Limits...
Anticancer and apoptotic activities of oleanolic acid are mediated through cell cycle arrest and disruption of mitochondrial membrane potential in HepG2 human hepatocellular carcinoma cells.

Zhu YY, Huang HY, Wu YL - Mol Med Rep (2015)

Bottom Line: Following acridine orange and ethidium bromide staining, treatment with various doses (0, 5, 25 and 50 µM) of oleanolic acid induced typical morphological changes associated with apoptosis, including cell shrinkage, membrane blebbing, nuclear condensation and apoptotic body formation.Oleanolic acid treatment also resulted in fragmentation of nuclear DNA in a dose‑dependent manner, producing the typical features of DNA laddering on an agarose gel.The results also demonstrated that oleanolic acid treatment resulted in a potent loss of mitochondrial membrane potential, which also occurred in a dose‑dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Liver Center, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China.

ABSTRACT
Hepatocellular carcinoma (HCC) is an aggressive form of cancer, with high rates of morbidity and mortality, a poor prognosis and limited therapeutic options. The objective of the present study was to demonstrate the anticancer activity of oleanolic acid in HepG2 human HCC cells. Cell viability was evaluated using an MTT assay, following administration of various doses of oleanolic acid. The effect of oleanolic acid on cell cycle phase distribution and mitochondrial membrane potential was evaluated using flow cytometry with propidium iodide and rhodamine‑123 DNA‑binding cationic fluorescent dyes. Fluorescence microscopy was employed to detect morphological changes in HepG2 cells following oleanolic acid treatment. The results revealed that oleanolic acid induced a dose‑dependent, as well as time‑dependent inhibition in the growth of HepG2 cancer cells. Following acridine orange and ethidium bromide staining, treatment with various doses (0, 5, 25 and 50 µM) of oleanolic acid induced typical morphological changes associated with apoptosis, including cell shrinkage, membrane blebbing, nuclear condensation and apoptotic body formation. Cell cycle analysis revealed that oleanolic acid induced cell cycle arrest in HepG2 cells at the sub‑G1 (apoptotic) phase of the cell cycle, in a dose‑dependent manner. Staining with Annexin V‑fluorescein isothiocyanate and propidium iodide revealed that apoptosis occurred early in these cells. Oleanolic acid treatment also resulted in fragmentation of nuclear DNA in a dose‑dependent manner, producing the typical features of DNA laddering on an agarose gel. The results also demonstrated that oleanolic acid treatment resulted in a potent loss of mitochondrial membrane potential, which also occurred in a dose‑dependent manner. Therefore, oleanolic acid may be used as a therapeutic agent in the treatment of human HCC.

Show MeSH

Related in: MedlinePlus

Analysis of morphological changes in oleanolic acid-treated HepG2 tumor cells. (A) Untreated HepG2 control cells. (B–D) HepG2 cells treated with 5, 25 and 50 µM oleanolic acid, respectively. Cells were exposed to various concentrations of oleanolic acid and morphological changes were observed following 48 h of treatment. Images were captured using an inverted microscope (magnification, ×20).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4581805&req=5

f5-mmr-12-04-5012: Analysis of morphological changes in oleanolic acid-treated HepG2 tumor cells. (A) Untreated HepG2 control cells. (B–D) HepG2 cells treated with 5, 25 and 50 µM oleanolic acid, respectively. Cells were exposed to various concentrations of oleanolic acid and morphological changes were observed following 48 h of treatment. Images were captured using an inverted microscope (magnification, ×20).

Mentions: Apoptosis is a highly organized biochemical process, which eradicates injured or abnormal cells in multicellular organisms. In order to establish whether cell death induced by oleanolic acid is mediated through apoptosis, HepG2 cells were treated with several concentrations of oleanolic acid (0, 5, 25 and 50 µM) for 48 h, and the characteristic morphological features of apoptosis were examined under an inverted light fluorescence microscope. As shown in Fig. 5, compared with viable cells, oleanolic acid treatment at 5 and 25 µM resulted in the appearance of cell shrinkage along with membrane blebbing, which are characteristic features of cell apoptosis. When treated with 50 µM oleanolic acid, almost all the HepG2 cancer cells shrank considerably and no cells with normal morphological features were observed.


Anticancer and apoptotic activities of oleanolic acid are mediated through cell cycle arrest and disruption of mitochondrial membrane potential in HepG2 human hepatocellular carcinoma cells.

Zhu YY, Huang HY, Wu YL - Mol Med Rep (2015)

Analysis of morphological changes in oleanolic acid-treated HepG2 tumor cells. (A) Untreated HepG2 control cells. (B–D) HepG2 cells treated with 5, 25 and 50 µM oleanolic acid, respectively. Cells were exposed to various concentrations of oleanolic acid and morphological changes were observed following 48 h of treatment. Images were captured using an inverted microscope (magnification, ×20).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581805&req=5

f5-mmr-12-04-5012: Analysis of morphological changes in oleanolic acid-treated HepG2 tumor cells. (A) Untreated HepG2 control cells. (B–D) HepG2 cells treated with 5, 25 and 50 µM oleanolic acid, respectively. Cells were exposed to various concentrations of oleanolic acid and morphological changes were observed following 48 h of treatment. Images were captured using an inverted microscope (magnification, ×20).
Mentions: Apoptosis is a highly organized biochemical process, which eradicates injured or abnormal cells in multicellular organisms. In order to establish whether cell death induced by oleanolic acid is mediated through apoptosis, HepG2 cells were treated with several concentrations of oleanolic acid (0, 5, 25 and 50 µM) for 48 h, and the characteristic morphological features of apoptosis were examined under an inverted light fluorescence microscope. As shown in Fig. 5, compared with viable cells, oleanolic acid treatment at 5 and 25 µM resulted in the appearance of cell shrinkage along with membrane blebbing, which are characteristic features of cell apoptosis. When treated with 50 µM oleanolic acid, almost all the HepG2 cancer cells shrank considerably and no cells with normal morphological features were observed.

Bottom Line: Following acridine orange and ethidium bromide staining, treatment with various doses (0, 5, 25 and 50 µM) of oleanolic acid induced typical morphological changes associated with apoptosis, including cell shrinkage, membrane blebbing, nuclear condensation and apoptotic body formation.Oleanolic acid treatment also resulted in fragmentation of nuclear DNA in a dose‑dependent manner, producing the typical features of DNA laddering on an agarose gel.The results also demonstrated that oleanolic acid treatment resulted in a potent loss of mitochondrial membrane potential, which also occurred in a dose‑dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Liver Center, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China.

ABSTRACT
Hepatocellular carcinoma (HCC) is an aggressive form of cancer, with high rates of morbidity and mortality, a poor prognosis and limited therapeutic options. The objective of the present study was to demonstrate the anticancer activity of oleanolic acid in HepG2 human HCC cells. Cell viability was evaluated using an MTT assay, following administration of various doses of oleanolic acid. The effect of oleanolic acid on cell cycle phase distribution and mitochondrial membrane potential was evaluated using flow cytometry with propidium iodide and rhodamine‑123 DNA‑binding cationic fluorescent dyes. Fluorescence microscopy was employed to detect morphological changes in HepG2 cells following oleanolic acid treatment. The results revealed that oleanolic acid induced a dose‑dependent, as well as time‑dependent inhibition in the growth of HepG2 cancer cells. Following acridine orange and ethidium bromide staining, treatment with various doses (0, 5, 25 and 50 µM) of oleanolic acid induced typical morphological changes associated with apoptosis, including cell shrinkage, membrane blebbing, nuclear condensation and apoptotic body formation. Cell cycle analysis revealed that oleanolic acid induced cell cycle arrest in HepG2 cells at the sub‑G1 (apoptotic) phase of the cell cycle, in a dose‑dependent manner. Staining with Annexin V‑fluorescein isothiocyanate and propidium iodide revealed that apoptosis occurred early in these cells. Oleanolic acid treatment also resulted in fragmentation of nuclear DNA in a dose‑dependent manner, producing the typical features of DNA laddering on an agarose gel. The results also demonstrated that oleanolic acid treatment resulted in a potent loss of mitochondrial membrane potential, which also occurred in a dose‑dependent manner. Therefore, oleanolic acid may be used as a therapeutic agent in the treatment of human HCC.

Show MeSH
Related in: MedlinePlus