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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: GSTZ1 deficiency promotes hepatocellular carcinoma proliferation via activation of the KEAP1/NRF2 pathway

Fig. 4

GSTZ1 negatively regulates the KEAP1/NRF2 pathway in human HCC and in liver tissue of mice. ac ARE promoter activities in GSTZ1-overexpressing Huh7 cells (left) and GSTZ1-KO HepG2 cells (right). a ARE promoter activities. The NRF2 activator tBHQ (100 μM for 6 h) and the antioxidant NAC (4 mM for 24 h) were used as positive and negative controls, respectively. b Relative mRNA expression of NRF2 target genes in GSTZ1-OE and GSTZ1-KO HCC cells, determined via qRT-PCR. c Western blotting to assess expression levels of NQO1, the NRF2 target, in total cell extracts, and of NRF2 in cytoplasmic and nuclear cell extracts, indicating the effect of GSTZ1 on NRF2 intracellular localization and transcription. β-actin, β-tubulin, and Lamin B1 served as reference proteins for total, cytoplasmic, and nuclear extracts, respectively. d-f Association between GSTZ1 and NQO1 expression in HCC tissues. d Linear regression analysis showing the negative correlation between GSTZ1 and NQO1 mRNA expression in 40 cases of HCC (p = 0.0197; r = − 0.37, Pearson correlation coefficient). e Representative immunohistochemistry images for GSTZ1 and corresponding NQO1 in HCC tissues and tumor-adjacent normal tissues. f Western blotting for GSTZ1 and NQO1 in 9 pairs of HCC and tumor-adjacent normal tissues. For Western blotting, 50 μg protein was loaded per well. Values represent the mean ± SD (n = 3, performed in triplicate), *p < 0.05, **p < 0.01, Student’s t-test. ARE, antioxidant response element; HCC, hepatocellular carcinoma; qRT-PCR, quantitative reverse transcription polymerase chain reaction

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