| 研究実績の概要 |
In the preset study I investigate the relationship between JSAP-mediated lysosome trafficking and cell death under acute oxidative stress condition. For the FY 2019, we established the role of JLP (JSAP2) in curcumin-induced cell death and clarified the molecular mechanisms. In FY 2020, we expanded our observation to JSAP1, a MAPK scaffold protein which has a similar structure to JLP (JSAP2). Unexpectedly, we found that JSAP1 and JLP differentially regulate curcumin/ROS-induced cell death. We also found that JSAP1 might have different role in autophagy proses to JLP, although further analysis is required. All of our observations have been published in peer-reviewed journal, two in 2020 and one in 2021.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
1: 当初の計画以上に進展している
理由
We have established shRNA-mediated knockdown and Crispr/Cas9-mediated knockout of JSAP1 in HCT116 (human colorectal cancer cell line). The results indicated that JSAP1 and JLP, despite a high sequence homology, are differently regulate curcumin-induced cell death. Analysis on autophagic flux unexpectedly showed an accumulation of autolysosome, suggesting that JSAP1 regulates autophagy at much later step, unlike previously expected. In addition, we expand our observation using normal human cell line (RPE1-hTERT), by knockdown of JLP using siRNA. Unexpectedly, unlike observation in cancer cell line, we observed a decreased in cell proliferation, suggesting a link between JSAP-mediated transport and cell proliferation.
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| 今後の研究の推進方策 |
For future analysis, we plan to analyze how JSAP-mediated transport may related to the cell proliferation of normal cells. We plan to analyze on different normal cell (e.g. primary mouse embryonic fibroblast) to see the generality of the phenotype. To get insight to the molecular mechanism, we plan to analyze the possibility of cell cycle arrest at a specific phase by FACS, and then analyze the expression of key genes at that particular phase.
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