Induction of apoptosis by casticin in cervical cancer cells through reactive oxygen species-mediated mitochondrial signaling pathways

Abstract

Abstract. Casticin, one of the main components from Fructus Viticis, has been reported to inhibit the growth of various cancer cells, including the human cervical cancer cell line HeLa. The purpose of this study was to examine the apoptotic activity and molecular mechanism of casticin action on human cervical cancer cells. The apoptotic activity of casticin on human cervical cancer HeLa, CasKi, SiHa and peripheral blood mononuclear cells (PBMCs) was measured using a histone/DNA ELISA assay, flow cytometry with propidium iodide (PI) staining and DNA agarose gel electrophoresis. The mitochondrial membrane potential and reactive oxygen species (ROS) production were evaluated by flow cytometry analysis. Caspase activities were assayed using a caspase colorimetric activity assay kit. Protein expression levels of cytochrome c, Bax, Bcl-2, Bcl-xL and XIAP were analyzed by Western blotting. Casticin caused accumulation of the Sub-G1 cells and increased reactive oxygen species (ROS) production in HeLa, CasKi, SiHa cell lines, but not in PBMCs. Apoptosis of HeLa cells was induced by casticin via mitochondrial release of cytochrome c due to the reduction of mitochondrial trans membrane potential, activation of caspase-3 and -9, and the production of reactive oxygen species. The pan caspase inhibitor zVAD-FMK, the caspase-9 inhibitor zLEHDfmk and N-acetylcysteine suppressed casticin-induced apoptosis. Bax was upregulated, while expression levels of Bcl-xL and XIAP were downregulated. However, there was no change in the expression of Bcl-2 under the same treatment. Our results indicate that casticin-induced apoptosis of cervical cancer cells is mediated by ROS generation and mitochondrial signaling pathways.

An Investigation into the Cytotoxic Effects of 13-Acetoxysarcocrassolide from the Soft Coral Sarcophyton crassocaule on Bladder Cancer Cells

“…However, previous studies also showed that the apoptosis-induced activity exerted by the natural plant extract or chemical compound is correlated with caspase-3 and caspase-9, but not caspase-8 [49,51,52] In the current study, the increase in cleaved caspase-3 and cleaved caspase-9 was verified by western blot compared with the control as cleaved caspase-8 was not found in BFTC cells after exposure to 13-acetoxysarcocrassolide. These results support the apoptosis-induced effects of 13-acetoxysarcocrassolide on BFTC cells displayed by flow cytometry and are in accordance with the evidence on caspase-3 and caspase-9-associated apoptosis shown previously [48,49,50,51,52]. Moreover, these data demonstrate that activation of caspase-3 and caspase-9 is likely involved in induction of apoptosis by 13-acetoxysarcocrassolide in BFTC cells.…”

“…Another member of the caspase family, caspase-9, has also been shown to be associated with the induction of apoptosis in several cancer cells including cervical cancer cells, human hepatoma cells, melanoma cells, and human lung cancer cells. Interestingly, the activation of caspase-3 is tightly linked to the activation of caspase-9 in these anti-tumor studies [48,49,50,51,52]. However, previous studies also showed that the apoptosis-induced activity exerted by the natural plant extract or chemical compound is correlated with caspase-3 and caspase-9, but not caspase-8 [49,51,52] In the current study, the increase in cleaved caspase-3 and cleaved caspase-9 was verified by western blot compared with the control as cleaved caspase-8 was not found in BFTC cells after exposure to 13-acetoxysarcocrassolide.…”

Polygonum cuspidatumExtract Induces Anoikis in Hepatocarcinoma Cells Associated with Generation of Reactive Oxygen Species and Downregulation of Focal Adhesion Kinase

“…Many natural products, such as casticin, curcumin, wogonin, Tanshinone IIA, and berberine, cause apoptosis through the mediation of ROS [25–29]. Emodin and curcumin may sensitize cancer cell to anoikis through ROS generation [14, 22].…”

Casticin induces breast cancer cell apoptosis by inhibiting the expression of forkhead box protein M1

“…Inhibition of FOXO3a by siRNA predominantly blocks casticin-induced apoptosis. Previous studies have demonstrated the antiproliferative and pro-apoptotic effects of casticin in prostate (10), cervical (14,15), lung (11) and colon (12) cancer. This study investigated the role of FOXO transcription factors in mediating the effects of casticin.…”

“…Casticin has also been reported to induce cell death of leukemia cells through the induction of apoptosis or mitotic catastrophe (13). We recently reported casticin-induced apoptosis of cervical cancer (14,15) and hepatocellular carcinoma (16) cells; however, the underlying mechanisms remain unclear.…”