Development of suspension cell culture model to mimic circulating tumor cells

// Ji Young Park 1, * , Ae Lee Jeong 1, * , Hyun Jeong Joo 1 , Sora Han 1 , So-Hyun Kim 2 , Hye-Youn Kim 2 , Jong-Seok Lim 1 , Myeong-Sok Lee 1 , Hyung-Kyoon Choi 2 and Young Yang 1 1 Department of Biological Sciences, Sookmyung Women’s University, Seoul 04312, Republic of Korea 2 College of Pharmacy, Chung-Ang University, Seoul 06974, Republic of Korea * These authors contributed equally to this work Correspondence to: Hyung-Kyoon Choi, email: hykychoi@cau.ac.kr Young Yang, email: yyang@sookmyung.ac.kr Keywords: suspension cells; metastasis; lipidomic profile; metabolic profile Received: March 29, 2017      Accepted: November 15, 2017      Published: December 07, 2017 ABSTRACT Circulating tumor cells (CTCs) are essential for the establishment of distant metastasis. Numerous studies have characterized CTCs as metastatic precursors; however, the molecular nature of CTCs has not been completely revealed yet due to the low number of CTCs in the blood stream. As an alternative approach, we developed a long-term suspension cell culture model using human breast cancer cell lines to mimic CTCs. We found that more than 40 passaged suspension cells acquired the ability to enhance metastasis like cancer stem cells. To identify molecular changes acquired during the suspension cell culture, we analyzed metabolic and lipidomic profiles as well as transcriptome in MDA-MB-468 suspension cells. Glutamate and leucine levels increased in suspension cells, and cholesterol synthesis pathway was altered. The inhibition of glutamate metabolic pathway decreased the proliferation of suspension cells compared to that of adherent cells. In the lipidomic profile, PC species containing long chain and polyunsaturated fatty acids increased in suspension cells and these species could be authentic and specific biomarkers for highly metastatic cancers. As this CTC-mimicking suspension cell culture model may easily apply to various types of cancer, we suggest this model as a great tool to develop therapeutic targets and drugs to eradicate metastatic cancer cells.