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Induction of Apoptosis in Endometrial Cancer (Ishikawa) Cells by Pogostemon cablin Aqueous Extract (PCAE).

Tsai CC, Chang YH, Chang CC, Cheng YM, Ou YC, Chien CC, Hsu YC - Int J Mol Sci (2015)

Bottom Line: Gene expression profiling (GEP) results further suggest that, in addition to its known effects with regard to EC prevention, PCAE may also exert antitumor activity on established EC cells.Many previous studies have identified the chemo-preventive effects of natural plant materials and the potential role of these materials in chemotherapy.This current study used human EC Ishikawa cells to investigate the anti-tumor effects of PCAE in EC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Kaohsiung 83301, Taiwan. nick@adm.cgmh.org.tw.

ABSTRACT
Pogostemon cablin (PC) is a traditional herbal medicine used in the treatment of the common cold, nausea, diarrhea, and even for headaches and fever. However, the mechanisms underlying the anti-proliferative activity of PC in endometrial cancer (EC) cells have yet to be fully elucidated. This study investigated the anticancer effects of an aqueous extract of Pogostemon cablin (PCAE), specifically induced apoptosis in EC (Ishikawa) cells. Proliferation of EC cells following exposure to PCAE was assessed by an MTT assay. DNA content and the induction of cell cycle apoptosis were analyzed by flow cytometry (FACS Calibur). Protein caspase-3 and, -9 as well as AIF were investigated using Western blot. Our results demonstrate growth inhibition of Ishikawa cells by PCAE. Furthermore, caspase-3 activity caused PCAE-treated cell lines to accumulate in apoptosis. Gene expression profiling (GEP) results further suggest that, in addition to its known effects with regard to EC prevention, PCAE may also exert antitumor activity on established EC cells. Many previous studies have identified the chemo-preventive effects of natural plant materials and the potential role of these materials in chemotherapy. This current study used human EC Ishikawa cells to investigate the anti-tumor effects of PCAE in EC cells. Our results demonstrate that PCAE inhibits the growth of cancer cells and induces apoptosis, which suggests the potential applicability of PCAE as an antitumor agent.

No MeSH data available.


Related in: MedlinePlus

(A) Reduction of mitochondrial membrane potential (MMP) in Ishikawa cells by PCAE, as determined by JC-1 staining and detected by flow cytometry: MMP was shown to be significantly reduced in Ishikawa cells treated with PCAE (0, 1, 2, and 5 mg/mL); (B) Quantification by flow cytometry; (C) Caspase-3 activity in Ishikawa cells following treatment with PCAE for 24 h. Following treatment, the cells were harvested and labeled using FITC rabbit anti-active caspase-3; (D) Activation was quantified by flow cytometry; and (E) Cells were treated with PCAE for 24 h and procaspase-3, and -9, as well as AIF proteins were subsequently detected by Western blot analysis. All data are reported as the mean (±SEM) of at least three separate experiments. The * symbol in each group of bars indicates significant differences between PCAE treatment and control groups (p < 0.05).
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ijms-16-12424-f003: (A) Reduction of mitochondrial membrane potential (MMP) in Ishikawa cells by PCAE, as determined by JC-1 staining and detected by flow cytometry: MMP was shown to be significantly reduced in Ishikawa cells treated with PCAE (0, 1, 2, and 5 mg/mL); (B) Quantification by flow cytometry; (C) Caspase-3 activity in Ishikawa cells following treatment with PCAE for 24 h. Following treatment, the cells were harvested and labeled using FITC rabbit anti-active caspase-3; (D) Activation was quantified by flow cytometry; and (E) Cells were treated with PCAE for 24 h and procaspase-3, and -9, as well as AIF proteins were subsequently detected by Western blot analysis. All data are reported as the mean (±SEM) of at least three separate experiments. The * symbol in each group of bars indicates significant differences between PCAE treatment and control groups (p < 0.05).

Mentions: The loss of mitochondrial membrane potential is a hallmark of apoptosis, coinciding with caspase activation. In non-apoptotic cells, JC-1 exists as a monomer in the cytosol (green) and accumulates as an aggregate in the mitochondria (red). Figure 3A presents typical FL-1/FL-2 dot plots associated with the JC-1 staining of Ishikawa cells with and without apoptosis. Untreated Ishikawa cells presented no signs of apoptosis and have red fluorescing J-aggregates. The green fluorescing monomers shown in the lower part of the figure indicate apoptotic cells (PCAE treatment using 1, 2, or 4 mg/mL). Figure 3B presents the percentages of apoptotic Ishikawa cells in various PCAE-treated groups as analyzed by flow cytometry. A decrease in mitochondrial membrane potential was observed at all dosage levels following treatment for 6 h. Specifically, treatment with 1 mg/mL PCAE led to a reduction of 2.06 x (y = 191.64 x − 117.23 R2 = 0.9673). A further increase in the concentrations of PCAE to 2 and 4 mg/mL led to reductions of 5.01 x and 6.41 x, respectively. These findings imply that PCAE significantly reduced the mitochondrial membrane potential of Ishikawa cells.


Induction of Apoptosis in Endometrial Cancer (Ishikawa) Cells by Pogostemon cablin Aqueous Extract (PCAE).

Tsai CC, Chang YH, Chang CC, Cheng YM, Ou YC, Chien CC, Hsu YC - Int J Mol Sci (2015)

(A) Reduction of mitochondrial membrane potential (MMP) in Ishikawa cells by PCAE, as determined by JC-1 staining and detected by flow cytometry: MMP was shown to be significantly reduced in Ishikawa cells treated with PCAE (0, 1, 2, and 5 mg/mL); (B) Quantification by flow cytometry; (C) Caspase-3 activity in Ishikawa cells following treatment with PCAE for 24 h. Following treatment, the cells were harvested and labeled using FITC rabbit anti-active caspase-3; (D) Activation was quantified by flow cytometry; and (E) Cells were treated with PCAE for 24 h and procaspase-3, and -9, as well as AIF proteins were subsequently detected by Western blot analysis. All data are reported as the mean (±SEM) of at least three separate experiments. The * symbol in each group of bars indicates significant differences between PCAE treatment and control groups (p < 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490452&req=5

ijms-16-12424-f003: (A) Reduction of mitochondrial membrane potential (MMP) in Ishikawa cells by PCAE, as determined by JC-1 staining and detected by flow cytometry: MMP was shown to be significantly reduced in Ishikawa cells treated with PCAE (0, 1, 2, and 5 mg/mL); (B) Quantification by flow cytometry; (C) Caspase-3 activity in Ishikawa cells following treatment with PCAE for 24 h. Following treatment, the cells were harvested and labeled using FITC rabbit anti-active caspase-3; (D) Activation was quantified by flow cytometry; and (E) Cells were treated with PCAE for 24 h and procaspase-3, and -9, as well as AIF proteins were subsequently detected by Western blot analysis. All data are reported as the mean (±SEM) of at least three separate experiments. The * symbol in each group of bars indicates significant differences between PCAE treatment and control groups (p < 0.05).
Mentions: The loss of mitochondrial membrane potential is a hallmark of apoptosis, coinciding with caspase activation. In non-apoptotic cells, JC-1 exists as a monomer in the cytosol (green) and accumulates as an aggregate in the mitochondria (red). Figure 3A presents typical FL-1/FL-2 dot plots associated with the JC-1 staining of Ishikawa cells with and without apoptosis. Untreated Ishikawa cells presented no signs of apoptosis and have red fluorescing J-aggregates. The green fluorescing monomers shown in the lower part of the figure indicate apoptotic cells (PCAE treatment using 1, 2, or 4 mg/mL). Figure 3B presents the percentages of apoptotic Ishikawa cells in various PCAE-treated groups as analyzed by flow cytometry. A decrease in mitochondrial membrane potential was observed at all dosage levels following treatment for 6 h. Specifically, treatment with 1 mg/mL PCAE led to a reduction of 2.06 x (y = 191.64 x − 117.23 R2 = 0.9673). A further increase in the concentrations of PCAE to 2 and 4 mg/mL led to reductions of 5.01 x and 6.41 x, respectively. These findings imply that PCAE significantly reduced the mitochondrial membrane potential of Ishikawa cells.

Bottom Line: Gene expression profiling (GEP) results further suggest that, in addition to its known effects with regard to EC prevention, PCAE may also exert antitumor activity on established EC cells.Many previous studies have identified the chemo-preventive effects of natural plant materials and the potential role of these materials in chemotherapy.This current study used human EC Ishikawa cells to investigate the anti-tumor effects of PCAE in EC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Kaohsiung 83301, Taiwan. nick@adm.cgmh.org.tw.

ABSTRACT
Pogostemon cablin (PC) is a traditional herbal medicine used in the treatment of the common cold, nausea, diarrhea, and even for headaches and fever. However, the mechanisms underlying the anti-proliferative activity of PC in endometrial cancer (EC) cells have yet to be fully elucidated. This study investigated the anticancer effects of an aqueous extract of Pogostemon cablin (PCAE), specifically induced apoptosis in EC (Ishikawa) cells. Proliferation of EC cells following exposure to PCAE was assessed by an MTT assay. DNA content and the induction of cell cycle apoptosis were analyzed by flow cytometry (FACS Calibur). Protein caspase-3 and, -9 as well as AIF were investigated using Western blot. Our results demonstrate growth inhibition of Ishikawa cells by PCAE. Furthermore, caspase-3 activity caused PCAE-treated cell lines to accumulate in apoptosis. Gene expression profiling (GEP) results further suggest that, in addition to its known effects with regard to EC prevention, PCAE may also exert antitumor activity on established EC cells. Many previous studies have identified the chemo-preventive effects of natural plant materials and the potential role of these materials in chemotherapy. This current study used human EC Ishikawa cells to investigate the anti-tumor effects of PCAE in EC cells. Our results demonstrate that PCAE inhibits the growth of cancer cells and induces apoptosis, which suggests the potential applicability of PCAE as an antitumor agent.

No MeSH data available.


Related in: MedlinePlus