Fig. 6: RPRD1A induced ROS inhibition correlates with chemotherapeutic resistance.

A Cell survival of MHCCLM3 NC and shRPRD1A cells exposed to serial dilutions of OXP for 60 h. B Western blot of lysates generated from MHCCLM3-NC and -shRPRD1A cells treated with OXP as indicated. C ROS levels were detected by flow cytometry in MHCCLM3 NC and shRPRD1A cells treated with 50 μM OXP or control for 4 h. D Quantification of the ROS levels in (C). E Cell survival of control and MHCCLM3-RPRD1A cells exposed to serial dilutions of oxaliplatin for 60 h. F Western blotting of lysates generated from control and MHCCLM3-RPRD1A cells treated with OXP as indicated. G, H Caspase 3/7 assay was performed to detect the activity apoptosis of MHCCLM3-NC and MHCCLM3-shRPRD1A (G), control and MHCCLM3-RPRD1A (H) treated with 50 μM OXP for 24 h. I Annexin V and PI staining analysis were performed to test cell apoptosis of MHCCLM3-NC, MHCCLM3-shRPRD1A and RPRD1A supplemental cells treated with or without 100 μM OXP for 6 h. J Histogram was the quantification. K Representative images of RPRD1A and NRF2 staining in HCC specimens. Group 1, low NRF2 and low RPRD1A expression (n = 64); group 2, high NRF2 and low RPRD1A expression (n = 110); group 3, low NRF2 and high RPRD1A expression (n = 22); and group 4, high NRF2 and high RPRD1A expression (n = 71). The cross table showed the distribution of RPRD1A and NRF2 expression in HCC samples (n = 267); the Pearson λ² correlation coefficient and p value are shown. L, M Kaplan–Meier analysis of OS (L) and DFS (M) in HCC patients according to RPRD1A and NRF2 expression. N, O Kaplan–Meier analysis of OS (N) and DFS (O) in HCC patients with TACE according to RPRD1A expression. In panels (D), (H), (J), the p values were determined by a two-tailed t-test. In panels L–O, the p values were determined by the log-rank test. *p < 0.05, **p < 0.01, ***p < 0.001. Results are representative of at least three independent experiments.