Laporkan Masalah

The Anticancer Properties from Curcumin Analogs Pentagamavunone-1 (PGV-1) and Chemoprevention Curcumin Analog 1.1 (CCA-1.1) Against Aggressive Breast Cancer Cells

DHANIA NOVITASARI, Prof. Dr. apt. Edy Meiyanto, M.Si; Dr. apt. Riris Istighfari Jenie, M.Si; Prof. Jun-ya Kato, Ph.D.

2022 | Disertasi | DOKTOR ILMU FARMASI

Among breast cancer subtypes, triple-negative breast cancer (TNBC) and human epidermal growth factor receptor 2 (HER2)-positive breast cancer are aggressive types with poor prognosis. Curcumin analogs pentagamavunone-1 (PGV-1) and chemoprevention curcumin analog 1.1 (CCA-1.1) have been explored for anticancer properties against leukemia, breast cancer, and colorectal cancer cells. The present study aimed to evaluate the antitumor activities of PGV-1 and CCA-1.1 in altering the cellular physiology of TNBC and HER2-positive breast cancer cells. This study used MDA-MB-231 (TNBC) and HCC1954 (HER2-positive) cancer cells. The trypan blue exclusion assay was used to determine the cytotoxic effect of PGV-1 and CCA-1.1. Tumor growth inhibition was assessed using a xenograft mice model. The cell cycle profile was examined via flow cytometry, and the mitotic cells were quantified using Hoechst staining. Protein level expression of mitotic kinases (p-cyclin B1, p-PLK1, p-Aurora A, and Aurora A) was determined using western blotting. The intracellular reactive oxygen species (ROS) level was measured via flow cytometry, whereas mitochondrial respiration was monitored with a flux analyzer. The senescence phenomenon was observed using X-gal staining, and the antimigratory activity was monitored using wound healing assay. Then, the secretion level of matrix metalloproteinase 2 and 9 (MMP-2 and MMP9) was examined using gelatin zymography. PGV-1 and CCA-1.1 exhibited cytotoxic activity with GI 50 values of 0.9 and 1.4 µM in MDA-MB-231 cells, respectively. Moreover, they had a cytotoxic effect on HCC1954 cells with GI 50 values of 0.4 and 1.2 µM, respectively. PGV-1 and CCA-1.1 suppressed cell growth with irreversible effects. Oral administration of PGV-1 suppressed tumor growth in mice. The cell cycle profile indicated that PGV-1 induced prometaphase arrest and CCA-1.1 promoted metaphase arrest. Subsequently, PGV-1 tended to inhibit Aurora A kinase in MDA-MB-231 cells and induce cyclin B1 and PLK1 phosphorylation in MDA-MB-231 and HCC1954 cells. CCA-1.1 tended to inhibit Aurora A kinase in MDA-MB-231 cells but not in HCC1954 cells and increase PLK1 and cyclin B1 phosphorylation in HCC1954 cells. The ROS level increased after treatment with PGV-1 or CCA-1.1; this activity might impair mitochondrial function and induce senescence. PGV-1 inhibited MDA-MB-231 and HCC1954 cell migration. Moreover, PGV-1 and CCA-1.1 suppressed the MMP-9 level in MDA-MB-231 cells. In this study, the molecular mechanisms of the antitumor activity of PGV-1 and CCA-1.1 involve mitotic arrest and ROS production, thus inhibiting cell proliferation. Accordingly, PGV-1 and CCA-1.1 exhibit anticancer properties and should be further developed as prominent antineoplastic drug candidates for breast cancer therapy.

Among breast cancer subtypes, triple-negative breast cancer (TNBC) and human epidermal growth factor receptor 2 (HER2)-positive breast cancer are aggressive types with poor prognosis. Curcumin analogs pentagamavunone-1 (PGV-1) and chemoprevention curcumin analog 1.1 (CCA-1.1) have been explored for anticancer properties against leukemia, breast cancer, and colorectal cancer cells. The present study aimed to evaluate the antitumor activities of PGV-1 and CCA-1.1 in altering the cellular physiology of TNBC and HER2-positive breast cancer cells. This study used MDA-MB-231 (TNBC) and HCC1954 (HER2-positive) cancer cells. The trypan blue exclusion assay was used to determine the cytotoxic effect of PGV-1 and CCA-1.1. Tumor growth inhibition was assessed using a xenograft mice model. The cell cycle profile was examined via flow cytometry, and the mitotic cells were quantified using Hoechst staining. Protein level expression of mitotic kinases (p-cyclin B1, p-PLK1, p-Aurora A, and Aurora A) was determined using western blotting. The intracellular reactive oxygen species (ROS) level was measured via flow cytometry, whereas mitochondrial respiration was monitored with a flux analyzer. The senescence phenomenon was observed using X-gal staining, and the antimigratory activity was monitored using wound healing assay. Then, the secretion level of matrix metalloproteinase 2 and 9 (MMP-2 and MMP9) was examined using gelatin zymography. PGV-1 and CCA-1.1 exhibited cytotoxic activity with GI 50 values of 0.9 and 1.4 µM in MDA-MB-231 cells, respectively. Moreover, they had a cytotoxic effect on HCC1954 cells with GI 50 values of 0.4 and 1.2 µM, respectively. PGV-1 and CCA-1.1 suppressed cell growth with irreversible effects. Oral administration of PGV-1 suppressed tumor growth in mice. The cell cycle profile indicated that PGV-1 induced prometaphase arrest and CCA-1.1 promoted metaphase arrest. Subsequently, PGV-1 tended to inhibit Aurora A kinase in MDA-MB-231 cells and induce cyclin B1 and PLK1 phosphorylation in MDA-MB-231 and HCC1954 cells. CCA-1.1 tended to inhibit Aurora A kinase in MDA-MB-231 cells but not in HCC1954 cells and increase PLK1 and cyclin B1 phosphorylation in HCC1954 cells. The ROS level increased after treatment with PGV-1 or CCA-1.1; this activity might impair mitochondrial function and induce senescence. PGV-1 inhibited MDA-MB-231 and HCC1954 cell migration. Moreover, PGV-1 and CCA-1.1 suppressed the MMP-9 level in MDA-MB-231 cells. In this study, the molecular mechanisms of the antitumor activity of PGV-1 and CCA-1.1 involve mitotic arrest and ROS production, thus inhibiting cell proliferation. Accordingly, PGV-1 and CCA-1.1 exhibit anticancer properties and should be further developed as prominent antineoplastic drug candidates for breast cancer therapy.

Kata Kunci : PGV-1, CCA-1.1, TNBC, HER2-positive breast cancer, mitotic arrest, ROS production

  1. S3-2022-451049-abstract.pdf  
  2. S3-2022-451049-bibliography.pdf  
  3. S3-2022-451049-tableofcontent.pdf  
  4. S3-2022-451049-title.pdf