive potentials. We aimed to investigate the biological function and clinical implication of RASSF10 in gastric cancer (GC). We found that RASSF10 was silenced in six of seven GC cell lines and in primary GC tissues, but was highly expressed in normal gastric tissues. The silence of RASSAF10 was mediated by promoter methylation as evaluated by bisulfite genomic sequencing. RASSF10 expression could be restored by demethylation treatment. A negative correlation between methylation and mRNA expression of RASSF10 was observed in 223 gastric samples of The Cancer Genome Atlas study (P<0.0001). Re-expression of RASSF10 in GC cell lines (AGS and MKN45) significantly suppressed cell viability, colony formation, migration and invasion, reduced cells in S phase, accumulated cells in G2 phase and induced cell apoptosis in vitro, and inhibited tumorigenicity in nude mice. These were confirmed by decreased expression of proliferation markers (proliferating cell nuclear antigen, p-CDC2 and p-CDC25) and increased apoptotic cascades (cleaved caspases-9, -8, -3 and cleaved poly (ADP-ribose) polymerase). Conversely, RASSF10 knockdown in normal gastric cell line yielded an opposing effect. Co-immunoprecipitation combined with mass spectrometry analyses were performed to reveal the downstream effectors of RASSF10. The result revealed that glutathione S-transferase Pi 1 (GSTP1) was a direct cooperator of RASSF10. The tumor-suppressive effect of RASSF10 was partially mediated by cooperating with GSTP1 to deregulate Jun N-terminal kinase (JNK)/c-Jun/AP-1 pathway. Importantly, RASSF10 methylation was detected in 56.6% (98/173) of primary GCs and is an independent risk factor for poor survival of GC patients (P=0.001). In conclusions, RASSF10 functions as a tumor suppressor by cooperating with GSTP1 to deregulate JNK/c-Jun/AP-1 pathway in GC. Promoter methylation of RASSF10 is associated with poor survival of GC patients.

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Abbreviations

BGS:

bisulfite genomic sequencing

GC:

gastric cancer

GSTP1:

glutathione S-transferase Pi 1

JNK:

Jun N-terminal kinase

MS-RDA:

methylation-sensitive representational difference analysis

RASSF10:

Ras association domain family member 10

RT–PCR:

reverse transcription–PCR

shRNA:

small hairpin RNA

siRNA:

short interference RNA

TUNEL:

terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling.

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Acknowledgements

This project was supported by research funds of Natural Science Foundation of China (NSFC) (81201963 and 81372600), a China 863 Program (2012AA02A504), Shenzhen Municipal Science and Technology R&D fund (JCYJ20130401151108652), Pearl River S&T Nova Program of Guangzhou (201506010050) and Shenzhen Virtual University Park Support Scheme to CUHK Shenzhen Research Institute. The results shown here are in part based on data generated by the TCGA Research Network: http://cancergenome.nih.gov/.

Author contributions

XL, WL, YZ, NZ, LX, XZ and JZ performed the experiments. XL and QL analyzed data and drafted the paper. JJS commented on the study. JY designed, supervised study and revised the paper.

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Authors and Affiliations

  1. Institute of Digestive Disease and Department of Medicine and Therapeutics, State Key Laboratory of Digestive Disease, Li Ka Shing Institute of Health Sciences, CUHK Shenzhen Research Institute, The Chinese University of Hong Kong, Hong Kong, China

    X Li, Q Liang, L Xu, X Zhang, J Zhang, J J Y Sung & J Yu

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

    X Li

  3. Department of Gastroenterology, Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou, China

    W Liu

  4. Department of Gastroenterology, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China

    N Zhang & L Xu

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Correspondence to J Yu.

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Li, X., Liang, Q., Liu, W. et al. Ras association domain family member 10 suppresses gastric cancer growth by cooperating with GSTP1 to regulate JNK/c-Jun/AP-1 pathway. Oncogene 35, 2453–2464 (2016). https://doi.org/10.1038/onc.2015.300

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