Proteomic Investigation into Betulinic Acid-Induced Apoptosis of Human Cervical Cancer HeLa Cells

Abstract

Betulinic acid is a pentacyclic triterpenoid that exhibits anticancer functions in human cancer cells. This study provides evidence that betulinic acid is highly effective against the human cervical cancer cell line HeLa by inducing dose- and time-dependent apoptosis. The apoptotic process was further investigated using a proteomics approach to reveal protein expression changes in HeLa cells following betulinic acid treatment. Proteomic analysis revealed that there were six up- and thirty down-regulated proteins in betulinic acid-induced HeLa cells, and these proteins were then subjected to functional pathway analysis using multiple analysis software. UDP-glucose 6-dehydrogenase, 6-phosphogluconate dehydrogenase decarboxylating, chain A Horf6-a novel human peroxidase enzyme that involved in redox process, was found to be down-regulated during the apoptosis process of the oxidative stress response pathway. Consistent with our results at the protein level, an increase in intracellular reactive oxygen species was observed in betulinic acid-treated cells. The proteins glucose-regulated protein and cargo-selection protein TIP47, which are involved in the endoplasmic reticulum pathway, were up-regulated by betulinic acid treatment. Meanwhile, 14-3-3 family proteins, including 14-3-3β and 14-3-3ε, were down-regulated in response to betulinic acid treatment, which is consistent with the decrease in expression of the target genes 14-3-3β and 14-3-3ε. Furthermore, it was found that the antiapoptotic bcl-2 gene was down-regulated while the proapoptotic bax gene was up-regulated after betulinic acid treatment in HeLa cells. These results suggest that betulinic acid induces apoptosis of HeLa cells by triggering both the endoplasmic reticulum pathway and the ROS-mediated mitochondrial pathway.

Molecular mechanisms responsible for programmed cell death-inducing attributes of terpenes from Mesua ferrea stem bark towards human colorectal carcinoma HCT 116 cells

“…Results of present study are consistent with other research reports where selective increase in ROS generation in cancer cells was observed after treatment with natural products. Similarly, 5-Fluorouracil, a standard drug to treat CRC, has also been reported to induce cell death through generation of ROS (Fu et al, 2014;Wen et al, 2002;Xu et al, 2014;Zou et al, 2015). After establishing the ROS promoting role of F-3, we further confirmed the results by a series of in vitro antioxidant assays i.e., DPPH, ABTS and FRAP, and findings of the study revealed that F-3 phytoconstituents has weak activity in all the models employed.…”

Betulinic Acid: Recent Advances in Chemical Modifications, Effective Delivery, and Molecular Mechanisms of a Promising Anticancer Therapy

“…The antiproliferative and apoptosis‐inducing mechanisms of BetA explain its mode of action. Biological activity of BetA was first demonstrated in melanoma cell lines and later in breast , brain tumor , prostate , colon, leukemia , head and neck , and cervical tissues (Table ). Activity has been shown in primary tumor samples and later confirmed in melanoma xenograft mouse models .…”

“…Besides the above mechanisms of action, BetA also acts in a cell-specific manner (88,(122)(123)(124). It is well known that NF-jB plays a major role in stress-induced transcriptional activation.…”

Knockdown of Sucla2 decreases the viability of mouse spermatocytes by inducing apoptosis through injury of the mitochondrial function of cells

“…ROS detection kit (s0033, Beyotime Biotech, Shanghai, China) and flow cytometry were used to detect the reactive oxygen species (ROS) in the cells, as described in a previous paper [14]. 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA, Sigma-Aldrich, Saint Louis, MO, USA) is a fluorescent dye with no fluorescence.…”