Exploring the Anticancer Potential of Origanum majorana Essential Oil Monoterpenes Alone and in Combination against Non-Small Cell Lung Cancer
<p>Chemical structure of the most abundant oxygenated monoterpenes and monoterpene hydrocarbons in OMEO (National Centre for Biotechnology Information. PubChem Compound Summary).</p> "> Figure 2
<p>Effect of the four monoterpenes on NSCLC cellular viability in vitro. A549 (<b>A</b>–<b>D</b>) and LNM35 (<b>E</b>–<b>H</b>) cells were treated with increasing concentrations of terpinen-4-ol, sabinene hydrate, α-terpinene, or <span class="html-italic">γ</span>-terpinene for 24, 48, and 72 h. Cellular viability was determined as described in the materials and methods. Experiments were repeated at least three times. Shapes represent mean and bars represent S.E.M. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, and **** <span class="html-italic">p</span> < 0.0001; ns—non-significant.</p> "> Figure 3
<p>Effect of the four monoterpenes on NSCLC colony growth. A549 and LNM35 formed colonies were treated every three days for 1 week with terpinen-4-ol (<b>A</b>,<b>E</b>), sabinene hydrate (<b>B</b>,<b>F</b>), α-terpinene (<b>C</b>,<b>G</b>), or <span class="html-italic">γ</span>-terpinene (<b>D</b>,<b>H</b>) for 1 week. Colonies were fixed, stained, and counted as described in the materials and methods. Experiments were repeated at least three times. Columns represent mean; bars represent S.E.M. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, and **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 4
<p>Kinetic effect of OMEO on survivin expression in NSCLC cells in vitro. Western blot analysis shows the level of survivin in A549 and LNM35 cells treated with OMEO 100 µg/mL for 0.5, 2, 6, 24, and 48 h. Experiments were repeated for at least three times.</p> "> Figure 5
<p>Effect of the four monoterpenes on survivin expression in NSCLC cells in vitro. A549 and LNM35 cells were treated with (<b>A</b>) terpinen-4-ol 0.05%, (<b>B</b>) sabinene hydrate 0.05%, (<b>C</b>) α-terpinene 0.02%, or (<b>D</b>) <span class="html-italic">γ</span>-terpinene 0.1% for 24 and 48 h. Experiments were repeated at least three times.</p> "> Figure 6
<p>Effect of monoterpene combinations on NSCLC cellular viability in vitro. A549 cells were treated with different concentrations of monoterpenes alone or in combination for 48 h. Cellular viability was assessed using CellTiter-Glo Luminescent Assay as described in the materials and methods. Experiment was repeated at least three times. Columns represent means, and bars represent S.E.M. **** <span class="html-italic">p</span> < 0.0001. ns—non-significant. Black stars indicate significance in comparison to the control; red stars indicate significance between groups.</p> "> Figure 7
<p>Effect of terpinen-4-ol in combination with sabinene hydrate on A549 viability and colony growth in vitro. (<b>A</b>–<b>C</b>) A549 cells were treated with terpinen-4-ol, sabinene hydrate, or a combination for 6, 24, and 48 h. Cellular viability was determined as described in the materials and methods. (<b>D</b>–<b>F</b>) A549 number of total and large colonies treated with terpinen-4-ol, sabinene hydrate, or a combination for 7 days. Experiments were repeated at least three times. Columns represent mean and bars are S.E.M. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, and **** <span class="html-italic">p</span> < 0.0001. ns—non-significant.</p> "> Figure 8
<p>Effect of terpinen-4-ol in combination with sabinene hydrate on LNM35 viability and colony growth in vitro. (<b>A</b>) LNM35 cells were treated with terpinen-4-ol, sabinene hydrate, or a combination for 6 h. Cellular viability was determined as described in the materials and methods. (<b>B</b>–<b>D</b>) LNM35 number of total and large colonies treated with terpinen-4-ol, sabinene hydrate, or a combination for 7 days. Experiments were repeated at least three times. Columns represent mean and bars are S.E.M. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, and **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 9
<p>Effect of the combination of terpinen-4-ol and sabinene hydrate on survivin expression. Quantification of the Western blot showing the impact of terpinen-4-ol + sabinene hydrate on the expression of survivin in A549 and LNM35 cells. Columns represent means of at least three independent experiments; bars represent S.E.M. * <span class="html-italic">p</span> < 0.05. ** <span class="html-italic">p</span> < 0.01. *** <span class="html-italic">p</span> < 0.001. ns—non-significant.</p> "> Figure 10
<p>Effect of the combination of terpinen-4-ol and sabinene hydrate on NSCLC tumor growth in CAM. (<b>A</b>,<b>B</b>) Weight of A549 tumors xenografted on CAM and treated with or without terpinen-4-ol, sabinene hydrate, or a combination for 1 week. (<b>C</b>) Percentage of alive embryos in the A549 control and treated groups. (<b>D</b>,<b>E</b>) Weight of LNM35 tumors xenografted on CAM and treated with or without terpinen-4-ol, sabinene hydrate, or a combination for 1 week. (<b>F</b>) Percentage of alive embryos in the LNM35 control and treated groups. Columns are means; bars are S.E.M. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, and **** <span class="html-italic">p</span> < 0.0001. ns—non-significant.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Reagents
2.2. Cellular Viability
2.3. Colony Growth Assay
2.4. In Ovo Tumor Growth Assay
2.5. Western Blot
2.6. Statistical Analysis
3. Results and Discussions
3.1. The Four Monoterpenes Inhibited NSCLC Cellular Viability and Colony Growth In Vitro
3.2. The Four Monoterpenes Inhibited Survivin Expression In Vitro
3.3. The Anticancer Effect of Monoterpene Combinations on NSCLC In Vitro
3.3.1. Impact of Terpinen-4-ol in Combination with Sabinene Hydrate on NSCLC Cellular Viability and Colony Growth In Vitro
3.3.2. Terpinen-4-ol in Combination with Sabinene Hydrate Decreased Survivin Expression
3.3.3. Impact of Terpinen-4-ol in Combination with Sabinene Hydrate on NSCLC Tumors In Vivo
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Arafat, K.; Al-Azawi, A.M.; Sulaiman, S.; Attoub, S. Exploring the Anticancer Potential of Origanum majorana Essential Oil Monoterpenes Alone and in Combination against Non-Small Cell Lung Cancer. Nutrients 2023, 15, 5010. https://doi.org/10.3390/nu15235010
Arafat K, Al-Azawi AM, Sulaiman S, Attoub S. Exploring the Anticancer Potential of Origanum majorana Essential Oil Monoterpenes Alone and in Combination against Non-Small Cell Lung Cancer. Nutrients. 2023; 15(23):5010. https://doi.org/10.3390/nu15235010
Chicago/Turabian StyleArafat, Kholoud, Aya Mudhafar Al-Azawi, Shahrazad Sulaiman, and Samir Attoub. 2023. "Exploring the Anticancer Potential of Origanum majorana Essential Oil Monoterpenes Alone and in Combination against Non-Small Cell Lung Cancer" Nutrients 15, no. 23: 5010. https://doi.org/10.3390/nu15235010