| 研究実績の概要 |
In this project, the organoids with different combination of driver gene mutations were simulated with activin and the cell survival and morphological changes were examined.
Cell survival: after activin treatment, the organoids with Kras mutation could survive well, otherwise, the organoids without Kras mutation go through apoptosis. Furthermore, blocking Kras-MEK signaling pathway by MEK inhibitors, the organoids with Kras mutation go through apoptosis, which suggests that Kras mutation inhibits activin-induced cell apoptosis.
Morphological changes: after activin treatment, the morphological changes were examined. The organoids with p53 homozygous mutation instead of heterozygous mutation form EMT-like morphology. Furthermore, overexpression of wt-p53 in p53 homozygous mutated organoids inhibits activin-induced morphological changes, which suggests that p53 homozygous mutation interacts with activin to induce morphological changes.
In vivo: to confirm activin-induced morphological changes associated with metastasis, we pretreated the organoids and injected into tail vein of the mice. By examining lung metastasis, we found that activin treated organoids form more metastasis.
Mechanism: to clarify the mechanism by which p53 interacts with activin to induce morphological changes, we performed RNA-seq and found that after activin simulation, some signaling pathways, such as Wnt, p38-MAPK, Stat3 signaling, were significantly upregulated. According to these findings we will test some inhibitors for inhibiting metastasis and the candidate inhibitors may be used for clinical trial.
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