Combining the chemotherapeutic effects of epigallocatechin 3-gallate with siRNA-mediated p53 knock-down results in synergic pro-apoptotic effects

Figures & data

Figure 1 Cytotoxic effects as measured by MTT assay after 24 hours incubation with 50 nM p53siRNA, in the presence or absence of 10 μM EGCG, and siPORT™ NeoFX™ transfection agent on HeLa cell line (mean ± standard deviation, n = 6; P ≤ 0.05 compared with the control group).

Abbreviations: EGCG, (−)-epigallocatechin-3-gallate; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; siRNA, short interfering RNA.

Table 1 Apoptosis-related genes with altered expression levels following 24 hours of incubation with p53siRNA, p53siRNA+EGCG, or EGCG

Gene family	p53siRNA	p53siRNA+EGCG	EGCG
TNF ligand family	TNFSF10 (+1.47)	FASLG (+1.29), TNFSF10 (+9.05), TNF (−1.54), TNFRSF10A (−2.31), TNFRSF10B (−2.56), TNFRSF25 (−2.80), CD70 (−1.48)	TNFRSF10 (+2.3), TNFRSF11B (−1.27)
TNF receptor family	TNFRSF11B (−1,43), CD27 (−1.72)	CD27 (+3.87), CD40 (−1.67), LTBR (−1.68)	LTBR (+1.307), TNFRSF21 (+1.5), CD27 (+1.54)
Bcl-2 family	BCL2L1 (−1.47), BCL2L2 (−1.27), MCL1 (−1.46), NOL3 (−1.48)	BCL2A1 (+4.08), BAD (−1.61), BAX (−2.06), BCL2L1 (−2.06), BCL2L2 (−2.53), MCL1 (−1.89), NOL3 (−1.89)	BAK1 (+1.27), BNIP1 (−1.25)
Caspase family	CASP1 (+2.66)	CASP4 (+1.28), BIRC2 (−1–33), BIRC3 (−1.34), CASP2 (−1.56)	BIRC2 (−1.33)
TRAF family	TRAF2 (−1.5)	TRAF2 (−2.98), TRAF3 (−1.52), TRAF4 (−1.96)
CARD family	CARD8 (−1.41)	CRADD (+1.55), PYCARD (+1.29), RIPK2 (+1.25), APAF1 (−1.31), CASP2 (−1.56)	CARD8 (−1.39)
Death domain family		FADD (−1.73), TNFRSF10A (−2.31), TNFRSF10A (−2.31), TNFRSF10B (−2.56), TNFRSF25 (−2.80)	TNFRSF21 (+1.5), TNFRSF11B (−1.27)
Death effector domain family	CFLAR (−1.29), TNFRSF11B (−1.43)	CFLAR (−1.5)	CFLAR (−1.34)
CIDE domain family		CIDEB (2.94), DFFA (1.54)
p53 and DNA damage response	TP53 (−1.73), BCL2L1 (−1.47)	TP53 (−2.06), ABL1 (−1.53),	TP73 (+1.81)
Anti-apoptosis	BCL2L2 (−1.27)	BRAF (+2.11), AKT1 (−1.31)

Abbreviations: CARD, caspase recruitment domain; EGCG, (−)-epigallocatechin-3-gallate; siRNA, short interfering RNA; TNF, tumor necrosis factor; TRAF, TNF receptor-associated factor.

Figure 2 The clustergram generated for differentially expressed genes (P < 0.05, cut-off fold change 1.25) in treated versus control samples.

Figure S1 The canonical network associated with the genes involved in cancer apoptosis mediated by (A) EGCG, (B) p53siRNA, and (C) p53+EGCG.

Figure S2 The canonical network associated with the genes involved in death receptor signalling mediated by (A) EGCG, (B) p53siRNA, and (C) p53+EGCG.