Citron Rho-Interacting Serine/Threonine Kinase Promotes HIF1a-CypA Signaling and Growth of Human Pancreatic Adenocarcinoma

Figure 5

CypA contributes to the function of CIT in regulating growth and apoptosis of PDAC cells. (a) CypA knockdown in Capan-1 cells. Capan-1 cells were infected with lentivirus carrying sh-Ctrl or sh-CypA for 24 hours, qPCR and western blot were used to analyze CypA expression. . (b) CypA knockdown induces apoptosis of Capan-1 cells. Capan-1 cells were infected with lentivirus carrying sh-Ctrl or sh-CIT for 48 hours, and then apoptosis of the cells was monitored. vs. sh-Ctrl. (c) CypA knockdown induces cell cycle arrest in Capan-1 cells. Capan-1 cells were infected with lentivirus carrying sh-Ctrl or sh-CIT for 48 hours, and then the cell cycle was monitored. , . (d) CypA knockdown blocks the effects of CIT on the proliferation of Capan-1 cells. Capan-1 with stable CIT overexpression, CypA knockdown, or both were subjected to cell proliferation experiment. vs. sh-Ctrl; , vs. sh-Ctrl. (e) CypA knockdown blocks the effects of CIT on colony formation of Capan-1 cells. Capan-1 with stable CIT overexpression, CypA knockdown, or both were subjected to colony formation assay, and the colonies were cultured for 14 days before final analysis. vs. sh-Ctrl; vs. sh-Ctrl. (f) CypA overexpression in Capan-1 cells. Capan-1 cells were infected with lentivirus overexpressing CypA or control construct for 24 hours. Then, CYPA expression was analyzed by western blot. (g) CypA overexpression rescues the effects of CIT on the proliferation of Capan-1 cells. Capan-1 with stable CIT knockdown, CypA overexpression, or both were subjected to cell proliferation experiment. vs. sh-Ctrl; , vs. sh-Ctrl. (h) Inhibition of CypA receptor CD147 blocks the effects of CIT on the proliferation of Capan-1 cells. Capan-1 with/without stably CIT overexpression was treated with CD147 inhibitor AC-73 (5 μM). vs. Ctrl + DMSO; , vs. Ctrl + DMSO.

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