Fig. 2

Transiently induced expression of hMT3 increases the resistance of intrinsically susceptible Huh7 cells to sorafenib. A Analysis of a baseline expression of hMT3 in Huh7^WT and BCLC-3^WT cells, accompanied by FR value obtained by cDNA microarray. *p ≤ 0.05, **p ≤ 0.01 (Student’s test, two-sided). B Representative micrographs of Huh7^WT cells and the same cells transfected either with pcDNA3.1-GFP-TOPO (Huh7^mock transfection) or pcDNA3.1-GFP-hMT3-TOPO (Huh7^hMT3). Scale bar, 10 μm. C Effect of hMT3 up-regulation on proliferation rate of Huh7 cells analyzed using real-time electrical impedance measurement. D Dose-response metabolic activity of Huh7^mock and Huh7^hMT3 cells exposed to sorafenib (24 h). Data are expressed as mean ± SEM (n = 3). *p ≤ 0.05, **p ≤ 0.005, ***p ≤ 0.0001. E Representative micrographs of spheroids formed by Huh7^mock and Huh7^hMT3 cells exposed to sorafenib (24 h). Scale bars, 200 μm. F MT3 expression in normal liver (n = 50) and HCC (n = 50) tissues. G MT3 expression in various stages of HCC (A, early stage; B, intermediate stage; C, advanced stage) in patients either treated (red) or untreated (blue) with sorafenib. The data were gathered and analyzed using the cBioPortal for Cancer Genomics. H The Kaplan-Meier curves indicating that in the studied HCC cohort (n = 364, RNA-seqID: 4504), subjects with high expression of MT3 exhibited worse prognosis than patients with low expression of MT3. (I) The Kaplan-Meier curves showing that HCC patients with high expression of MT3 treated with sorafenib (n = 30) exhibited significantly lower survivability than sorafenib-treated subjects with low MT3 expression. Data were derived from Kaplan-Meier plotter database. p ≤ 0.05 was considered statistically significant