title: Molecular changes in appearance of a cancer cell among normal HEK293T cells.
abstract: In very early stages of cancer development, one or a few cells expressing cancer-associated genes appear among a much larger number of surrounding normal cells. To analyze the molecular changes induced by this co-existence, we artificially prepared transformed cells with complete loss of tumor suppressor gene, SCRIB, among normal human embryonic kidney (HEK293T) cells. A cell strain with SCRIB-knockout was successfully constructed by using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 nuclease system and co-cultured with normal cells. By measuring the time-course changes in cell numbers when SCIRB-knockout cells (cancer model) or cells with normal level of SCRIB expression were respectively co-cultured with wild-type normal HEK293T cells, it was shown that the SCRIB-knockout strain was beneficial for proliferation when mixed together with normal cells. Moreover, as a result of proteome analysis on wild-type cells separated from co-culture with SCRIB-knockout cells, a total of 843 proteins were identified, among which 139 proteins were specific. Among the specifically identified proteins, 22 proteins were annotated to be involved in cytoskeletons including microtubule motor activity-associated proteins. It was implied that molecular changes in cytoskeletons occurred in normal cells when co-cultured with SCRIB knockout cells, but the SCRIB knockout might affect proliferation of the transformed cells with SCRIB knockout by defensive or offensive mechanism of surrounding normal cells.
authors: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
authors: Jihye
authors: J
authors: Chung
authors: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan; Japan Society for the Promotion of Science, Sakyo-ku, Kyoto 606-8502, Japan.
authors: Shunsuke
authors: S
authors: Aburaya
authors: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
authors: Wataru
authors: W
authors: Aoki
authors: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: miueda@kais.kyoto-u.ac.jp.
authors: Mitsuyoshi
authors: M
authors: Ueda
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics; D049109:Cell Proliferation; D018920:Coculture Techniques; D003599:Cytoskeleton / Q000378:metabolism; D000072669:Gene Editing; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D020543:Proteome / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000172:deficiency / Q000235:genetics
chemical_list: D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D020543:Proteome; C480591:SCRIB protein, human; D025521:Tumor Suppressor Proteins
keywords: Cellular competition; Co-culture; HEK293T; Proteome analysis; Tumor suppressor gene