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. 2022 Sep 19;27(18):6106.
doi: 10.3390/molecules27186106.

Chemical and Antimicrobial Characterization of Mentha piperita L. and Rosmarinus officinalis L. Essential Oils and In Vitro Potential Cytotoxic Effect in Human Colorectal Carcinoma Cells

Alina Dolghi  1   2 , Dorina Coricovac  1   2 , Stefania Dinu  3 , Iulia Pinzaru  1   2 , Cristina Adriana Dehelean  1   2 , Cristina Grosu  1 , Doina Chioran  3 , Petru Eugen Merghes  4 , Cristian Andrei Sarau  5
Affiliations

Chemical and Antimicrobial Characterization of Mentha piperita L. and Rosmarinus officinalis L. Essential Oils and In Vitro Potential Cytotoxic Effect in Human Colorectal Carcinoma Cells

Alina Dolghi et al. Molecules. .

Abstract

Colorectal cancer is one of the most frequently diagnosed forms of cancer, and the therapeutic solutions are frequently aggressive requiring improvements. Essential oils (EOs) are secondary metabolites of aromatic plants with important pharmacological properties that proved to be beneficial in multiple pathologies including cancer. Mentha piperita L. (M_EO) and Rosmarinus officinalis L. (R_EO) essential oils are well-known for their biological effects (antimicrobial, antioxidant, anti-inflammatory and cytotoxic in different cancer cells), but their potential as complementary treatment in colorectal cancer is underexplored. The aim of the present study was to investigate the M_EO and R_EO in terms of chemical composition, antioxidant, antimicrobial, and cytotoxic effects in a colorectal cancer cell line-HCT 116. The gas-chromatographic analysis revealed menthone and menthol, and eucalyptol, α-pinene and L-camphor as major compounds in M_EO and R_EO respectively. M_EO exhibited potent antimicrobial activity, moderate antioxidant activity and a low cytotoxic effect in HCT 116 cells. R_EO presented a significant cytotoxicity in colorectal cancer cells and a low antimicrobial effect. The cytotoxic effect on non-cancerous cell line HaCaT was not significant for both essential oils. These results may provide an experimental basis for further research concerning the potential use of M_EO and R_EO for anticancer treatment.

Keywords: Mentha piperita L. essential oil; Rosmarinus officinalis L. essential oil; antimicrobial potential; colorectal cancer; cytotoxicity.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Viability assessment of Mentha piperita L. essential oil—M_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL) in HCT 116 cells at 24 h post-stimulation by MTT assay. The results are presented as cell viability percentage (%) normalized to control (non-stimulated) cells. These data represent the mean values ± SD of three independent experiments performed in triplicate. One-way ANOVA test was performed to determine the statistical differences in rapport with control group followed by Dunnett’s multiple comparisons post-test (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001).
Figure 2
Figure 2
Morphological aspect of HCT 116 cells after treatment for 24 h with Mentha piperita L. essential oil—M_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL).
Figure 3
Figure 3
Colorectal cancer cells—HCT 116 nuclei stained with Hoechst 33342 dye after a 24 h treatment with Mentha piperita L. essential oil—M_EO (10 and 150 μg/mL). Staurosporine (5 µM) was used as the positive control for apoptotic changes at nuclear level and Triton X-100 (0.5%) for necrosis. The yellow arrows indicate signs of apoptosis, and the red arrow shows signs of necrosis. The scale bar was 20 µm.
Figure 4
Figure 4
Viability assessment of Mentha piperita L. essential oil—M_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL) in HaCaT cells at 24 h post-stimulation by MTT assay. The results are presented as cell viability percentage (%) normalized to control (non-stimulated) cells. These data represent the mean values ± SD of three independent experiments performed in triplicate. One-way ANOVA test was performed to determine the statistical differences in rapport with control group followed by Dunnett’s multiple comparisons post-test (** p < 0.01, *** p < 0.001 and **** p < 0.0001).
Figure 5
Figure 5
Morphological aspect of HaCaT cells after treatment for 24 h with Mentha piperita L. essential oil—M_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL). The scale bar was 200 μm.
Figure 6
Figure 6
Viability assessment of Rosmarinus officinalis L. essential oil—R_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL) in HCT 116 cells at 24 h post-stimulation by MTT assay. The results are presented as cell viability percentage (%) normalized to control (non-stimulated) cells. These data represent the mean values ± SD of three independent experiments performed in triplicate. One-way ANOVA test was performed to determine the statistical differences in rapport with control group followed by Dunnett’s multiple comparisons post-test (*** p < 0.001 and **** p < 0.0001).
Figure 7
Figure 7
Morphological aspect of HCT 116 cells after treatment for 24 h with Rosmarinus officinalis L. essential oil—R_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL). The scale bar was 200 μm.
Figure 8
Figure 8
Colorectal cancer cells—HCT 116 nuclei stained with Hoechst 33342 dye after a 24 h treatment with Rosmarinus officinalis L. essential oil—R_EO (10 and 150 μg/mL). Staurosporine (5 µM) was used as the positive control for apoptotic changes at nuclear level and Triton X-100 (0.5%) for necrosis. The yellow arrows indicate signs of apoptosis, and the red arrow shows signs of necrosis. The scale bar was 20 µm.
Figure 9
Figure 9
Viability assessment of Rosmarinus officinalis L. essential oil—R_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL) in HaCaT cells at 24 h post-stimulation by MTT assay. The results are presented as cell viability percentage (%) normalized to control (non-stimulated) cells. These data represent the mean values ± SD of three independent experiments performed in triplicate. One-way ANOVA test was performed to determine the statistical differences in rapport with control group followed by Dunnett’s multiple comparisons post-test (* p < 0.05, ** p < 0.01 and **** p < 0.0001).
Figure 10
Figure 10
Morphological aspect of HaCaT cells after treatment for 24 h with Rosmarinus officinalis L. essential oil—R_EO (10, 50, 100, 150, 200, 250 and 500 μg/mL). The scale bar was 200 μm.
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