Fig 2: Distinct interaction between the testosterone tetrad elicits androgenic signals that drive cell cycle progression, enhanced motility, cancer stemness, and resistance to therapy in patients with PCa. STRINGdb-generated visualization of the protein–protein interaction between (A) HSD17B2, HSD17B3, SHBG, and SRD5A1, as well as with (B) SRD5A2, SRD5A3, HSD3B1, HSD3B2, and CYP17A1. (C) Molecular docking showing the direct interaction between HSD17B2 and HSD17B3, SRD5A1 and SHBG (upper left), as well as protein tetrad formation by the HSD17B2/HSD17B3 and SRD5A1/SHBG complexes (lower right). Heatmaps showing the correlation between expression profile of the testosterone tetrad and molecular effectors of (D) multidrug resistance, and (E) cancer stemness, in the GSE26910 mixed prostate and breast cancer cohort. Columns with similar annotations are collapsed by taking mean inside each group. Rows are centered; unit variance scaling is applied to rows. Both rows and columns are clustered using correlation distance and average linkage. (F) Heatmap showing the correlation between expression profile of the testosterone tetrad and ABCC1, ABCB1, ABCG2, ALDH1A1 expression in GSE33455 docetaxel-resistant PCa cell lines dataset. (G) Transcriptional profiling heatmap showing the correlation between the differential expression of the testosterone tetrad in human benign prostatic hyperplasia, primary and metastatic PCa samples from the GSE3325 PCa cohort. (H) Molecular docking showing the interaction cascade between the testosterone tetrad and molecular effectors or mediators of cell cycle progression, and drug uptake/therapy response.

Fig 3: Expression of ATP-binding cassette (ABC) transporters in Calu-3 cells and EpiArway™ cultured under ALI conditions for 8 d or 21 d. Left: the mRNAs for the genes of interest were determined by means of RT-qPCR analysis and normalized for that of the reference gene (RPL15), according to the formula described in Methods. Data are means ± SEM of three independent experiments. ** p < 0.01, *** p < 0.001 vs. Calu-3 8 d; $$ p < 0.01, $$$ p < 0.001 vs. Calu-3 21 d; n.d. not detectable. Right: the expression of the transporters at the protein level was addressed by means of Western blot analysis, as described in Methods. A representative blot is shown, repeated three times with comparable results. MDR1: P-glycoprotein; MRP1: multidrug resistance-associated protein 1; BCRP: breast cancer resistance protein.

Fig 4: Effects of different theaflavin monomers on the mRNA and protein levels of MRP1, MRP2, MRP3, P-gp and BCRP in Caco-2 cells. (A) Effect of theaflavins on mRNA levels of efflux transporters. (B) Effect of theaflavins on protein levels of efflux transporters. Ctrl: Caco-2 cells were incubated with HBSS; TF, TF3G, TF3’G and TFDG: Caco-2 cells were incubated with TF, TF3G, TF3’G and TFDG. Results are expressed as means ± SD (N = 3). Different small letters indicate significant difference between different treatments at p<.05.

Fig 5: DDP-resistant ovarian cancer cells possess enhanced metastatic activity and arachidonic acid metabolism. a The procedure for the induction of DDP-resistant SKOV3 cells (upper panel); IC50 and the cell viabilities of SKOV3 and SKOV3-R with different concentrations of DDP treatment (lower panel). b, c The mRNA (b) and protein (c) levels of MRP1, MRP4, Slug and Snail in SKOV3 and SKOV3-R. d Liver metastasis in nude mice after SKOV3 or SKOV3-R subcutaneous transplantation. e, f. Migration and invasion of SKOV3 and SKOV3-R were tested by wound-healing (e) and transwell assays (f). g MRM chromatograms of AA, TXB2, 12-HETE, 5,6-EET, and 14,15-EET. h The supernatant levels of LTB4, 5-HETE, 12-HETE, PGE2, TXB2, 5,6-EET, 14,15-EET, 14,15-DHET, 11-HETE, and 11,12-DHET in SKOV3 and SKOV3-R. i The heatmap for gene expression of 12LOX, COX1, CYP2J2, COX2, 5LOX, 15LOX, LTA4H and PGDH. j the protein levels of 12LOX and COX1 were assessed by western blotting. Data are expressed as mean ± SEM of 4 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001