Beta-Amyrin Modulates P38 MAPK and Jnk Pathway to Inhibit Cell Proliferation and Induce ROS-mediated Apoptosis in HeLa Cells

Abstract

Anburaj et al.: β-Amyrin induces ROS-mediated apoptosisIt is aimed to investigate the effect of β-amyrin on p38 mitogen-activated protein kinase and Jun N-terminal kinase pathways and apoptosis in HeLa cells. HeLa treated cells were divided into 6 groups, group I-HeLa untreated cells as control, group II-dimethyl sulfoxide serve as vehicle control, group III-cisplatin as standard drug, group IV-β-amyrin-treated HeLa cells, group V-cells were pretreated with 100 μm N-acetyl-L-cystein for 1 hour and then treated with cisplatin and group VI-cells were pretreated with 100 μm N-acetyl-L-cystein for 1 hour and then treated with β-amyrin. The antiproliferative effect was measured using the MTT assay. Genotoxic effects were studied using micronucleus assay. Total reactive oxygen species, nitric oxide and caspase 3 level were determined on a spectrofluorimeter and colorimeter. Protein expression was analyzed by immunoblotting. β-Amyrin (10-200 μm) and cisplatin (0.01-100 μm) had an inhibitory effect on the proliferation of cancer cells in a dose-dependent manner, with the IC50 values at 100 μm and 10 μm for β-amyrin and cisplatin, respectively. Western blot analysis revealed expressions of apoptotic pathway related proteins, Bcl-2, caspase-3, caspase-9, phospho-p38 mitogen-activated protein kinase and phospho-Jun N-terminal kinase, growth arrest and deoxyribonucleic acid-damage-inducible, beta in all groups. Genotoxic effects were observed after treatment with β-amyrin as well as with cisplatin. It was observed that HeLa cells showed significant elevation of total reactive oxygen species after β-amyrin treatment. Protein expression analysis showed that the β-amyrin upregulated phospho-p38 mitogenactivated protein kinase, phospho-Jun N-terminal kinase and growth arrest and deoxyribonucleic aciddamage-inducible, beta on HeLa cells. Increased phospho-Jun N-terminal kinase directly activated caspases and decreased Bcl-2 in HeLa cells. These results indicated that β-amyrin induced the apoptosis through reactive oxygen species-mediated mechanism by activating p38 mitogen-activated protein kinase and Jun N-terminal kinase through transcriptional factor, GADD45β. In turn, activated Jun N-terminal kinase directly activated caspase-9 and caspase-3 and destined the HeLa cells to apoptosis.

Antimicrobial and Cytotoxic Activities of Selected Hieracium L. s. str. (Asteraceae) Extracts and Isolated Sesquiterpene Lactones

“…The most active was H. scheppigianum CH 2 Cl 2 extract, which effect was the strongest on HeLa cells ( IC 50 =148.1 μg/mL), followed by A549 and LS174 cells ( IC 50 =294.0 and 308.9 μg/mL). Cytotoxic effect of this extract could be at least partly explained by the presence of α ‐ and β ‐amyrins, which ability to inhibit HeLa and/or A549 cells growth was demonstrated previously (both compounds were active against HeLa cells IC 50 =29.4 and 42.7 μg/mL; α ‐amyrin also against A549 cells IC 50 =31.3 μg/mL) [38,39] . It has been also found that β ‐amyrin achieved this effect in HeLa cells by producing ROS and consequently activating the signaling pathways that led to apoptosis [39] …”

“…Cytotoxic effect of this extract could be at least partly explained by the presence of αand β-amyrins, which ability to inhibit HeLa and/or A549 cells growth was demonstrated previously (both compounds were active against HeLa cells IC 50 = 29.4 and 42.7 μg/mL; αamyrin also against A549 cells IC 50 = 31.3 μg/mL). [38,39] It has been also found that β-amyrin achieved this effect in HeLa cells by producing ROS and consequently activating the signaling pathways that led to apoptosis. [39] MeOH extracts also showed the highest activity on HeLa cells (IC 50 = 152.3 -303.2 μg/mL), followed by LS174 cells (IC 50 = 512.9 -689.7 μg/mL) and A549 cells (IC 50 = 625.2 -869.6 μg/mL).…”

“…[38,39] It has been also found that β-amyrin achieved this effect in HeLa cells by producing ROS and consequently activating the signaling pathways that led to apoptosis. [39] MeOH extracts also showed the highest activity on HeLa cells (IC 50 = 152.3 -303.2 μg/mL), followed by LS174 cells (IC 50 = 512.9 -689.7 μg/mL) and A549 cells (IC 50 = 625.2 -869.6 μg/mL). Like in the case of antimicrobial activity, the strongest effect was shown by H. coloriscapum, H. calophyllum and H. valdepilosum MeOH extracts, which did not contain the highest amounts of total polyphenols, however, all were rich in luteolin 7-O-glucoside.…”

A Network Pharmacology Approach to Explore Clinacanthus Nutans on Colorectal Cancer

“…β-amyrin was the most potent chemical identified from CN, according to the network. In a study of cytotoxicity in human cervical adenocarcinoma (HeLa) cell-line, βamyrin induced apoptosis via an increase in reactive oxygen species (ROS) (21). In a study by Maiyo and colleagues, when β-amyrin extracted from Prunus africana was tested on colorectal carcinoma (Caco-2) cell lines, a significant cytotoxic activity was observed with an IC 50 value of 81 μg/mL (22).…”