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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) The influence of PCAE on apoptosis/necrosis in Ishikawa cells; (B) Total apoptosis in Ishikawa cells following incubation with PCAE for 4 h. (*p < 0.05 vs. PCAE 0 mg/mL control group). Influence of PCAE on cell cycle progression/distribution in Ishikawa cells: (C) cell cycle analysis of Ishikawa cells after being cultured with PCAE for 24 h; and (D) PCAE induced an increase in sub G1 (%). Cells underwent dual staining using propidium iodide to analyze DNA content, which was then quantified using flow cytometry. The * symbol in each group of bars indicates that the number of sub G1 cells in the PCAE treatment group was significantly greater than that of the control at a significance level of p < 0.05.
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ijms-16-12424-f002: (A) The influence of PCAE on apoptosis/necrosis in Ishikawa cells; (B) Total apoptosis in Ishikawa cells following incubation with PCAE for 4 h. (*p < 0.05 vs. PCAE 0 mg/mL control group). Influence of PCAE on cell cycle progression/distribution in Ishikawa cells: (C) cell cycle analysis of Ishikawa cells after being cultured with PCAE for 24 h; and (D) PCAE induced an increase in sub G1 (%). Cells underwent dual staining using propidium iodide to analyze DNA content, which was then quantified using flow cytometry. The * symbol in each group of bars indicates that the number of sub G1 cells in the PCAE treatment group was significantly greater than that of the control at a significance level of p < 0.05.

Mentions: The ApopNexin FITC apoptosis detection kit was used to identify apoptotic Ishikawa cells following exposure to PCAE for 6 h. PI-annexin-V double staining was used to differentiate intact cells from early apoptotic cells, late apoptotic cells, and dead (necrotic) cells as well as to investigate apoptosis in greater detail. Typical assay results are presented in Figure 2A, in which annexin V-FITC deposits indicate the existence of apoptotic cells. As shown in Figure 2B, apoptosis was induced in a dose-dependent manner when administered at 2 and 4 mg/mL (y = 19.72 x − 9.33 R2 = 0.9265). Increases of the percentages of apoptotic Ishikawa cells was observed at all dose levels following treatment for 6 h, wherein approximately 16.17% ± 2.02% of the Ishikawa cells were in early or late stages of apoptosis. The percentage of apoptotic Ishikawa cells increased to 19.73% ± 2.46% under PCAE treatment of 1 mg/mL. When the PCAE concentration was increased to 2 and 4 mg/mL, the percentage apoptotic Ishikawa cells increased to 53.25% ± 6.66% and 70.73% ± 8.84%, respectively. Taken together, these observations strongly imply that PCAE significantly elevated apoptosis in EC 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) The influence of PCAE on apoptosis/necrosis in Ishikawa cells; (B) Total apoptosis in Ishikawa cells following incubation with PCAE for 4 h. (*p < 0.05 vs. PCAE 0 mg/mL control group). Influence of PCAE on cell cycle progression/distribution in Ishikawa cells: (C) cell cycle analysis of Ishikawa cells after being cultured with PCAE for 24 h; and (D) PCAE induced an increase in sub G1 (%). Cells underwent dual staining using propidium iodide to analyze DNA content, which was then quantified using flow cytometry. The * symbol in each group of bars indicates that the number of sub G1 cells in the PCAE treatment group was significantly greater than that of the control at a significance level of p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-12424-f002: (A) The influence of PCAE on apoptosis/necrosis in Ishikawa cells; (B) Total apoptosis in Ishikawa cells following incubation with PCAE for 4 h. (*p < 0.05 vs. PCAE 0 mg/mL control group). Influence of PCAE on cell cycle progression/distribution in Ishikawa cells: (C) cell cycle analysis of Ishikawa cells after being cultured with PCAE for 24 h; and (D) PCAE induced an increase in sub G1 (%). Cells underwent dual staining using propidium iodide to analyze DNA content, which was then quantified using flow cytometry. The * symbol in each group of bars indicates that the number of sub G1 cells in the PCAE treatment group was significantly greater than that of the control at a significance level of p < 0.05.
Mentions: The ApopNexin FITC apoptosis detection kit was used to identify apoptotic Ishikawa cells following exposure to PCAE for 6 h. PI-annexin-V double staining was used to differentiate intact cells from early apoptotic cells, late apoptotic cells, and dead (necrotic) cells as well as to investigate apoptosis in greater detail. Typical assay results are presented in Figure 2A, in which annexin V-FITC deposits indicate the existence of apoptotic cells. As shown in Figure 2B, apoptosis was induced in a dose-dependent manner when administered at 2 and 4 mg/mL (y = 19.72 x − 9.33 R2 = 0.9265). Increases of the percentages of apoptotic Ishikawa cells was observed at all dose levels following treatment for 6 h, wherein approximately 16.17% ± 2.02% of the Ishikawa cells were in early or late stages of apoptosis. The percentage of apoptotic Ishikawa cells increased to 19.73% ± 2.46% under PCAE treatment of 1 mg/mL. When the PCAE concentration was increased to 2 and 4 mg/mL, the percentage apoptotic Ishikawa cells increased to 53.25% ± 6.66% and 70.73% ± 8.84%, respectively. Taken together, these observations strongly imply that PCAE significantly elevated apoptosis in EC 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