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Combination of anticancer drugs with quercetin in human lung cancer cells

Combination of anticancer drugs with quercetin in human lung cancer cells. Chemotherapy is a common strategy used to kill cancer cells. However toxicity of chemotherapy drugs to normal cells limits their application.

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Combination of anticancer drugs with quercetin in human lung cancer cells

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  1. Combination of anticancer drugs with quercetin in human lung cancer cells

  2. Chemotherapy is a common strategy used to kill cancer cells. • However toxicity of chemotherapy drugs to normal cells limits their application.

  3. Several phytochemicals, such as quercetin, have been reported to prevent cancer development by themselves or by enhancing the effects of anticancer drugs. • For example, it has been shown that quercetin (10–40 mM) in combination with Trichostatin A (TSA) cooperatively induces cell death in human leukemia HL-60 cells (Chen and Kang, 2005)

  4. TSA and vorinostat are histone deacetylase inhibitors (HDI), which are a promising class of anticancer drugs because they selectively induce the differentiation and apoptosis of various transformed cells. TSA Vorinostat

  5. Quercetin, a flavonoid, is a phytochemical found in various vegetal foods, such as onions, apples, and green leafy vegetables. It has been suggested to possess anti-oxidative and antiinflammatory Properties. quercetin

  6. Besides the synergistic or additive inhibiting effects, the combined treatment also reduces the toxicity of chemotherapy due to the lower dose of each compound.

  7. Does quercetin affect the anticancer effect of TSA on human lung cancer cells?

  8. In vitro study A549 cells (p53+/+) or H1299 cells (p53-/-) TSA (25 ng/mL) aloneor combined with quercetin (2 or 5 μM) 1. Cell growth 2. Apoptosis 3. p53, Bax, Apaf-1, Bcl-2, cytochrome c, acetyl-H3/H4 protein expression 4. caspase-9/3 activity

  9. Quercetin has a stronger enhancing effect on TSA-induced cell-growth arrest and apoptosis in A549. H1299 A549 A549 H1299

  10. Quercetin increases p53 protein expression in A549 cells. (A) p53 GAPDH (B)

  11. Quercetin enhances TSA-induced apoptosis through the mitrochondria pathway in A549 cells. control TSA 5Q 5Q+TSA Bax Apaf-1 GAPDH control TSA 5Q 5Q+TSA Cytochrome C (cytosol) GAPDH (cytosol) Cytochrome C (mitochondria) GAPDH (mitochondria) caspase-3 caspase-9

  12. (A) + p53 siRNA nontargeted control control TSA 5Q 5Q+TSA Bax Apaf-1 GAPDH (B) caspase-9 caspase-3

  13. These results indicate that regulation of the expression of p53 protein plays an important role in enhancing TSA-induced apoptosis in A549 cells.

  14. In Vivo study lls

  15. (B) IHC Staining for p53 Quercetin administrated by IP injection enhances anticancer effect of TSA in tumor-bearing mice. (A) control LT HT LT+Q p53 GAPDH LT control HT LT+Q

  16. TSA Quercetin Quercetin enhances the anticancer effect of TSA in vitro and in vivo possibly through a p53-dependent pathway. H1299 apoptosis associated gene p300 apoptosis

  17. Does quercetin given orally also has the same effect ? Nude mice ？ Nude mice oral TSA Quercetin Nude mice Nude mice Nude mice Enhancing effect Nude mice i.p. injection

  18. It is known that after quercetin intake, conjugated metabolites, such as quercetin glucuronides and quercetin sulfates rather than quercetin aglycone are mainly present in human plasma. HPLC chromatograms in the plasma of nude mice.

  19. In vivo study Week 2：subcutaneously injected A549 cells : 5x106 cells Week 20 animals were sacrificed and analyzed acclimated for 1 week Week 5：Randomly assigned (6 groups) Control TSA : 0.5 mg/kg B.W., 2 times/week, i.p. OL : 20 mg/kg B.W., 3 times/week, gavage OH : 100 mg/kg B.W., 3 times/week, gavage IL : 2 mg/kg B.W., 3 times/week, i.p. IH : 10 mg/kg B.W., 3 times/week, i.p. Nude mice

  20. Quercetin given orally at the doses, 20 and 100 mg/kg 3 times/week, fail to enhance the anticancer effect of TSA.

  21. The contributing factor for the ineffectiveness of oral quercetin administration could be associated with the metabolic conversion of quercetin in vivo. Plasma Tumor tissues Total or individual concentrations of quercetin and its metabolites in plasma (A) and (B) tumor tissues of tumor-bearing mice.

  22. In vitro study Q3G significantly enhances the antiproliferation effect of TSA in A549 cells, however, the enhancing effect is less than that of quercetin.

  23. Quercetin given orally decreases the oxidative stress induced by TSA. The effect is similar to that of quercetin given by i.p. injection. C TSA (A) Lymphocyte DNA Damage (B) Lipid peroxidation IH+TSA OH+TSA

  24. Summary The study showed that quercetin given by gavage (20 and100 mg/kg body weight, 3 times/week;about 0.0015 and 0.0075 g/week) does not enhance the antitumor effect of TSA in tumor bearing mice. We postulate that the oral quercetin doses used are too low to exert the enhancing effect, because quercetin-3-glucuronide still has some enhancing effect on TSA-induced apoptosis.

  25. In vivo study Week 2：subcutaneously injected A549 cells : 5x106 cells Week 19 animals were sacrificed and analyzed acclimated for 1 week Week 4：Randomly assigned (5 groups) Control TSA : 0.5 mg/kg B.W., 2 times/week, i.p. OL : 0.1% quercetin diet OH : 1% quercetin diet IQ : 10 mg/kg B.W., 3 times/week, i.p. Nude mice These diet gave quercetin about 5-, 50-fold compared with the last study.

  26. Similar to IQ+TSA, OH+TSA significantly decreases tumor growth in A549-tumor-bearing nude mice. C TSA OL OH IQ p53 (A) (B) Tumor tissues GAPDH TSA+ TSA OH+TSA IQ+TSA

  27. (A) (B) OL and OH decrease TSA-induced DNA damage and muscle weight loss.

  28. Does quercetin enhance the anticancer effect of cisplatin, a widely used chemotherapy drug?

  29. In vivo study Week 2：subcutaneously injected A549 cells : 5x106 cells Week 20 animals were sacrificed and analyzed acclimated for 1 week Week 5：Randomly assigned (6 groups) Control cisplatin : 2 or 5 mg/kg, 1 time/week, i.p. OL : 0.1% quercetin diet OH : 1% quercetin diet IQ : 10 mg/kg B.W., 3 times/week, i.p. Nude mice

  30. Cis2 in combination with HQ and IQ rather than Cis2 alone significantly inhibit tumor growth. The combined effects are similar to that of cis5. CIS2 CIS2+HQ CIS2+IQ CIS5

  31. In A549 cells Both Quercetin and quercetin glucuronide significantly enhance the antiproliferation effect of cisplatin (1 μM). Quercetin Quercetin-3-glucuronide

  32. CIS markedly induces cell cycle arrest in G2/M phase. CIS CIS+Q increases the cells in sub-G1phase (apoptosis). CIS+Q Q3G only enhances the cell cycle arrest effect of CIS. CIS+Q3G

  33. C CIS 5Q 5G CIS +5Q CIS +5G 12 h P21 GAPDH 24 h P21 GAPDH P53 GAPDH P53 GAPDH 12 h 24 h Cisplatin in combination with quercetin or quercetin glucuronide increases the p21 and p53 protein expression. Consistent with our previous results, the efficiency of quercetin is better than that of quercetin glucuronide.

  34. Conclusion • Quercetin given orally or by i.p. injection may enhance the anticancer effect of TSA or cisplatin in vivo. • The enhancing effects of quercetin given orally are less than that by i.p. injection, because of the metabolic conversion. • The mechanisms by which quercetin exerts its effect are associated with upregulation of p21 and p53 expression in human lung cancer cells.

  35. Acknowledgement Dr. Shu-Ting Chan

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