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, 17 (1), 252

Melicope Ptelefolia Leaf Extracts Exhibit Antioxidant Activity and Exert Anti-Proliferative Effect With Apoptosis Induction on Four Different Cancer Cell Lines

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Melicope Ptelefolia Leaf Extracts Exhibit Antioxidant Activity and Exert Anti-Proliferative Effect With Apoptosis Induction on Four Different Cancer Cell Lines

Mohammad Faujul Kabir et al. BMC Complement Altern Med.

Abstract

Background: Melicope ptelefolia is a well-known herb in a number of Asian countries. It is often used as vegetable salad and traditional medicine to address various ailments. However, not many studies have been currently done to evaluate the medicinal benefits of M. ptelefolia (MP). The present study reports antioxidant, anti-proliferative, and apoptosis induction activities of MP leaf extracts.

Method: Young MP leaves were dried, powdered and extracted sequentially using hexane (HX), ethyl acetate (EA), methanol (MeOH) and water (W). Antioxidant activity was evaluated using ferric reducing antioxidant power (FRAP), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) radicals scavenging and cellular antioxidant activity (CAA) assays. Anti-proliferative activity was evaluated through cell viability assay, using the following four human cancer cell lines: breast (HCC1937, MDA-MB-231), colorectal (HCT116) and liver (HepG2). The anti-proliferative activity was further confirmed through cell cycle and apoptosis assays, including annexin-V/7-aminoactinomycin D staining and measurements of caspase enzymes activation and inhibition.

Result: Overall, MP-HX extract exhibited the highest antioxidant potential, with IC50 values of 267.73 ± 5.58 and 327.40 ± 3.80 μg/mL for ABTS and DPPH radical-scavenging assays, respectively. MP-HX demonstrated the highest CAA activity in Hs27 cells, with EC50 of 11.30 ± 0.68 μg/mL, while MP-EA showed EC50 value of 37.32 ± 0.68 μg/mL. MP-HX and MP-EA showed promising anti-proliferative activity towards the four cancer cell lines, with IC50 values that were mostly below 100 μg/mL. MP-HX showed the most notable anti-proliferative activity against MDA-MB-231 (IC50 = 57.81 ± 3.49 μg/mL) and HCT116 (IC50 = 58.04 ± 0.96 μg/mL) while MP-EA showed strongest anti-proliferative activity in HCT116 (IC50 = 64.69 ± 0.72 μg/mL). The anticancer potential of MP-HX and MP-EA were also demonstrated by their ability to induce caspase-dependent apoptotic cell death in all of the cancer cell lines tested. Cell cycle analysis suggested that both the MP-HX and MP-EA extracts were able to disrupt the cell cycle in most of the cancer cell lines.

Conclusions: MP-HX and MP-EA extracts demonstrated notable antioxidant, anti-proliferative, apoptosis induction and cancer cell cycle inhibition activities. These findings reflect the promising potentials of MP to be a source of novel phytochemical(s) with health promoting benefits that are also valuable for nutraceutical industry and cancer therapy.

Keywords: Anticancer; Antioxidant activity; Apoptosis; Cell cycle; Cellular antioxidant assay; HCC1937; HCT116; HepG2; MDA-MB-231; Melicope ptelefolia.

Figures

Fig. 1
Fig. 1
Determination of antioxidant components and in vitro antioxidant activities of MP leaf extracts. (a) Total phenolic content, (b) Total flavonoid content, (c) ABTS radical-scavenging, and (d) DPPH radical-scavenging. Values are mean ± SD (n = 3). RB, raspberry; BKB, blackberry; BLB, blueberry; HX, hexane; EA, ethyl acetate; MeOH, methanol; W, water. Means with different letters (a-d) are significantly different (p < 0.05). GAE, gallic acid equivalent; QE, quercetin equivalent; DE, dried extract; Tx, trolox; AA, ascorbic acid; Cn, catechin; Qn, quercetin
Fig. 2
Fig. 2
Cellular antioxidant activity of MP leaf extracts. Inhibition of peroxyl radical-induced oxidation of DCFH2 in Hs27 cells by a MP-HX, b MP-EA, c MP-MeOH, d MP-W, e Quercetin f Polyphenon-60 and g Trolox. The curves shown in each graph are from a single experiment (mean ± SD, n = 3)
Fig. 3
Fig. 3
Dose-response curve for inhibition of peroxyl radical-induced DCFH2 oxidation in Hs27 cells by MP leaf extracts and positive controls. a MP-HX, b MP-EA, c MP-MeOH, d MP-W, e Quercetin, f Polyphenon-60 and g Trolox. The curves shown in each graph are from a single experiment (mean ± SD, n = 3)
Fig. 4
Fig. 4
MTS cell viability assay. Dose response curves for the cytotoxic effect of MP-HX and MP-EA (48 h treatment) on cancerous (HCT116, HCC1937, HepG2, MDA-MB-231) and non-cancerous (CCD841, Hs27) cell lines
Fig. 5
Fig. 5
The effect of MP-HX or MP-EA on caspase 3/7 activity in cancer cell lines. The bar charts depict luminescence value for caspase 3/7 activity. The concentration of the extracts used in each treatment was at ~IC50 value for the corresponding cell lines indicated. T24/T48, 24 or 48 h treatment, respectively. VC, vehicle control; FC, fold change compared to VC. Values are mean ± SD (n = 3). The values with different letters (a-d) are significantly different, p < 0.05
Fig. 6
Fig. 6
Cell viability and multicaspase enzyme activation assays. (a) The plots depict the effect of MP-HX and MP-EA treatments in the cancer cell lines indicated. Each plot is a representative figure of the three replicates of each determination. (b) The bar charts depict the percentage of live cells and those with multicaspase enzyme activation. The concentration of the extracts used in each treatment was at ~IC50 value for the corresponding cell lines indicated. T24/T48, 24 or 48 h treatment, respectively. VC, vehicle control. Values are mean ± SD (n = 3). The values with different letters (a-d) are significantly different, p < 0.05
Fig. 7
Fig. 7
Annexin-V & Dead Cell (7-AAD) flow cytometry analysis. Apoptotic effect of MP-HX and MP-EA on HCT116, HCC1937, HepG2 and MDA-MB-231 cell lines. (a) Apoptosis profile plot. Each plot is a representative figure of the three replicates of each determination. (b) Bar charts depicting percentage of live, dead and apoptotic cells for the treatments on the corresponding cell lines. Values are mean ± SD (n = 3). The values with different letters (a-d) are significantly different, p < 0.05. The concentration of the extracts used in each treatment was at ~IC50 value for the corresponding cell lines indicated. T24, 24 h treatment; T48, 48 h treatment; VC, vehicle control. L, live cells; EA, early apoptotic cells, LA, late apoptotic cells; D, dead cells
Fig. 8
Fig. 8
Pan caspase inhibitor assay. MTS cell viability assay (48 h) was employed using MP-HX or MP-EA extracts on the cell lines indicated, in the presence or absence of Z-VAD-FMK inhibitor. The concentration of the extracts used in each treatment was at ~IC50 value for the corresponding cell lines indicated. Values are mean ± SD (n = 3). The values with different letters (a-c) are significantly different, p < 0.05
Fig. 9
Fig. 9
Effect of MP-EA and MP-HX on HCT116, HCC1937, HepG2 and MDA-MB-231 cell cycle distribution. The cells were treated for 24 h and the concentration of the extracts in each treatment was at ~IC50 value of the corresponding cell lines indicated. (a) Plot for DNA content profile. Each plot is a representative figure of the three replicates of each determination. (b) Bar charts depict the percentage of cells in the various stages of the cell cycle. Values are mean ± SD (n = 3). The values with different letters (a-c) are significantly different, p < 0.05

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References

    1. Rates SMK. Plants as a source of drugs. Toxicon. 2001;39:603–613. doi: 10.1016/S0041-0101(00)00154-9. - DOI - PubMed
    1. Halliwell B. Free radicals and antioxidants: updating a personal view. Nutr Rev. 2012;70(5):257–265. doi: 10.1111/j.1753-4887.2012.00476.x. - DOI - PubMed
    1. Rukayah A. Tumbuhan liar berkhasiat ubatan, Cet. 1. Kuala Lumpur: Dewan Bahasa dan Pustaka; 2006.
    1. Perry LM, Metzger J. Medicinal plants of East and Southeast Asia: attributed properties and uses. Cambridge: MIT Press; 1980.
    1. Sulaiman MR, Mohd Padzil A, Shaari K, Khalid S, Shaik Mossadeq WM, Mohamad AS, Ahmad S, Akira A, Israf D, Lajis N. Antinociceptive activity of Melicope ptelefolia ethanolic extract in experimental animals. J Biomed Biotechnol. 2010;2010:937642. doi: 10.1155/2010/937642. - DOI - PMC - PubMed
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