Natural Compounds Modulate Drug Transporter Mediated Oral Cancer Treatment
<p>ATP-binding cassette G2 (ABCG2) expression was different in varied head and neck cancer (HNC) cell lines, which affected the amount of protoporphyrin IX (PpIX) and the survival percentage while the cells were treated with 5-aminolevulinic acid photodynamic therapy (ALA-PDT), and was modulated by different compounds. (<b>A</b>) Western blot analysis showed that ABCG2 was highly expressed in OECM1 but not in SAS, HSC3, and Fadu cells. (<b>B</b>) In parental status, PpIX amount of the four HNC cell lines showed no significant difference. For those cells treated with ALA, OECM1 produced the lowest amount of PpIX in comparison with SAS, HSC3, and Fadu cells. Switch of SAS cells from a medium containing 25 mM glucose to 5.5 mM glucose (<b>C</b>) induced ABCG2 expression and (<b>D</b>) downregulated PpIX accumulation in a time-dependent manner. (<b>E</b>) Western blot analysis for phospho-Akt, total Akt, Nrf2, and ABCG2 protein expression in OECM1 and SASL90d cells with/without treatment of PI3K inhibitor LY294002 (<span class="html-italic">n</span> ≥ 3; *<span class="html-italic">p</span> < 0.05; ***<span class="html-italic">p</span> < 0.001). IC<sub>50</sub> of ALA-PDT treatment was measured in ABCG2-enriched cells: (<b>F</b>) OECM1, parental SAS, and SASL90d cells; (<b>G</b>) OECM1; and (<b>H</b>) SASL90d. ABCG2 level was significantly inhibited by gefitinib, epigallocatechin gallate (EGCG), and curcumin but not by naringenin. <span class="html-italic">n</span> ≥ 3, *<span class="html-italic">p</span> < 0.05, ***<span class="html-italic">p</span> < 0.001, ****<span class="html-italic">p</span> < 0.0001.</p> "> Figure 2
<p>Gefitinib treatment suppresses ABCG2 expression, increases PpIx accumulation, and decreases ALA-PDT-mediated HNC cell viability. (<b>A</b>) Western blot analysis for p-EGFR (Y1068), p-Akt (S473), Nrf2, and ABCG2 expression in OECM1 and SASL90d cells treated with different concentrations of gefitinib. Gefitinib repressed EGFR and Akt signaling activities as well as ABCG2 and Nrf2 expression. In OECM1 and SASL90d, gefitinib treatment (<b>B</b>) upregulated PpIX accumulation and (<b>C</b>) decreased cell viability in a dose-dependent manner. <span class="html-italic">n</span> ≥ 3; *<span class="html-italic">p</span> < 0.05; **<span class="html-italic">p</span> < 0.01; ***<span class="html-italic">p</span> < 0.001.</p> "> Figure 3
<p>EGCG treatment suppresses ABCG2 expression, increases PpIx accumulation, and decreases ALA-PDT mediated-HNC cell viability. (<b>A</b>) Western blot analysis for p-EGFR (Y1068), p-Akt (S473), Nrf2, and ABCG2 expression in OECM1 and SASL90d cells treated with different concentrations of tea polyphenol EGCG. EGCG inhibited EGFR and Akt signaling activities as well as ABCG2 and Nrf2 expression. In OECM1 and SASL90d, EGCG treatment (<b>B</b>) upregulated PpIX accumulation and (<b>C</b>) decreased cell viability in a dose-dependent manner. <span class="html-italic">n</span> ≥ 3; ***<span class="html-italic">p</span> < 0.001, ****<span class="html-italic">p</span> < 0.0001.</p> "> Figure 4
<p>Curcumin treatment suppresses ABCG2 expression, increases PpIx accumulation, and decreases ALA-PDT-mediated HNC cell viability. (<b>A</b>) Western blot analysis for p-EGFR (Y1068), p-Akt (S473), Nrf2, and ABCG2 expression in OECM1 and SASL90d cells treated with different concentrations of curcumin. Curcumin inhibited EGFR and Akt signaling activities as well as ABCG2 and Nrf2 expression. In OECM1 and SASL90d, curcumin treatment (<b>B</b>) upregulated PpIX accumulation and (<b>C</b>) decreased cell viability in a dose-dependent manner. <span class="html-italic">n</span> ≥ 3; *<span class="html-italic">p</span> < 0.05; **<span class="html-italic">p</span> < 0.01, ****<span class="html-italic">p</span> < 0.0001.</p> "> Figure 5
<p>ABCG2 expression is associated with increased stemness, tumor grade, and HNC prognosis. (<b>A</b>) ABCG2 was enriched in SAS sphere culture compared with SAS parental cells. (<b>B</b>) Aldehyde dehydrogenase (ALDH)<sup>+</sup> cell population gradually increased once SAS cells were cultured in a medium containing lower glucose. ABCG2 expression increased with increasing low-glucose culture time. ALDH<sup>+</sup> percentage also increased with ABCG2 expression, with individual values of 16.24%, 18.29%, and 27.01%. BODIPY-aminoacetaldehyde (BAAA) is an ALDH substrate, and diethylaminobenzaldehyde (DEAB) is an inhibitor of ALDH activity. (<b>C</b>) Statistical analysis for ABCG2 mRNA levels in normal and primary HNC tissues stratified by clinical grades from UALCAN database. *<span class="html-italic">p</span> < 0.05. (<b>D</b>) Kaplan–Meier analysis for cancer-specific survival rates in HNSCC patients classified by ABCG2 expression using the Human Protein Atlas database. The expression cut-off value is 0.32 FPKM (fragments per kilobase of transcript per million mapped reads).</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Lines and Culture
2.2. ALA-PDT
2.3. 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Assay
2.4. Western Blot Analysis
2.5. PpIX Detection
2.6. Primary Tumor Sphere Culture
2.7. ABCG2 Detection
2.8. Aldehyde dehydrogenase (ALDH) Assay
2.9. Statistical Analysis
3. Results
3.1. PpIX Accumulation Inversely Correlates with ABCG2 Expression
3.2. Gefitinib, Curcumin, and EGCG Inhibited ABCG2 Expression and Modulated PpIX Accumulation and ALA-PDT Efficiency
3.3. ABCG2 Expression Correlated with Stemness, Tumor Grade, and Clinical Prognosis of HNCs
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Yang, H.; Wei, Y.-C.; Li, W.-C.; Chen, H.-Y.; Lin, H.-Y.; Chiang, C.-P.; Chen, H.-M. Natural Compounds Modulate Drug Transporter Mediated Oral Cancer Treatment. Biomolecules 2020, 10, 1335. https://doi.org/10.3390/biom10091335
Yang H, Wei Y-C, Li W-C, Chen H-Y, Lin H-Y, Chiang C-P, Chen H-M. Natural Compounds Modulate Drug Transporter Mediated Oral Cancer Treatment. Biomolecules. 2020; 10(9):1335. https://doi.org/10.3390/biom10091335
Chicago/Turabian StyleYang, Hsiang, Yu-Ching Wei, Wan-Chun Li, Hsin-Yung Chen, Hung-Ying Lin, Chun-Pin Chiang, and Hsin-Ming Chen. 2020. "Natural Compounds Modulate Drug Transporter Mediated Oral Cancer Treatment" Biomolecules 10, no. 9: 1335. https://doi.org/10.3390/biom10091335