Bcl-2/BCL2L12 mediated apoptosis and cell cycle arrest induced by Kaempferol through the suppression of PI3K/AKT signaling pathway in Hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a highly aggressive and third leading cancer-related death. PI3k/AKT and p53 signaling pathways play a vital role and regulate cellular proliferation, migration, cell cycle, apoptosis, and autophagy during cancer conditions. BCL2L12 is a bcl-2 family protein and encoded by the BCL2L12 gene, which is involved in the execution phase of apoptosis.  However, the biological function of PI3K/AKT/Bcl-2 mediated BCL2L12, and its molecular mechanism in HCC is largely unknown. Herein, we investigated the effect of Kaempferol, on PI3K/AKT/Bcl-2/BCL2L12 expressions in HCC cells. The expression level of PI3K/AKT/Bcl-2/BCL2L12, and its target genes in polyphenolic flavonoid Kaempferol treated HCC cells analyzed by Quantitative Real-Time PCR and Western blotting. Subsequently, the effect of Kaempferol in HCC chemosensitivity, cell proliferation, migration, cell cycle, and apoptosis were analyzed using AO/EtBr staining, ROS staining, mitochondrial membrane potential assay, wound healing assay, Transwell migration assay, MTT assay, and PI Staining. We found that the downregulation of PI3-K, AKT, LC3A/B, MMP9, Bcl-2, and BCL2L12 and upregulation of PTEN, p53, p21, and caspase 3 in Kaempferol treated HCC cells. Subsequently, we observed that Kaempferol effectively inhibits cellular proliferation, migration and enhances apoptosis, cell cycle arrest, autophagy, and 5-Fluorouracil mediated chemosensitivity in HCC cells. Furthermore, Kaempferol has dominant role against HCC and inhibits 50-60% of cells after 24h treatment with 30μM in HepG2 and 40μM in Huh7 cells. Therefore, Kaempferol plays a promising role in cancer therapeutics by inhibiting PI3K/AKT/Bcl-2 mediated BCL2L12 expression in HCC.