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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

DNA fragmentation assay performed with different concentration of ethyl acetate fraction exposed HeLa cells and Isolated DNA samples were analyzed using (2%) agarose gel electrophoresis
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Figure 9: DNA fragmentation assay performed with different concentration of ethyl acetate fraction exposed HeLa cells and Isolated DNA samples were analyzed using (2%) agarose gel electrophoresis

Mentions: From the above experiments, it was prudent that the EA fraction is inducing the apoptosis through tubulin polymerization. In order to confirm the apoptosis, a hallmark assay DNA ladder assay has been performed. DNA fragmentation is the final process of apoptosis, and this explains the DNA fragmentation between the nucleosomes and forms 140–180 base pairs fragments. The EA fraction treated HeLa cell's nucleus has showed laddering pattern compared to untreated control. This fragmentation was significant in the 3 μg/ml concentration treated cells, but untreated control cells did not show any sign of DNA fragmentation [Figure 9]. Hence, this data gives the confirmation of apoptosis induction in HeLa cells that were treated by EA fraction.


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)

DNA fragmentation assay performed with different concentration of ethyl acetate fraction exposed HeLa cells and Isolated DNA samples were analyzed using (2%) agarose gel electrophoresis
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: DNA fragmentation assay performed with different concentration of ethyl acetate fraction exposed HeLa cells and Isolated DNA samples were analyzed using (2%) agarose gel electrophoresis
Mentions: From the above experiments, it was prudent that the EA fraction is inducing the apoptosis through tubulin polymerization. In order to confirm the apoptosis, a hallmark assay DNA ladder assay has been performed. DNA fragmentation is the final process of apoptosis, and this explains the DNA fragmentation between the nucleosomes and forms 140–180 base pairs fragments. The EA fraction treated HeLa cell's nucleus has showed laddering pattern compared to untreated control. This fragmentation was significant in the 3 μg/ml concentration treated cells, but untreated control cells did not show any sign of DNA fragmentation [Figure 9]. Hence, this data gives the confirmation of apoptosis induction in HeLa cells that were treated by EA fraction.

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