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. 2016 Mar 31:7:89.
doi: 10.3389/fphar.2016.00089. eCollection 2016.

Dual Regulation of Cell Death and Cell Survival upon Induction of Cellular Stress by Isopimara-7,15-Dien-19-Oic Acid in Cervical Cancer, HeLa Cells In vitro

Nadiah Abu  1 , Swee K Yeap  2 , Ahmad Z Mat Pauzi  1 , M Nadeem Akhtar  3 , Nur R Zamberi  1 , Jamil Ismail  3 , Seema Zareen  3 , Noorjahan B Alitheen  1
Affiliations

Dual Regulation of Cell Death and Cell Survival upon Induction of Cellular Stress by Isopimara-7,15-Dien-19-Oic Acid in Cervical Cancer, HeLa Cells In vitro

Nadiah Abu et al. Front Pharmacol. .

Abstract

The Fritillaria imperialis is an ornamental flower that can be found in various parts of the world including Iraq, Afghanistan, Pakistan, and the Himalayas. The use of this plant as traditional remedy is widely known. This study aims to unveil the anti-cancer potentials of Isopimara-7,15-Dien-19-Oic Acid, extracted from the bulbs of F. imperialis in cervical cancer cell line, HeLa cells. Flow cytometry analysis of cell death, gene expression analysis via cDNA microarray and protein array were performed. Based on the results, Isopimara-7,15-Dien-19-Oic acid simultaneously induced cell death and promoted cell survival. The execution of apoptosis was apparent based on the flow cytometry results and regulation of both pro and anti-apoptotic genes. Additionally, the regulation of anti-oxidant genes were up-regulated especially thioredoxin, glutathione and superoxide dismutase- related genes. Moreover, the treatment also induced the activation of pro-survival heat shock proteins. Collectively, Isopimara-7,15-Dien-19-Oic Acid managed to induce cellular stress in HeLa cells and activate several anti- and pro survival pathways.

Keywords: 15-dien-19-oic acid; Fritillaria imperialis; HeLa; antioxidant; isopimara-7.

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Figures

Figure 1
Figure 1
The chemical structure of DIA isolated from the bulbs of Fritillaria imperialis.
Figure 2
Figure 2
MTT analysis of HeLa cells after being treated with DIA for 48 h at 30 μg/mL followed by seven 2-serial dilutions.
Figure 3
Figure 3
Flow cytometry analysis of (A) Cell cycle analysis by staining the DNA using propidium iodide, (B) Annexin V analysis for the detection of externalization of phosphatidylserine (LL, viable; LR, Early apoptosis; UR, Late apoptosis), and (C) JC-1 analysis for the detecting the change of mitochondrial membrane potential in HeLa cells (Red: Aggregates; Green: Monomers) after 48 h of treatment with 15 μg/mL of DIA.
Figure 4
Figure 4
Representative image of the proteome profiler analysis for cell stress-related images as well as the quantification values for HeLa cells; both control (untreated) and DIA-treated cells (15 ug/mL) for 48 h.
Figure 5
Figure 5
Validation of the proteome profiler results by comparing the expression of proteins to the respective genes in microarray (p-value for the microarray results are p < 0.05).
Figure 6
Figure 6
Validation of the microarray results using quantitative real-time PCR with selected genes in total RNA samples extracted from the control cells and 15 μg/mL DIA-treated cells (p-value for the microarray results are p < 0.05).
Figure 7
Figure 7
Bar chart analysis of the SOD units/mg of protein and μmol GSH/mg of protein in control cells and DIA-treated HeLa cells after 48 h of treatment with 15 μg/mL of DIA.
Figure 8
Figure 8
Proposed schematic of the mechanism of action of DIA in HeLa cells by regulating both the anti- and pro-survival pathways.
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