Toxicological aspect of bioinsecticide pyrethrum extract and expressions of apoptotic gene levels in human hepotacellular carcinoma HepG2 cells

Abstract

Pyrethrum extract (PE), an important natural bioinsecticide, is extensively used across the world to control pest insects in homes and farms. The aim of this study was to evaluate the potential cytotoxic effect of PE using MTT assay and genotoxic effect using micronucleus (MN) assay. The changes in the expressions of the apoptosis genes in mRNA levels were also investigated using Real-Time qPCR analysis as well as the ratio of apoptotic/necrotic cells with AnnexinV-FITC/Propidium iodide (PI) assay in HepG2 cells. PE markedly suppressed the cell proliferation on HepG2 cells. It significantly increased the frequency of micronucleus (MN) at 500 and 1000 µg/mL. PE also induced the percentage of the cell population of late apoptotic/necrotic cells (FITC + PI+) and necrotic cells (FITC- PI+), especially at 4000 μg/mL analyzed by flow cytometry. PE caused significant fold changes in the expression of several apoptotic genes including APAF1, BIK, BAX, BAD, BİD, MCL-1, CASP3, CASP1, CASP2, FAS, FADD and TNFRSF1A. In particular, the pro-apoptotic gene Hrk (Harakiri) remarkably and dose-dependently was overexpressed of the mRNA level. As a result, PE may exhibit cyto-genotoxic effects, especially at higher concentrations and lead to significant changes in the expression of mRNA levels in several apoptotic genes.

Highlights

• Natural bioinsecticide PE exhibited a cytotoxic effect in HepG2 cells.

• PE significantly induced the micronucleus (MN) frequency at 500 and 1000 µg/mL.

• This bioinsecticide induced cell death and it lead to significant fold changes in the expression of mRNA levels in several apoptotic genes in HepG2 cells.

• The highest increase of the expression of mRNA levels was determined in Hrk (Harakiri) at 4000 µg/mL.

Keywords:

• Pyrethrum extract
• MTT
• micronucleus
• Annexin-V/FITC-PI
• real time-qPCR
• HepG2 cells

Acknowledgement

This work was supported by Gazi University Research Fund (The Scientific Research Projects Coordination Unit) under Project number: 64/2018-06, Turkey. We would like to thank Prof. Dr. Ümit Emin Bağrıaçık for laboratory support and Dr. Handan Kayhan for flow cytometry analysis.

Disclosure statement

The authors declare no conflicts of interest.

Additional information

Funding

This work was supported by Gazi University Research Fund (The Scientific Research Projects Coordination Unit) under Project number: 64/2018-06, Turkey.