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. 2016 Oct 13;6(4):35.
doi: 10.3390/metabo6040035.

Metabolomic Profiling of the Effects of Melittin on Cisplatin Resistant and Cisplatin Sensitive Ovarian Cancer Cells Using Mass Spectrometry and Biolog Microarray Technology

Sanad Alonezi  1 , Jonans Tusiimire  2 , Jennifer Wallace  3 , Mark J Dufton  4 , John A Parkinson  5 , Louise C Young  6 , Carol J Clements  7 , Jin Kyu Park  8 , Jong Woon Jeon  9 , Valerie A Ferro  10 , David G Watson  11
Affiliations

Metabolomic Profiling of the Effects of Melittin on Cisplatin Resistant and Cisplatin Sensitive Ovarian Cancer Cells Using Mass Spectrometry and Biolog Microarray Technology

Sanad Alonezi et al. Metabolites. .

Abstract

In the present study, liquid chromatography-mass spectrometry (LC-MS) was employed to characterise the metabolic profiles of two human ovarian cancer cell lines A2780 (cisplatin-sensitive) and A2780CR (cisplatin-resistant) in response to their exposure to melittin, a cytotoxic peptide from bee venom. In addition, the metabolomics data were supported by application of Biolog microarray technology to examine the utilisation of carbon sources by the two cell lines. Data extraction with MZmine 2.14 and database searching were applied to provide metabolite lists. Principal component analysis (PCA) gave clear separation between the cisplatin-sensitive and resistant strains and their respective controls. The cisplatin-resistant cells were slightly more sensitive to melittin than the sensitive cells with IC50 values of 4.5 and 6.8 μg/mL respectively, although the latter cell line exhibited the greatest metabolic perturbation upon treatment. The changes induced by melittin in the cisplatin-sensitive cells led mostly to reduced levels of amino acids in the proline/glutamine/arginine pathway, as well as to decreased levels of carnitines, polyamines, adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (NAD+). The effects on energy metabolism were supported by the data from the Biolog assays. The lipid compositions of the two cell lines were quite different with the A2780 cells having higher levels of several ether lipids than the A2780CR cells. Melittin also had some effect on the lipid composition of the cells. Overall, this study suggests that melittin might have some potential as an adjuvant therapy in cancer treatment.

Keywords: A2780 cells; LC-MS; Melittin; cisplatin resistance; metabolomics; ovarian cancer.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effect of melittin on the viability of the ovarian cancer cells A2780 and A2780CR. Cell viability was determined following treatment with varying doses of melittin for 24 h (IC50 = 6.8 µg/mL A2780; IC50 = 4.5 µg/mL A2780CR).
Figure 2
Figure 2
Comparison of substrate metabolism in A2780 and A2780CR. Dye reduction rates calculated following 24 h incubation of cells.
Figure 3
Figure 3
Comparison of substrate metabolism in A2780 cells following melittin exposure. Dye reduction rates calculated following 24 h incubation of cells with melittin at IC50 (6.8 µg/mL) concentration. C = untreated controls; T = melittin treated.
Figure 4
Figure 4
Comparison of substrate metabolism in A2780CR cells following melittin exposure. Dye reduction rates calculated following 24 h incubation of cells with melittin at IC50 (4.5 µg/mL) concentration. C = untreated controls; T = melittin treated.
Figure 5
Figure 5
Multivariate data analysis of the ovarian cancer cells A2780 and A2780CR treated with melittin. PCA scores plot generated from PCA using LC-MS normalised data of treated cells and controls. MS circles: A2780-treated cells; C circles: untreated A2780 cells; MR circles: A2780CR -treated cells; CR circles: untreated A2780CR.
Figure 6
Figure 6
Heat Map showing the relative abundance of lipids in A2780 (S), A2780CR (R) and melittin treated (MS & MR) cells. Red = 5 × 107, Yellow = 8% of highest value, Blue = 0%.
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