Daphnoretin-induced apoptosis in HeLa cells: a possible mitochondria-dependent pathway

Abstract

Daphnoretin is a bicoumarin compound isolated from a natural product, Wikstroemia indica, which has been used to treat many diseases. It has strong antiviral and anti-tumor activities. Taking the anti-tumor activity of daphnoretin as a starting point, the present study aimed to test the pro-apoptotic effect of daphnoretin and its underlying mechanism in HeLa cells. The inhibitory effects of daphnoretin on viability and proliferation of HeLa cells were determined by the MTT assay. Daphnoretin-induced apoptotic morphological changes were analyzed by mitochondrial membrane potential and Hoechst staining. The number and stage of apoptotic HeLa cells were determined by flow cytometry. Gene expression was determined by reverse-transcription polymerase chain reaction. Protein expression was determined by western blot. The caspase activity of HeLa cells was detected by a caspase-3 and caspase-9 colorimetric assay kit. We found that daphnoretin significantly inhibited HeLa cells' viability by the MTT assay and flow cytometry. The nuclei of the apoptotic cells exhibited strong, blue fluorescence in Hoechst staining. Bax mRNA and protein levels were increased while bcl-2 mRNA levels were decreased after daphnoretin treatment. Daphnoretin also activated both caspase-3 and caspase-9. These findings suggest that daphnoretin promotes apoptosis of HeLa cells in a mitochondria-mediated way. Daphnoretin therefore has potential to be a promising drug to treat uterine cervix cancer.

Effect of daphnoretin on the proliferation and apoptosis of A549 lung cancer cells in vitro

“…( 16 ) as well as Daphne mezereum L. and Daphne cannabina Wall. ( 17 ), have been shown to induce cell cycle arrest and apoptosis in leukemia, osteosarcoma and HeLa cells ( 13 – 15 ). These findings suggest that daphnoretin may have potential as a novel anti-cancer agent.…”

“…Previous studies have shown that daphnoretin has a number of biological activities, including antifungal effects ( 8 ), as well as the inhibition of various sites involved in DNA synthesis ( 9 ), the activation of protein kinase C in platelet aggregation ( 7 , 10 ), an antiviral effect on hepatitis B ( 11 ) and respiratory syncytial virus properties ( 12 ). Furthermore, the anticancer effect of daphnoretin has been reported in various studies ( 13 – 15 ). Such studies have shown that daphnoretin exerts its anticancer effects through the inhibition of cancer cell proliferation, the induction of G 2 /M-phase arrest and apoptosis.…”

Daphnoretin induces reactive oxygen species‑mediated apoptosis in melanoma cells

“…A study demonstrated that daphnoretin induced apoptosis in human osteosarcoma cells by triggering G 2 /M cell cycle arrest and activating the caspase-3 pathway (21). In another study, daphnoretin was shown to induce the mitochondria-mediated apoptosis of HeLa cells (22). Daphnoretin has also been reported to influence the proliferation and apoptosis of A549 lung cancer cells in vitro (23).…”

Rapid detection of low‐level HeLa cell contamination in cell culture using nested PCR

“…Unlike other cell lines, one of the characteristics of HeLa cells is their abnormally rapid proliferation rate. Hela cells can adapt to different growth conditions and different cell culture media, such as DMEM, MEM, RMPI1640, DMEM/F12K, and are very easy to culture. Therefore, HeLa cells are one of the most important sources of cell cross‐contamination.…”