Kalanchoe blossfeldiana Extract Induces Cell Cycle Arrest and Necrosis in Human Cervical Cancer Cells
Justyna Stefanowicz‑Hajduk1★, Magdalena Gucwa1, Adam Hajduk2, J. Renata Ochocka1★ Corresponding author
- 1Department of Biology and Pharmaceutical Botany, Medical University of Gdańsk, Poland.
- 2Department of Internal Medicine, Connective Tissue Diseases and Geriatrics, Medical University of Gdańsk, Gdańsk, Poland.
CORRESPONDENCE
Justyna Stefanowicz‑Hajduk
Department of Biology and Pharmaceutical Botany, Medical University of Gdańsk, Poland.
Received: 08-03-2019; Revised: 16-04-2019.
Volume 15, Issue 66s · pp. S527–S537 · PUBLISHED 28 November 2019 · DOI: 10.4103/pm.pm_86_19
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ABSTRACT
Objective: The objective of the study is to estimate the cytotoxic activity of KB on human cervical carcinoma (HeLa) cells and determine the mode of cell death. Materials and Methods: The cytotoxic activity of ethanolic extract of leaves of KB was tested on HeLa cells using Real Time xCelligence System (RTCA) and 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT) assay. Cell death, the generation of reactive oxygen species (ROS), depolarization of the mitochondria, cell cycle arrest, and the activity of caspases‑3/7/9 were assessed by flow cytometry or a luminometer. The activity of poly (ADP‑ribose) polymerase (PARP), translocation of apoptosis‑inducing factor (AIF) in the cells, and expression of 92 genes associated with apoptosis/necrosis were also estimated. Results: According to the results, the IC50 values of KB extract were 8.28 ± 0.29 and 9.63 ± 1.07 µg/mL in RTCA and MTT assay, respectively. The results showed a significant increase in the generation of ROS and depolarization of the mitochondria in the dead cell population. Caspases‑3/7/9 were not activated during treatment with KB extract. We also found that the pathway of HeLa cell death was induced by tumor necrosis factor‑related apoptosis‑inducing ligand and was independent of the PARP and AIF. The extract also induced cell cycle arrest in S phase. Conclusions: The results of this study indicate that KB extract induces necrosis in HeLa cells and this process is death receptor‑mediated leading to the overproduction of ROS, mitochondrial dysfunction, and cell cycle arrest.
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Stefanowicz‑Hajduk, J., Gucwa, M., Hajduk, A., & Ochocka, J. R. (2019). Kalanchoe blossfeldiana Extract Induces Cell Cycle Arrest and Necrosis in Human Cervical Cancer Cells. Pharmacognosy Magazine, 15(66s), S527–S537. https://doi.org/10.4103/pm.pm_86_19