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

Figure 4

CIT promotes the expression of CypA in a HIF1-dependent manner. (a) CIT expression is correlated with CypA in PDAC tissues. The expressions of CIT and CypA in PDAC tissues were analyzed using the data from the TGCA database with the web tool GEPIA (http://gepia.cancer-pku.cn). (b) CIT knockdown reduces the protein level of CypA. Capan-1 and BxPC-3 cells were infected with lentivirus carrying sh-Ctrl or sh-CIT for 24 hours, and then western blot was applied to analyze the expression of CypA. (c) CIT knockdown reduces the mRNA level of CypA. Capan-1 and BxPC-3 cells were infected with lentivirus carrying sh-Ctrl or sh-CIT for 24 hours, then qPCR was applied to analyze the expression of CypA. vs. sh-Ctrl. (d) CIT overexpression increases the level of CypA. Capan-1 cells were infected with lentivirus carrying Ctrl or CIT expressing construct for 24 hours, then qPCR and western blot were applied to analyze the expression of CypA. vs. Ctrl. (e) HIF-1 inhibition blocks the effects of CIT on CypA expression. Capan-1 cells were infected with lentivirus carrying Ctrl or CIT expressing construct for 24 hours in the presence of HIF-1 inhibitors BAY 87-2243 (1 μM), LW 6 (10 μM), or DMSO, and then qPCR was applied to analyze the expression of CypA. vs. Ctrl + DMSO. (f) CIT overexpression increases the protein level of HIF1A. Capan-1 cells were infected with lentivirus carrying Ctrl or CIT expressing construct for 24 hours, then qPCR and western blot were applied to analyze the expression of HIF1A.

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