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Polyphenol-rich extract of Salvia chinensis exhibits anticancer activity in different cancer cell lines, and induces cell cycle arrest at the G₀/G₁-phase, apoptosis and loss of mitochondrial membrane potential in pancreatic cancer cells.

Zhao Q, Huo XC, Sun FD, Dong RQ - Mol Med Rep (2015)

Bottom Line: PC is the fourth most common cause of cancer‑associated mortality in the western world.At present, there is almost no effective treatment available for the treatment of PC.In addition, treatment with the extract induced a significant and concentration-dependent reduction in the ΛΨm of the PC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Yantai Yuhuangding Hospital, Yantai, Shandong 264000, P.R. China.

ABSTRACT
Pancreatic cancer (PC) is one of the most aggressive types of human malignancy, which has an overall 5-year survival rate of <2%. PC is the fourth most common cause of cancer‑associated mortality in the western world. At present, there is almost no effective treatment available for the treatment of PC. The aim of the present study was to evaluate the anticancer potential of a polyphenol enriched extract obtained from Salvia chinensis, a Chinese medicinal plant. An MTT assay was used to evaluate the cell viability of five cancer cell lines and one normal cell line. In addition, the effects of the extract on apoptotic induction, cell cycle phase distribution, DNA damage and loss of mitochondrial membrane potential (ΛΨm) were evaluated in MiapaCa‑2 human PC cells. The effects of the extract on cell cycle phase distribution and ΛΨm were assessed by flow cytometry, using propidium iodide and rhodamine‑123 DNA‑binding fluorescent dyes, respectively. Fluorescence microscopy, using 4',6‑diamidino‑2‑phenylindole as a staining agent, was performed in order to detect the morphological changes of the MiapaCa‑2 cancer cells and the presence of apoptotic bodies following treatment with the extract. The results of the present study demonstrated that the polyphenol‑rich extract from S. chinensis induced potent cytotoxicity in the MCF‑7 human breast cancer cells, A549 human lung cancer cells, HCT‑116 and COLO 205 human colon cancer cells, and MiapaCa‑2 human PC cells. The Colo 205 and MCF‑7 cancer cell lines were the most susceptible to treatment with the extract, which exhibited increased rate of growth inhibition. Fluorescence microscopy revealed characteristic morphological features of apoptosis and detected the appearance of apoptotic bodies following treatment with the extract in the PC cells. Flow cytometric analysis demonstrated that the extract induced G0/G1 cell cycle arrest in a dose‑dependent manner. In addition, treatment with the extract induced a significant and concentration-dependent reduction in the ΛΨm of the PC cells.

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(A Polyphenol-rich extract of Salvia chinensis stimulates apoptosis in MiapaCa-2 human pancreatic cancer cells. (A) Apoptotic cells were evaluated using flow cytometry following annexin V-FITC and PI staining. (a) Control, (b) 20 µg/ml, (c) 40 µg/ml, (d) 60 µg/ml and (e) 80 µg/ml concentration of extract. Cells in the lower left quadrant (annexin V-FITC-/PI-) are viable, those in the lower right quadrant are early apoptotic and those in the upper right quadrant are late apoptotic or necrotic. (B) Percentages of apoptotic cells following treatment with various concentrations of the extract. Data are expressed as the mean + standard deviation of three independent experiments. *P<0.05, **P<0.01, vs 0 µg/ml (control). FITC, fluorescein isothiocyanate; PI, propidium iodide.
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f4-mmr-12-04-4843: (A Polyphenol-rich extract of Salvia chinensis stimulates apoptosis in MiapaCa-2 human pancreatic cancer cells. (A) Apoptotic cells were evaluated using flow cytometry following annexin V-FITC and PI staining. (a) Control, (b) 20 µg/ml, (c) 40 µg/ml, (d) 60 µg/ml and (e) 80 µg/ml concentration of extract. Cells in the lower left quadrant (annexin V-FITC-/PI-) are viable, those in the lower right quadrant are early apoptotic and those in the upper right quadrant are late apoptotic or necrotic. (B) Percentages of apoptotic cells following treatment with various concentrations of the extract. Data are expressed as the mean + standard deviation of three independent experiments. *P<0.05, **P<0.01, vs 0 µg/ml (control). FITC, fluorescein isothiocyanate; PI, propidium iodide.

Mentions: The translocation of phosphatidylserine to the exterior surfaces of the plasma membrane is a distinguishing feature of early apoptosis, which can be identified and detected by annexin V-FITC binding. If cell death occurs, fragmented and damaged DNA becomes permeable for binding with PI (34). Following staining of cells with annexin V in combination with PI, this reagent enters the cells only when the plasma cell membrane has deteriorated. In the present study, flow cytometry revealed that, in the extract-treated cells, a higher number of annexin V-positive cells were detected, compared with the untreated control cells (Fig. 4A and B). The percentage of apoptotic cells was low following treatment with lower concentrations of the extract. However, at higher extract concentrations (60 and 80 µg/ml), the total number of apoptotic cells increased considerably. This assay provided a quantitative estimation of the rate of apoptotic cell death following drug exposure.


Polyphenol-rich extract of Salvia chinensis exhibits anticancer activity in different cancer cell lines, and induces cell cycle arrest at the G₀/G₁-phase, apoptosis and loss of mitochondrial membrane potential in pancreatic cancer cells.

Zhao Q, Huo XC, Sun FD, Dong RQ - Mol Med Rep (2015)

(A Polyphenol-rich extract of Salvia chinensis stimulates apoptosis in MiapaCa-2 human pancreatic cancer cells. (A) Apoptotic cells were evaluated using flow cytometry following annexin V-FITC and PI staining. (a) Control, (b) 20 µg/ml, (c) 40 µg/ml, (d) 60 µg/ml and (e) 80 µg/ml concentration of extract. Cells in the lower left quadrant (annexin V-FITC-/PI-) are viable, those in the lower right quadrant are early apoptotic and those in the upper right quadrant are late apoptotic or necrotic. (B) Percentages of apoptotic cells following treatment with various concentrations of the extract. Data are expressed as the mean + standard deviation of three independent experiments. *P<0.05, **P<0.01, vs 0 µg/ml (control). FITC, fluorescein isothiocyanate; PI, propidium iodide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581742&req=5

f4-mmr-12-04-4843: (A Polyphenol-rich extract of Salvia chinensis stimulates apoptosis in MiapaCa-2 human pancreatic cancer cells. (A) Apoptotic cells were evaluated using flow cytometry following annexin V-FITC and PI staining. (a) Control, (b) 20 µg/ml, (c) 40 µg/ml, (d) 60 µg/ml and (e) 80 µg/ml concentration of extract. Cells in the lower left quadrant (annexin V-FITC-/PI-) are viable, those in the lower right quadrant are early apoptotic and those in the upper right quadrant are late apoptotic or necrotic. (B) Percentages of apoptotic cells following treatment with various concentrations of the extract. Data are expressed as the mean + standard deviation of three independent experiments. *P<0.05, **P<0.01, vs 0 µg/ml (control). FITC, fluorescein isothiocyanate; PI, propidium iodide.
Mentions: The translocation of phosphatidylserine to the exterior surfaces of the plasma membrane is a distinguishing feature of early apoptosis, which can be identified and detected by annexin V-FITC binding. If cell death occurs, fragmented and damaged DNA becomes permeable for binding with PI (34). Following staining of cells with annexin V in combination with PI, this reagent enters the cells only when the plasma cell membrane has deteriorated. In the present study, flow cytometry revealed that, in the extract-treated cells, a higher number of annexin V-positive cells were detected, compared with the untreated control cells (Fig. 4A and B). The percentage of apoptotic cells was low following treatment with lower concentrations of the extract. However, at higher extract concentrations (60 and 80 µg/ml), the total number of apoptotic cells increased considerably. This assay provided a quantitative estimation of the rate of apoptotic cell death following drug exposure.

Bottom Line: PC is the fourth most common cause of cancer‑associated mortality in the western world.At present, there is almost no effective treatment available for the treatment of PC.In addition, treatment with the extract induced a significant and concentration-dependent reduction in the ΛΨm of the PC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Yantai Yuhuangding Hospital, Yantai, Shandong 264000, P.R. China.

ABSTRACT
Pancreatic cancer (PC) is one of the most aggressive types of human malignancy, which has an overall 5-year survival rate of <2%. PC is the fourth most common cause of cancer‑associated mortality in the western world. At present, there is almost no effective treatment available for the treatment of PC. The aim of the present study was to evaluate the anticancer potential of a polyphenol enriched extract obtained from Salvia chinensis, a Chinese medicinal plant. An MTT assay was used to evaluate the cell viability of five cancer cell lines and one normal cell line. In addition, the effects of the extract on apoptotic induction, cell cycle phase distribution, DNA damage and loss of mitochondrial membrane potential (ΛΨm) were evaluated in MiapaCa‑2 human PC cells. The effects of the extract on cell cycle phase distribution and ΛΨm were assessed by flow cytometry, using propidium iodide and rhodamine‑123 DNA‑binding fluorescent dyes, respectively. Fluorescence microscopy, using 4',6‑diamidino‑2‑phenylindole as a staining agent, was performed in order to detect the morphological changes of the MiapaCa‑2 cancer cells and the presence of apoptotic bodies following treatment with the extract. The results of the present study demonstrated that the polyphenol‑rich extract from S. chinensis induced potent cytotoxicity in the MCF‑7 human breast cancer cells, A549 human lung cancer cells, HCT‑116 and COLO 205 human colon cancer cells, and MiapaCa‑2 human PC cells. The Colo 205 and MCF‑7 cancer cell lines were the most susceptible to treatment with the extract, which exhibited increased rate of growth inhibition. Fluorescence microscopy revealed characteristic morphological features of apoptosis and detected the appearance of apoptotic bodies following treatment with the extract in the PC cells. Flow cytometric analysis demonstrated that the extract induced G0/G1 cell cycle arrest in a dose‑dependent manner. In addition, treatment with the extract induced a significant and concentration-dependent reduction in the ΛΨm of the PC cells.

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Related in: MedlinePlus