Morphological and biochemical effects of carnosic acid on human hepatocellular carcinoma HepG2 cells
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Abstract
Antecedentes / objetivo : La muerte celular autofágica y la apoptosis de células tumorales se ha convertido en uno de los principales objetivos en el tratamiento del cáncer, mientras que las líneas celulares tumorales se utilizan principalmente en estudios para proporcionar datos importantes para la evaluación de posibles sustancias anticancerígenas. En este estudio, nuestro objetivo fue evaluar los cambios morfológicos y bioquímicos, incluida la tasa de apoptosis y los niveles de alfa fetoproteína (AFP) a diferentes concentraciones de ácido carnósico (CA) en células de carcinoma hepatocelular humano HepG2. Materiales y métodos : Carcinoma hepatocelular humano (células HepG2 de séptimo pase). Las líneas celulares se cultivaron en cubreobjetos de vidrio Schott D263M de 11 µM colocados en placas de 12 pocillos y se trataron con DMSO, concentraciones de CA 1, 2,5, 5 y 10 µM durante 24 horas. 48 y 72 horas. Los datos morfológicos y bioquímicos se registraron diariamente, incluidas las tasas de apoptosis demostradas por Caspasa 3, las expresiones de Anexina V bajo luz invertida y microscopía de inmunofluorescencia, luego se analizaron los datos para determinar la significación estadística. Los niveles de AFP, albúmina y ganancias totales se analizaron espectrofotométricamente para evaluación bioquímica. Resultados : Nuestros resultados muestran que CA inhibió significativamente la proliferación de células HepG2 de una manera dependiente de la dosis y el tiempo y causó significativamente la formación de vacuolas autofágicas comenzando desde 5 µM y alcanzando significancia a concentraciones de 10 µM. Se produce una disminución significativa en la AFP cuando se examinan las expresiones de 48 y 72 horas, alcanzando el nivel más bajo a las 72 horas en el grupo de CA 10 µM. Además, el aumento en los niveles de albúmina alcanzó la significación solo en el grupo de 48 h, mientras que también se observaron aumentos no significativos en los grupos de 24 hy 72 h. Conclusión: Nuestro estudio actual demuestra un aumento significativo en las tasas de apoptosis por el ácido carnósico principalmente a concentraciones de 10 µM, lo que respalda su efecto anticancerígeno en las células HepG2. Estos hallazgos también están respaldados por cambios en los análisis bioquímicos de los niveles de albúmina y AFP a concentraciones de 10 µM.
References
ADEN, D. P., FOGEL, A., PLOTKIN, S., DAMJANOV, I. & KNOWLES, B. B. 1979. Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line. Nature, 282, 615-616. https://doi.org/10.1038/282615a0
AKıN, U. N., BAKıR, E., ÖKÇESIZ, A. & EKEN, A. 2019. Evaluation of Cytotoxic Effects of Carnosic Acid Alone and Combination with Cisplatin in HepG2 Cells. Multidisciplinary Digital Publishing Institute Proceedings, 40, 39. https://doi.org/10.3390/proceedings2019040039
ALEXANDROV, K., ROJAS, M. & ROLANDO, C. 2006. DNA damage by benzo (a) pyrene in human cells is increased by cigarette smoke and decreased by a filter containing rosemary extract, which lowers free radicals. Cancer research, 66, 11938-11945. https://doi.org/10.1158/0008-5472.can-06-3277
ALTIN, J. G. & SLOAN, E. K. 1997. The role of CD45 and CD45-associated molecules in T cell activation. Immunol Cell Biol, 75, 430-45. https://doi.org/10.1038/icb.1997.68
BAHRI, S., MIES, F., BEN ALI, R., MLIKA, M., JAMELEDDINE, S., MC ENTEE, K. & SHLYONSKY, V. 2017. Rosmarinic acid potentiates carnosic acid induced apoptosis in lung fibroblasts. PloS one, 12, e0184368. https://doi.org/10.1371/journal.pone.0184368
BORRÁS-LINARES, I., PÉREZ-SÁNCHEZ, A., LOZANO-SÁNCHEZ, J., BARRAJÓN-CATALÁN, E., ARRÁEZ-ROMÁN, D., CIFUENTES, A., MICOL, V. & CARRETERO, A. S. 2015. A bioguided identification of the active compounds that contribute to the antiproliferative/cytotoxic effects of rosemary extract on colon cancer cells. Food and Chemical Toxicology, 80, 215-222. https://doi.org/10.1016/j.fct.2015.03.013
BOSCH, F. X., RIBES, J., DÍAZ, M. & CLÉRIES, R. 2004. Primary liver cancer: worldwide incidence and trends. Gastroenterology, 127, S5-S16. https://doi.org/10.1053/j.gastro.2004.09.011
CHEUNG, S. & TAI, J. 2007. Anti-proliferative and antioxidant properties of rosemary Rosmarinus officinalis. Oncology reports, 17, 1525-1531. https://pubmed.ncbi.nlm.nih.gov/17487414/
CUVELIER, M. E., BERSET, C. & RICHARD, H. 1994. Antioxidant constituents in sage (Salvia officinalis). Journal of Agricultural and food chemistry, 42, 665-669. https://pubs.acs.org/doi/10.1021/jf00039a012
CUVELIER, M. E., RICHARD, H. & BERSET, C. 1996. Antioxidative activity and phenolic composition of pilot‐plant and commercial extracts of sage and rosemary. Journal of the American Oil Chemists' Society, 73, 645-652. https://doi.org/10.1007/BF02518121
GAO, Q., LIU, H., YAO, Y., GENG, L., ZHANG, X., JIANG, L., SHI, B. & YANG, F. 2015. Carnosic acid induces autophagic cell death through inhibition of the Akt/mTOR pathway in human hepatoma cells. Journal of Applied Toxicology, 35, 485-492. https://doi.org/10.1002/jat.3049
GONZÁLEZ-VALLINAS, M., REGLERO, G. & RAMÍREZ DE MOLINA, A. 2015. Rosemary (Rosmarinus officinalis L.) extract as a potential complementary agent in anticancer therapy. Nutrition and cancer, 67, 1223-1231. https://doi.org/10.1080/01635581.2015.1082110
HOZAYEN, W., SOLIMAN, H. & DESOUKY, E. M. 2014. Potential protective effects of rosemary extract, against aspartame toxicity in male rats. J Inter Acad Res Multidisc, 2, 111-125.
HUANG, S.-S. & ZHENG, R.-L. 2006. Rosmarinic acid inhibits angiogenesis and its mechanism of action in vitro. Cancer letters, 239, 271-280. https://doi.org/10.1016/j.canlet.2005.08.025
KIM, D.-S., KIM, H.-R., WOO, E.-R., HONG, S.-T., CHAE, H.-J. & CHAE, S.-W. 2005. Inhibitory effects of rosmarinic acid on adriamycin-induced apoptosis in H9c2 cardiac muscle cells by inhibiting reactive oxygen species and the activations of c-Jun N-terminal kinase and extracellular signal-regulated kinase. Biochemical Pharmacology, 70, 1066-1078. https://doi.org/10.1016/j.bcp.2005.06.026
LI, G.-S., JIANG, W.-L., TIAN, J.-W., QU, G.-W., ZHU, H.-B. & FU, F.-H. 2010. In vitro and in vivo antifibrotic effects of rosmarinic acid on experimental liver fibrosis. Phytomedicine, 17, 282-288. https://doi.org/10.1016/j.phymed.2009.05.002
LLOVET, J. M., FUSTER, J. & BRUIX, J. 2004. The Barcelona approach: diagnosis, staging, and treatment of hepatocellular carcinoma. Liver transplantation, 10, S115-S120. https://doi.org/10.1002/lt.20034
MOORE, J., YOUSEF, M. & TSIANI, E. 2016. Anticancer effects of rosemary (Rosmarinus officinalis L.) extract and rosemary extract polyphenols. Nutrients, 8, 731. https://doi.org/10.3390/nu8110731
PETIWALA, S. M. & JOHNSON, J. J. 2015. Diterpenes from rosemary (Rosmarinus officinalis): Defining their potential for anti-cancer activity. Cancer letters, 367, 93-102. https://doi.org/10.1016/j.canlet.2015.07.005
PETIWALA, S. M., PUTHENVEETIL, A. G. & JOHNSON, J. 2013. Polyphenols from the Mediterranean herb rosemary (Rosmarinus officinalis) for prostate cancer. Frontiers in pharmacology, 4, 29. https://doi.org/10.3389/fphar.2013.00029
SHABTAY, A., SHARABANI, H., BARVISH, Z., KAFKA, M., AMICHAY, D., LEVY, J., SHARONI, Y., USKOKOVIC, M. R., STUDZINSKI, G. P. & DANILENKO, M. 2008. Synergistic antileukemic activity of carnosic acid-rich rosemary extract and the 19-nor Gemini vitamin D analogue in a mouse model of systemic acute myeloid leukemia. Oncology, 75, 203-214. https://doi.org/10.1159/000163849
SHAO, N., MAO, J., XUE, L., WANG, R., ZHI, F. & LAN, Q. 2019. Carnosic acid potentiates the anticancer effect of temozolomide by inducing apoptosis and autophagy in glioma. Journal of Neuro-Oncology, 141, 277-288. https://doi.org/10.1007/s11060-018-03043-5
SLAMEŇOVÁ, D., KUBOŠKOVÁ, K. N., HORVÁTHOVÁ, E. & ROBICHOVÁ, S. 2002. Rosemary-stimulated reduction of DNA strand breaks and FPG-sensitive sites in mammalian cells treated with H2O2 or visible light-excited Methylene Blue. Cancer letters, 177, 145-153. https://doi.org/10.1016/s0304-3835(01)00784-4
WILDING, J. L. & BODMER, W. F. 2014. Cancer cell lines for drug discovery and development. Cancer research, 74, 2377-2384. https://doi.org/10.1158/0008-5472.can-13-2971
YESIL-CELIKTAS, O., SEVIMLI, C., BEDIR, E. & VARDAR-SUKAN, F. 2010. Inhibitory effects of rosemary extracts, carnosic acid and rosmarinic acid on the growth of various human cancer cell lines. Plant foods for human nutrition, 65, 158-163. https://doi.org/10.1007/s11130-010-0166-4
YUAN, J. & KROEMER, G. 2010. Alternative cell death mechanisms in development and beyond. Genes & development, 24, 2592-2602. https://doi.org/10.1101/gad.1984410