Annamalai Pazhanimuthu, Thayman Malini, Rajan Sowmiya, Raman Lakshmi Sundaram, Ramasubbu Sankar, Perumal Pachiappan
Department of Biotechnology, Periyar University, Karuppur, Salem, India.
Department of Cell and Molecular Biology, Central Research Facility, Sri Ramachandra University, Porur, Chennai, India.
Pharmacogn Mag. 2015 Apr-Jun;11(42):345-55. doi: 10.4103/0973-1296.153088.
Marine sponges are important sources of bioactive compounds.
This study investigated the anticancer properties of Hyattella cribriformis ethyl acetate (EA) fraction in various cancer and normal cell lines.
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.
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.
Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.
海洋海绵是生物活性化合物的重要来源。
本研究调查了筛板海亚特海绵乙酸乙酯（EA）提取物在多种癌细胞系和正常细胞系中的抗癌特性。
在15种细胞系中进行抗癌试验，以评估EA提取物的抗癌潜力。使用流式细胞术确定其对细胞周期分布的影响。在体外和细胞试验中研究该提取物对微管组装的干扰作用。进一步研究使用钙黄绿素/碘化丙啶双重染色、活化的半胱天冬酶-3以及Bcl-2蛋白Ser70位点的磷酸化来确定该提取物诱导的细胞死亡（凋亡）。进行DNA片段化分析以确认凋亡。
EA提取物对癌细胞生长具有强效抑制作用，在A673细胞系中导致50%生长抑制（GI50）为0.27μg/mL。肉瘤（MG-63、Saos-2）和卵巢（SK-OV-3和OVCAR-3）癌细胞系也显示出优异的抗癌活性，GI50为1.0μg/mL。结肠和乳腺癌细胞系与其他癌细胞系相比表现出中等程度的GI，而正常人肺成纤维细胞的GI50为15.6μg/mL。EA提取物在A673细胞系中显示出强效的G2/M期阻滞，并在暴露48小时时诱导凋亡。EA提取物在微管蛋白聚合试验中促进微管聚合，并增加HeLa细胞中聚合微管蛋白的水平。该提取物诱导半胱天冬酶-3的活化以及抗凋亡蛋白Bcl-2的磷酸化。该提取物在HeLa细胞中诱导DNA片段化，作为凋亡的证据。
海洋海绵筛板海亚特海绵EA提取物通过微管蛋白聚合和诱导凋亡表现出强效抗癌活性。
