Figure 8.

Met inhibits the expression of CAIX in HTB-35 cells under hypoxic conditions. The cells were incubated either in normoxia (21% O₂ level) or in hypoxia (5% O₂ level) with CA, Met or both compounds for 24 h. qPCR analysis was performed to assess the expression of HIF-1α under normoxic and hypoxic conditions (A). The RNA expression of HIF1A was determined with the 2^−ΔΔCt method. The mRNA level of HIF1A was significantly increased following hypoxia ((A), p < 0.01 vs. control). On the contrary, Met alone and together with CA downregulated HIF1A ((B), p < 0.01 vs. control for Met, p < 0.01 vs. control for CA/Met). RT-PCR analysis revealed that in hypoxia the expression of CAIX significantly decreased in cells exposed to Met and CA/Met ((C), HPRT1 was a reference gene). Data shown here are representative of three experiments performed with similar results (C).