Understanding functional roles of FAM86A in NF-κB mediated gastric tumorigenic responses 2 1 1 1 1 1 1 1 Yo Han Hong , Jae Gwang Park , Han Gyung Kim , Nur Aziz , Long You , Chaoran Song , Wooram Choi , Jieun Oh and Jae Youl Cho 1 1 Integrative biotechnology, Sungkyunkwan University, Suwon 16419, Korea 2 Colorectal Cancer Branch, National Cancer Center, Goyang 10408, Korea BACKGROUND AIM Gastric cancer is the one of the most lethal and third In this study, we investigated functional roles of cause of cancer related mortality. Gastric cancer is FAM86A on the gastric tumorigenic responses.
involved in various oncogenic signaling pathways. FGFR1-PI3K-AKT-NF-κB signaling pathway affects the METHODS expression of various genes that regulate tumorigenic responses. These responses are known to be regulated Clinical impact of FAM86A was assessed in total 426 patients with gastric cancer. Immunoblot and realtime PCR by post-translational modifications. Methylation as one of were used to detect FAM86A expression in cell lines and the post-translational modifications change protein patient samples. QunatSeq 3’ mRNA-Seq was used to structure, activity, and localization. FAM86A as a non-
screen the target genes of FAM86A. Biological functions of histone protein lysine methyltransferase has novelty and FAM86A were examined in vitro and in vivo. conserved sequence in the various species. However, Immunoprecipitation and mass spectrometry were biological functions of FAM86A remain unclear. conducted to identify the specific binding partners. RESULTS Here, we report the identification of the Figure 1. Clinicopathological Figure 2. Biological landscape of FAM86A as a candidate biomarker in gastric significance of FAM86A in gastric FAM86A cancer. FAM86A was overexpressed in human
cancer gastric cancer specimens and cell lines. Using gastric cancer patient samples from Asia Cancer Research Group (ACRG) database and Ajou University Hospital, FAM86A overexpression positively correlated with clinicopathological characteristics of gastric cancer such as lymph vascular invasion and Lauren intestinal type, and was independently associated with poor survival. RNA sequencing analysis revealed that knockdown of FAM86A Figure 4. Molecular mechanism of in gastric cancer cells is accompanied by a Figure 3. Oncogenic function of FAM86A decrease in the expression of angiogenesis,
FAM86A in vitro and in vivo metastasis and chemotaxis related genes, including VEGF, IL-6 and MMP7. Results from loss-of-function and gain-of-function experiments suggested that knockdown of FAM86A attenuated, while overexpression of FAM86A enhanced, the invasion, migration and colony forming ability of gastric cancer cells in vitro and in vivo. Mechanistically, we found that FGFR1-PI3K-AKT-NF-κB signal pathway and its downstream effectors, such as MMP7, VEGF and IL-6, were required for FAM86A mediated tumor metastasis of gastric cancer. FAM86A interacted with FGFR1 and subsequently activated
the FGFR1-PI3K-AKT- Figure 5. Clinical relevance NF-κB signaling pathway by methylation between FAM86A and FGFR1 dependent manner. We also found that protein expression of FAM86A was correlated with activation of FGFR1 in human gastric cancer specimens. CONCLUSION REFERENCES ACKNOWLEDGEMENTS [1] HAMAMOTO, Ryuji; SALOURA, Vassiliki; NAKAMURA, This work was supported by National Research Foundation Yusuke. Critical roles of non-histone protein lysine methylation of Korea (NRF) funded by the Korean Government (NRF- This study suggests that in human tumorigenesis. Nature Reviews Cancer, 2015,
15.2: 2016H1A2A1908921 Global Ph.D. Fellowship Program) and upregulation of FAM86A results in 110-124. Ministry of Education (NRF-2018R1D1A1B07047970 Basic Science Research Program) gastric cancer, leading to [2] DAVYDOVA, Erna, et al. Identification and characterization lysine-specific conserved a of evolutionarily novel metastasis via activation of the methyltransferase targeting eukaryotic translation elongation Contact information factor 2 (eEF2). Journal of Biological Chemistry, 2014, 289.44: FGFR1-PI3K-AKT-NF-κB signaling 30499-30510. pathway. [3] CRISTESCU, Razvan, et al. Molecular
analysis of gastric Yo Han Hong: ghddygks12@skku.edu cancer identifies subtypes associated with distinct clinical Jae Youl Cho: jaecho@skku.edu outcomes. Nature medicine, 2015, 21.5: 449. [D. Cancer biology-1] Understanding functional roles of FAM86A in NF-kB mediated gastric tumorigenic responses Yo Han Hong¹, Jieun Oh¹, Wooram Choi¹, Han Gyung Kim¹, Deok Jeong¹, Chaoran Song¹, Long You¹, Jae Youl Cho¹˙* ¹Integrative biotechnology, Sungkyunkwan University, Suwon 16419, Korea FAM86A as a non-histone protein lysine methyltransferase has novelty and conserved sequence in the various species. In
this study, we investigated functional roles of FAM86A on the gastric tumorigenic responses. According to microarray analysis against Asian Cancer Research Group, FAM86A was significantly overexpressed in tumor compared to normal. Also, FAM86A overexpressed gastric cancer patient’s prognosis was known to be worse than control group. FAM86A was occurred genetic alteration in the cancer patients. FAM86A was overexpressed in most gastric tumor tissue and cancer cell lines. Moreover, FAM86A involved in various tumorigenic responses including cell proliferation, cell cycle, invasion, migration and
colony formation. And then, we figured out the mechanism by which FAM86A regulates gastric tumorigenic responses. As results, FAM86A activated FGFR1, a novel binder partner of FAM86A, mediated NF-kB signaling pathway by the methylation dependent manner. FAM86A regulated tumor growth and metastasis in vivo mice model. These results suggest that FAM86A could regulate gastric tumorigenic responses through FGFR1 mediated NF-kB signaling pathway by the methylation dependent manner. Metastatic function of METTL18 in breast cancer via actin methylation and Src 1 1 Jieun Oh , Han Gyung Kim , Yo Han
Hong , Wooram Choi , Deok Jeong , Chaoran Song , Long You , and Jae Youl Cho 1 1 1 1 1 1 1 Department of Integrative Biotechnology, Sungkyunkwan University, Suwon, 16419, Republic of Korea BACKGROUND AIM Breast cancer is the most common form of malignant tumor and We examined methylation of histidine (His)-73 residue of actin by second leading cause of cancer-related deaths in women. Triple METTL18 as a crucial component of metastatic responses of breast tumor cells through modulation of the actin cytoskeleton and negative breast cancer (TNBC), accounts for 10 - 15 % of breast Src
phosphorylation. cancer and neither of approved drugs nor established chemotherapy targeting TNBC at present. METTL18 (histidine METHODS methyltransferase 1 homolog) is considered a putative human homolog of YY110W (yeast histidine methyltransferase). B-actin, Immunoblot and realtime PCR were used to detect METTL18 S100A9, myosin, myosin kinase, and ribosomal protein RPL3 expression in cell lines and patient samples. Biological functions of were known to be methylated at histidine residues and may be METTL18 were examined in vitro and in vivo. Via tumor xenografts, quite common in
intracellular proteins in mammalian cells. metastasis assay and PET/CT, effect of METTL18 in metastatic However, biological function and molecular characterization of response were analyzed. Immunoprecipitation and mass METTL18 have yet to be elucidated. spectrometry were conducted to determine the methylation of actin histidine73 residue. RESULTS Here, in this study, we reported another type of actin histidine methyltransferase, METTL18, regulates the metastatic potential of breast cancer in human. Among methyltransferases, METTL18 was highly amplified in human breast cancer. In particular,
overexpressed METTL18 showed poor prognosis. Knockdown of METTL18 significantly reduced metastatic responses of breast tumor cells both in vitro and in vivo. Mechanistically, it was observed that METTL18 increased actin polymerization, upregulated complex formation with HSP90AA1 and Src, enhanced the activity of an intermediate form of Src with tyrosine phosphorylation at both Y416 and Y527, and induced cellular metastatic responses, including morphological change, migration, and invasion of MDA-MB231 cells in vitro and in vivo. Methylated actin at His73 served as a critical site for
interaction with HSP90AA1 and Src to activate p85/PI3K and STAT3. CONCLUSION REFERENCES ACKNOWLEDGEMENTS This research was supported by the Basic Science [1] Camara-Artigas A, Martinez-Rodriguez S, Ortiz-Salmeron E, Martin-Garcia JM (2014) 3D domain swapping in a chimeric Research Program through the National Research c-Src SH3 domain takes place through two hinge loops. Foundation of Korea (NRF) funded by the Ministry Journal of structural biology 186: 195-203 of Education (2017R1A6A1A03015642,) Republic of METTL18 regulates tumorigenic [2] Carpenter CL (2000) Actin cytoskeleton and cell
Korea signaling. Crit Care Med 28: N94-9 response of breast cancer through [3] Cloutier P, Lavallee-Adam M, Faubert D, Blanchette M, Contact information Coulombe B (2013) A newly uncovered group of distantly methylation and polymerization of related lysine methyltransferases preferentially interact actin with HSP90AA1 with molecular chaperones to regulate their activity. PLoS Jae Youl Cho : jaecho@skku.edu Genet 9: e1003210 [4] Gonzalez L, Agullo-Ortuno MT, Garcia-Martinez JM, Calcabrini A, Gamallo C, Palacios J, Aranda A, Martin-Perez J Jieun Oh : martia96@gmail.com (2006) Role of c-Src in
human MCF7 breast cancer cell tumorigenesis. J Biol Chem 281: 20851-64 Han Gyung Kim: hanks523@naver.com [D. Cancer biology-2] Metastatic function of METTL18 in breast cancer via actin methylation and Src Jieun Oh¹˙#, Han Gyung Kim¹˙#, Yo Han Hong¹, Wooram Choi¹, Deok Jeong¹, Chaoran Song¹, Long You¹, Jae Youl Cho¹˙* ¹Department of Integrative Biotechnology, Sungkyunkwan University, Suwon 16419, Korea In this study, we report another type of actin histidine methyltransferase. METTL18 regulates the metastatic potential of breast cancer in human. Among methyltransferases, METTL18 was highly
amplified in human breast cancer. In particular, poor prognosis was associated with high expression of METTL18 in HER2-negative breast cancer patients. This gene product was also found to be a critical component of metastatic responses. Loss of METTL18 expression significantly reduced metastatic responses of breast tumor cells both in vitro and in vivo. Mechanistically, it was observed that METTL18 increased actin polymerization, upregulated complex formation with HSP90AA1 and Src, enhanced the activity of an intermediate form of Src with tyrosine phosphorylation at both Y416 and Y527, and
induced cellular metastatic responses, including morphological change, migration, and invasion of MDA-MB-231 cells in vitro and in vivo. Methylated actin at His73 served as a critical site for interaction with HSP90AA1 and Src to activate p85/PI3K and STAT3. Our findings suggest that METTL18 plays critical roles in metastatic responses of HER2-negative breast cancer cells via actin polymerization and the generation of an intermediate form of Src. Regulatory mechanism of MKK7 by METTL21B in gastric cancer Long You , Yo Han Hong , Jae Gwang Park , Nur Aziz , Chaoran Song , Wooram Choi , Jieun Oh
, Han Gyung Kim , and Jae Youl Cho 1 1 1 2 1 1 1 1 1 1 Integrative biotechnology, Sungkyunkwan University, Suwon 16419, Korea 2 Colorectal Cancer Branch, National Cancer Center, Goyang 10408, Korea BACKGROUND AIM Gastric cancer is difficult to diagnose early because there To figure out clinical relevance and biological are no symptoms at the beginning of development, and significance of METTL21B in human gastric cancer. most of the advanced cancer is diagnosed as advanced cancer, and treatment methods are very limited and poor. METHODS Mitogen-activated protein kinase pathway is a major cell-
mediated cascade that regulates processes that regulate Clinical im
