Manilkara zapota (L.) P. Royen Leaf Water Extract Induces Apoptosis in Human Hepatocellular Carcinoma (HepG2) Cells via ERK1/2/Akt1/JNK1 Signaling Pathways

Figure 9

Expression of ERK1/2, Akt1, JNK1, iNOS, and VEGFA at mRNA levels in HepG2 cells incubated with Manilkara zapota leaf water extract and 5-Fluorouracil (5-FU) for 72 h analyzed using quantitative real-time PCR. Values are reported as mean ± SD (n = 3). Value with different superscript letter indicates significant difference between groups by Tukey’s test (P < 0.05). The mRNA expression of ERK1/2 was significantly downregulated at 48 and 96 μg/mL of Manilkara zapota leaf water extract treated HepG2 cells (P < 0.05). Manilkara zapota leaf water extract significantly upregulated the ERK1/2 mRNA level at 24 μg/mL (P < 0.05). Akt1 expression was significantly downregulated in HepG2 cells treated with 24, 48, and 96 μg/mL of Manilkara zapota leaf water extract compared to the control (P < 0.05). Treatment with Manilkara zapota leaf water extract in a concentration of 48 and 96 μg/mL significantly upregulated the JNK1 mRNA level (P < 0.05). Treatment with 24 μg/mL of Manilkara zapota leaf water extract significantly upregulated the gene expression of iNOS (P < 0.05). Manilkara zapota leaf water extract significantly reduced the gene expression of VEGFA compared to the untreated cells (P < 0.05). ACTB: beta-actin; ERK1/2: extracellular signal-regulated kinase 1/2; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; iNOS: inducible nitric oxide synthase; JNK1: c-Jun N-terminal kinase 1; VEGFA: vascular endothelial growth factor A.