title: Ethyl acetate extract from marine sponge Hyattella cribriformis exhibit potent anticancer activity by promoting tubulin polymerization as evidenced mitotic arrest and induction of apoptosis.
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.
  
  
  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.
  
  
  CONCLUSIONS
  Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.
authors.affiliation: Department of Biotechnology, Periyar University, Karuppur, Salem, India.
authors.forename: Pazhanimuthu
authors.initials: P
authors.lastname: Annamalai
authors.affiliation: Department of Cell and Molecular Biology, Central Research Facility, Sri Ramachandra University, Porur, Chennai, India.
authors.forename: Malini
authors.initials: M
authors.lastname: Thayman
authors.affiliation: Department of Biotechnology, Periyar University, Karuppur, Salem, India.
authors.forename: Sowmiya
authors.initials: S
authors.lastname: Rajan
authors.affiliation: Department of Cell and Molecular Biology, Central Research Facility, Sri Ramachandra University, Porur, Chennai, India.
authors.forename: Lakshmi Sundaram
authors.initials: LS
authors.lastname: Raman
authors.affiliation: Department of Discovery Biology, AURA Biotechnologies Private Limited, Uthandi, Chennai, Tamil Nadu, India.
authors.forename: Sankar
authors.initials: S
authors.lastname: Ramasubbu
authors.affiliation: Department of Biotechnology, Periyar University, Karuppur, Salem, India.
authors.forename: Pachiappan
authors.initials: P
authors.lastname: Perumal
keywords: Apoptosis and DNA fragmentation; G2/M arrest; Hyattella cribriformis; ethyl extract fraction; microtubule
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