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Ethyl acetate extract from marine sponge Hyattella cribriformis exhibit potent anticancer activity by promoting tubulin polymerization as evidenced mitotic arrest and induction of apoptosis.

Annamalai P, Thayman M, Rajan S, Raman LS, Ramasubbu S, Perumal P - Pharmacogn Mag (2015 Apr-Jun)

Bottom Line: EA fraction exhibited potent inhibition of cancer cell growth and resulted in 50% growth inhibition (GI50) of 0.27 μg/mL in A673 cell line.Fraction induced DNA fragmentation in HeLa cells as evidence of apoptosis.Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Periyar University, Karuppur, Salem, India.

ABSTRACT

Background: Marine sponges are important sources of bioactive compounds.

Objective: This study investigated the anticancer properties of Hyattella cribriformis ethyl acetate (EA) fraction in various cancer and normal cell lines.

Materials and methods: anticancer assay was carried out in 15 cell lines to evaluate the anticancer potential of the EA fraction. Impact on cell cycle distribution was determined using flow cytometry. The fraction was investigated for interfering microtubules assembly in both in vitro and cellular assay. Further studies were conducted to determine the fraction induced cell death (apoptosis) using calcein/propidium iodide dual staining, activated caspase-3 and phosphorylation of Bcl-2 protein at Ser70. DNA fragmentation assay was performed to confirm the apoptosis.

Results: EA fraction exhibited potent inhibition of cancer cell growth and resulted in 50% growth inhibition (GI50) of 0.27 μg/mL in A673 cell line. Sarcoma (MG-63, Saos-2) and ovarian (SK-OV-3 and OVCAR-3) cancer cell lines also showed superior anticancer activity GI50 of 1.0 μg/mL. Colon and breast cancer cell lines exhibited moderate GI compare other cancer cell lines and normal human lung fibroblast showed GI50 of 15.6 μg/mL. EA fraction showed potent G2/M phase arrest in A673 cell line and induced apoptosis at 48 h exposure. EA fraction promoted microtubule polymerization in tubulin polymerization assay and increased level of polymerized tubulin in the HeLa cells. Fraction induced the activation of caspase-3 and phosphorylation of Bcl-2 anti-apoptotic protein. Fraction induced DNA fragmentation in HeLa cells as evidence of apoptosis.

Conclusion: Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.

No MeSH data available.


Related in: MedlinePlus

Flow cytometric analysis of cell cycle distribution on the A673 cells treated 24 and 48 h with different concentration of ethyl acetate fraction from Hyattella cribriformis in comparison with untreated control
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Figure 4: Flow cytometric analysis of cell cycle distribution on the A673 cells treated 24 and 48 h with different concentration of ethyl acetate fraction from Hyattella cribriformis in comparison with untreated control

Mentions: As observed in 24 h, cells exposed for 48 h were also primarily arrested in G2-M phase (31%, 28%, 31% and 28% vs. 23%) with a concomitant decrease in G0-G1 population (52%, 50%, 36% and 27% vs. 58% control). The percentage of cells in S phase remained stable at all doses. Cells exposed to 10 and 3 μg concentration of EA for 48 h alone exhibited an increase in sub G0-G1 population, indicating an induction of apoptosis [Figure 4].


Ethyl acetate extract from marine sponge Hyattella cribriformis exhibit potent anticancer activity by promoting tubulin polymerization as evidenced mitotic arrest and induction of apoptosis.

Annamalai P, Thayman M, Rajan S, Raman LS, Ramasubbu S, Perumal P - Pharmacogn Mag (2015 Apr-Jun)

Flow cytometric analysis of cell cycle distribution on the A673 cells treated 24 and 48 h with different concentration of ethyl acetate fraction from Hyattella cribriformis in comparison with untreated control
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4378133&req=5

Figure 4: Flow cytometric analysis of cell cycle distribution on the A673 cells treated 24 and 48 h with different concentration of ethyl acetate fraction from Hyattella cribriformis in comparison with untreated control
Mentions: As observed in 24 h, cells exposed for 48 h were also primarily arrested in G2-M phase (31%, 28%, 31% and 28% vs. 23%) with a concomitant decrease in G0-G1 population (52%, 50%, 36% and 27% vs. 58% control). The percentage of cells in S phase remained stable at all doses. Cells exposed to 10 and 3 μg concentration of EA for 48 h alone exhibited an increase in sub G0-G1 population, indicating an induction of apoptosis [Figure 4].

Bottom Line: EA fraction exhibited potent inhibition of cancer cell growth and resulted in 50% growth inhibition (GI50) of 0.27 μg/mL in A673 cell line.Fraction induced DNA fragmentation in HeLa cells as evidence of apoptosis.Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Periyar University, Karuppur, Salem, India.

ABSTRACT

Background: Marine sponges are important sources of bioactive compounds.

Objective: This study investigated the anticancer properties of Hyattella cribriformis ethyl acetate (EA) fraction in various cancer and normal cell lines.

Materials and methods: anticancer assay was carried out in 15 cell lines to evaluate the anticancer potential of the EA fraction. Impact on cell cycle distribution was determined using flow cytometry. The fraction was investigated for interfering microtubules assembly in both in vitro and cellular assay. Further studies were conducted to determine the fraction induced cell death (apoptosis) using calcein/propidium iodide dual staining, activated caspase-3 and phosphorylation of Bcl-2 protein at Ser70. DNA fragmentation assay was performed to confirm the apoptosis.

Results: EA fraction exhibited potent inhibition of cancer cell growth and resulted in 50% growth inhibition (GI50) of 0.27 μg/mL in A673 cell line. Sarcoma (MG-63, Saos-2) and ovarian (SK-OV-3 and OVCAR-3) cancer cell lines also showed superior anticancer activity GI50 of 1.0 μg/mL. Colon and breast cancer cell lines exhibited moderate GI compare other cancer cell lines and normal human lung fibroblast showed GI50 of 15.6 μg/mL. EA fraction showed potent G2/M phase arrest in A673 cell line and induced apoptosis at 48 h exposure. EA fraction promoted microtubule polymerization in tubulin polymerization assay and increased level of polymerized tubulin in the HeLa cells. Fraction induced the activation of caspase-3 and phosphorylation of Bcl-2 anti-apoptotic protein. Fraction induced DNA fragmentation in HeLa cells as evidence of apoptosis.

Conclusion: Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.

No MeSH data available.


Related in: MedlinePlus