Limits...
The enriched fraction of Elephantopus scaber Triggers apoptosis and inhibits multi-drug resistance transporters in human epithelial cancer cells.

Beeran AA, Maliyakkal N, Rao CM, Udupa N - Pharmacogn Mag (2015 Apr-Jun)

Bottom Line: Medicinal plants have played an important role in the development of clinically useful anticancer agents.Cell cycle analysis and micronuclei assay were used to assess cell cycle specific pharmacological effects and drug induced genotoxicty.Thus, ES appears to be potential anticancer agent.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Karnataka, India.

ABSTRACT

Background: Medicinal plants have played an important role in the development of clinically useful anticancer agents. Elephantopus scaber (Asteraceae) (ES) is widely used in Indian traditional system of medicine for the treatment of various ailments including cancer.

Objective: To investigate anticancer effects of ES in human epithelial cancer cells.

Materials and methods: Cytotoxicity of ethanolic extract of ES (ES-ET) and its fractions, such as ES Petroleum ether fraction (ES-PET), ES Dichloromethane fraction (ES DCM), n Butyl alcohol fraction (ES-BT), and ES-Rest (ES-R) were assessed in human epithelial cancer cell lines using sulforhodamine B (SRB) assay. Acridine orange/ethidium bromide assay and Hoechst 33342 assays were used to gauge induction of apoptosis. Cell cycle analysis and micronuclei assay were used to assess cell cycle specific pharmacological effects and drug induced genotoxicty. Further, the ability of ES to inhibit multi drug resistant (MDR) transporters (ABC-B1 and ABC-G2) was determined by Rhodamine (Rho) and Mitoxantrone (MXR) efflux assays.

Results: The enriched fraction of ES (ES DCM) possessed dose-dependent potent cytotoxicity in human epithelial cancer cells. Further, treatment of cancer cells (HeLa, A549, MCF-7, and Caco-2) with ES DCM showed hall mark properties of apoptosis (membrane blebbing, nuclear condensation etc.). Similarly, ES DCM caused enhanced sub G0 content and micronuclei formation indicating the induction of apoptosis and drug induced genotoxicity in cancer cells, respectively. Interestingly, ES DCM inhibited MDR transporters (ABC B1 and ABC G2) in cancer cells.

Conclusion: The enriched fraction of ES imparted cytotoxic effects, triggered apoptosis, induced genotoxicity, and inhibited MDR transporters in human epithelial cancer cells. Thus, ES appears to be potential anticancer agent.

No MeSH data available.


Related in: MedlinePlus

MDR-1 (ABC-B1) inhibitory effects of enriched fraction of Elephantopus scaber (ES-DCM) in human epithelial cancer cells. HeLa (a) and Caco-2 (b) cells were incubated with rhodamine 123 (Rho-123) in Dulbecco's modified eagle medium with 2% fetal bovine serum for 30 min at 37°C (accumulation phase), after which excess dye was washed and cells were re-incubated with vehicle control (dimethyl sulfoxide), verapamil, and ES-DCM for 60 min at 37°C (efflux phase). The mean cellular Rho-123 fluorescence in the efflux phase was analyzed by flow cytometry. Error bars represent ± standard error of the mean; n = 3, ***P < 0.001
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4378122&req=5

Figure 6: MDR-1 (ABC-B1) inhibitory effects of enriched fraction of Elephantopus scaber (ES-DCM) in human epithelial cancer cells. HeLa (a) and Caco-2 (b) cells were incubated with rhodamine 123 (Rho-123) in Dulbecco's modified eagle medium with 2% fetal bovine serum for 30 min at 37°C (accumulation phase), after which excess dye was washed and cells were re-incubated with vehicle control (dimethyl sulfoxide), verapamil, and ES-DCM for 60 min at 37°C (efflux phase). The mean cellular Rho-123 fluorescence in the efflux phase was analyzed by flow cytometry. Error bars represent ± standard error of the mean; n = 3, ***P < 0.001

Mentions: Rho-123 efflux assay revealed that, the MFI of Rho-123 in the efflux phase (with vehicle control, DMSO) was 177.60 ± 7.93 (in HeLa cells). However, in the presence of Ver, the MFI increased to 833.90 ± 17.10, indicating an increased intracellular accumulation of Rho-123 owing to the inhibition of MDR-1 mediated drug efflux [Figure 5a]. Similar to Ver, the treatment with ES-DCM also increased the MFI to 497.60 ± 13.65 [Figure 5a] suggesting that it may function to inhibit MDR-1 mediated drug efflux. Similar result was obtained with Caco-2 cells [Figure 5b]. Compared to Ver (Relative inhibition), ES-DCM contained ~60% and ~67%, inhibition of MDR-1 (ABC-B1) activity in HeLa and Caco-2, respectively [Figure 5a and b]. Similarly, MXR efflux assay revealed that the MFI of MXR in the efflux phase (with vehicle control) was 46.29 ± 1.88 (in A549 cells). However, in the presence of FTC, the MFI enhanced to 91.52 ± 3.38, indicating an enhanced intracellular accumulation of MXR due to the inhibition of BCRP mediated drug efflux [Figure 6a]. Similar to FTC, treatment with ES-DCM enhanced the MFI to 75.76 ± 3.29 suggesting that it may also function to inhibit BCRP activity [Figure 6a]. Similar data was obtained with MCF-7 cells [Figure 6b]. Compared to FTC (Relative inhibition), ES-DCM possessed ~82% and ~78% inhibition of BCRP (ABC-G2) in A549 and MCF-7, respectively [Figure 6a and b]. Thus, these data suggested that the enriched fraction of ES possessed MDR modulating properties, particularly by inhibiting MDR-1 (ABC-B1) and BCRP (ABC-G2) transporters.


The enriched fraction of Elephantopus scaber Triggers apoptosis and inhibits multi-drug resistance transporters in human epithelial cancer cells.

Beeran AA, Maliyakkal N, Rao CM, Udupa N - Pharmacogn Mag (2015 Apr-Jun)

MDR-1 (ABC-B1) inhibitory effects of enriched fraction of Elephantopus scaber (ES-DCM) in human epithelial cancer cells. HeLa (a) and Caco-2 (b) cells were incubated with rhodamine 123 (Rho-123) in Dulbecco's modified eagle medium with 2% fetal bovine serum for 30 min at 37°C (accumulation phase), after which excess dye was washed and cells were re-incubated with vehicle control (dimethyl sulfoxide), verapamil, and ES-DCM for 60 min at 37°C (efflux phase). The mean cellular Rho-123 fluorescence in the efflux phase was analyzed by flow cytometry. Error bars represent ± standard error of the mean; n = 3, ***P < 0.001
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4378122&req=5

Figure 6: MDR-1 (ABC-B1) inhibitory effects of enriched fraction of Elephantopus scaber (ES-DCM) in human epithelial cancer cells. HeLa (a) and Caco-2 (b) cells were incubated with rhodamine 123 (Rho-123) in Dulbecco's modified eagle medium with 2% fetal bovine serum for 30 min at 37°C (accumulation phase), after which excess dye was washed and cells were re-incubated with vehicle control (dimethyl sulfoxide), verapamil, and ES-DCM for 60 min at 37°C (efflux phase). The mean cellular Rho-123 fluorescence in the efflux phase was analyzed by flow cytometry. Error bars represent ± standard error of the mean; n = 3, ***P < 0.001
Mentions: Rho-123 efflux assay revealed that, the MFI of Rho-123 in the efflux phase (with vehicle control, DMSO) was 177.60 ± 7.93 (in HeLa cells). However, in the presence of Ver, the MFI increased to 833.90 ± 17.10, indicating an increased intracellular accumulation of Rho-123 owing to the inhibition of MDR-1 mediated drug efflux [Figure 5a]. Similar to Ver, the treatment with ES-DCM also increased the MFI to 497.60 ± 13.65 [Figure 5a] suggesting that it may function to inhibit MDR-1 mediated drug efflux. Similar result was obtained with Caco-2 cells [Figure 5b]. Compared to Ver (Relative inhibition), ES-DCM contained ~60% and ~67%, inhibition of MDR-1 (ABC-B1) activity in HeLa and Caco-2, respectively [Figure 5a and b]. Similarly, MXR efflux assay revealed that the MFI of MXR in the efflux phase (with vehicle control) was 46.29 ± 1.88 (in A549 cells). However, in the presence of FTC, the MFI enhanced to 91.52 ± 3.38, indicating an enhanced intracellular accumulation of MXR due to the inhibition of BCRP mediated drug efflux [Figure 6a]. Similar to FTC, treatment with ES-DCM enhanced the MFI to 75.76 ± 3.29 suggesting that it may also function to inhibit BCRP activity [Figure 6a]. Similar data was obtained with MCF-7 cells [Figure 6b]. Compared to FTC (Relative inhibition), ES-DCM possessed ~82% and ~78% inhibition of BCRP (ABC-G2) in A549 and MCF-7, respectively [Figure 6a and b]. Thus, these data suggested that the enriched fraction of ES possessed MDR modulating properties, particularly by inhibiting MDR-1 (ABC-B1) and BCRP (ABC-G2) transporters.

Bottom Line: Medicinal plants have played an important role in the development of clinically useful anticancer agents.Cell cycle analysis and micronuclei assay were used to assess cell cycle specific pharmacological effects and drug induced genotoxicty.Thus, ES appears to be potential anticancer agent.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Karnataka, India.

ABSTRACT

Background: Medicinal plants have played an important role in the development of clinically useful anticancer agents. Elephantopus scaber (Asteraceae) (ES) is widely used in Indian traditional system of medicine for the treatment of various ailments including cancer.

Objective: To investigate anticancer effects of ES in human epithelial cancer cells.

Materials and methods: Cytotoxicity of ethanolic extract of ES (ES-ET) and its fractions, such as ES Petroleum ether fraction (ES-PET), ES Dichloromethane fraction (ES DCM), n Butyl alcohol fraction (ES-BT), and ES-Rest (ES-R) were assessed in human epithelial cancer cell lines using sulforhodamine B (SRB) assay. Acridine orange/ethidium bromide assay and Hoechst 33342 assays were used to gauge induction of apoptosis. Cell cycle analysis and micronuclei assay were used to assess cell cycle specific pharmacological effects and drug induced genotoxicty. Further, the ability of ES to inhibit multi drug resistant (MDR) transporters (ABC-B1 and ABC-G2) was determined by Rhodamine (Rho) and Mitoxantrone (MXR) efflux assays.

Results: The enriched fraction of ES (ES DCM) possessed dose-dependent potent cytotoxicity in human epithelial cancer cells. Further, treatment of cancer cells (HeLa, A549, MCF-7, and Caco-2) with ES DCM showed hall mark properties of apoptosis (membrane blebbing, nuclear condensation etc.). Similarly, ES DCM caused enhanced sub G0 content and micronuclei formation indicating the induction of apoptosis and drug induced genotoxicity in cancer cells, respectively. Interestingly, ES DCM inhibited MDR transporters (ABC B1 and ABC G2) in cancer cells.

Conclusion: The enriched fraction of ES imparted cytotoxic effects, triggered apoptosis, induced genotoxicity, and inhibited MDR transporters in human epithelial cancer cells. Thus, ES appears to be potential anticancer agent.

No MeSH data available.


Related in: MedlinePlus