title: Interaction of tetraspan(in) TM4SF5 with CD44 promotes self-renewal and circulating capacities of hepatocarcinoma cells.
abstract: Tumor metastasis involves circulating and tumor-initiating capacities of metastatic cancer cells. Epithelial-mesenchymal transition (EMT) is related to self-renewal capacity and circulating tumor cell (CTC) characteristics for tumor metastasis. Although tumor metastasis is a life-threatening, complicated process that occurs through circulation of tumor cells, mechanistic aspects of self-renewal and circulating capacities have been largely unknown. Hepatic transmembrane 4 L six family member 5 (TM4SF5) promotes EMT for malignant growth and migration, so it was rationalized that TM4SF5, as a hepatocellular carcinoma (HCC) biomarker, might be important for metastatic potential. Here, self-renewal capacity by TM4SF5 was mechanistically explored using hepatocarcinoma cells with or without TM4SF5 expression, and we explored whether they became CTCs using mouse liver-orthotopic model systems. We found that TM4SF5-dependent sphere growth correlated with CD24(-) , aldehyde dehydrogenase (ALDH) activity, as well as a physical association between CD44 and TM4SF5. Interaction between TM4SF5 and CD44 was through their extracellular domains with N-glycosylation modifications. TM4SF5/CD44 interaction activated proto-oncogene tyrosine-protein kinase Src (c-Src)/signal transducer and activator of transcription 3 (STAT3)/Twist-related protein 1 (Twist1)/B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi1) signaling for spheroid formation, whereas disturbing the interaction, expression, or activity of any component in this signaling pathway inhibited spheroid formation. In serial xenografts using 200∼5,000 cells per injection, TM4SF5-positive tumors exhibited subpopulations with locally increased CD44 expressions, supporting for tumor cell differentiation. TM4SF5-positive, but not TM4SF5- or CD44-knocked-down, cells were identified circulating in blood 4-6 weeks after orthotopic liver injection using in vivo laser scanning endomicroscopy. Anti-TM4SF5 reagent blocked their metastasis to distal intestinal organs.
  
  
  CONCLUSIONS
  TM4SF5 promotes self-renewal and CTC properties supported by TM4SF5(+) /CD44(+(TM4SF5-bound)) /ALDH(+) /CD24(-) markers during HCC metastasis.
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Doohyung
authors: D
authors: Lee
authors: Department of Nuclear Medicine, Cancer Research Institute, College of Medicine, Seoul National University, Seoul, Korea.
authors: Juri
authors: J
authors: Na
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Jihye
authors: J
authors: Ryu
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Hye-Jin
authors: HJ
authors: Kim
authors: Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, Korea.
authors: Seo Hee
authors: SH
authors: Nam
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.|Department of Biomedical Sciences, College of Medicine, Seoul National University, Seoul, Korea.
authors: Minkyung
authors: M
authors: Kang
authors: Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, Korea.
authors: Jae Woo
authors: JW
authors: Jung
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Mi-Sook
authors: MS
authors: Lee
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Haeng Eun
authors: HE
authors: Song
authors: Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, Korea.
authors: Jungeun
authors: J
authors: Choi
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Gyu-Ho
authors: GH
authors: Lee
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
authors: Tai Young
authors: TY
authors: Kim
authors: Department of Nuclear Medicine, Cancer Research Institute, College of Medicine, Seoul National University, Seoul, Korea.|Cancer Imaging Center, Seoul National University Hospital, Seoul, Korea.
authors: June-Key
authors: JK
authors: Chung
authors: Division of Applied Life Science, Gyeongsang National University, Jinju, Korea.
authors: Ki Hun
authors: KH
authors: Park
authors: School of Life Sciences and Biotechnology, Korea University, Seoul, Korea.
authors: Sung-Hak
authors: SH
authors: Kim
authors: School of Life Sciences and Biotechnology, Korea University, Seoul, Korea.
authors: Hyunggee
authors: H
authors: Kim
authors: Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon, Korea.
authors: Howon
authors: H
authors: Seo
authors: Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon, Korea.
authors: Pilhan
authors: P
authors: Kim
authors: Department of Nuclear Medicine, Cancer Research Institute, College of Medicine, Seoul National University, Seoul, Korea.|Cancer Imaging Center, Seoul National University Hospital, Seoul, Korea.
authors: Hyewon
authors: H
authors: Youn
authors: Department of Pharmacy, College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.|Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, Korea.
authors: Jung Weon
authors: JW
authors: Lee
mesh_terms: D000818:Animals; D000081247:CSK Tyrosine-Protein Kinase; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D063150:Polycomb Repressive Complex 1 / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D018874:Spheroids, Cellular; D051879:Twist-Related Protein 1 / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: C069077:BMI1 protein, human; C489421:Bmi1 protein, mouse; C497483:CD44 protein, human; D018960:Hyaluronan Receptors; C000715470:MAS1 protein, human; D008565:Membrane Proteins; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D050796:STAT3 Transcription Factor; C111029:TM4SF5 protein, human; D051879:Twist-Related Protein 1; D063150:Polycomb Repressive Complex 1; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human