Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, India.
Daru. 2013 Dec 19;21(1):72. doi: 10.1186/2008-2231-21-72.
Natural compounds can be alternative sources for finding new lead anti-cancer molecules. Marine algae have been a traditional source for bioactive compounds. Enteromorpha intestinalis and Rhizoclonium riparium are two well distributed saline/brackish water algae from Sundarbans. There's no previous report of these two for their anti-proliferative activities.
Cytotoxicity of the algal methanolic extracts (AMEs) on HeLa cells were assayed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) reduction assay. Morphological examinations were done by Haematoxylin, Hoechst 33258 and Acridine orange staining. DNA fragmentation was checked. Gene expressions of Cysteine aspartate protease (Caspase) 3, Tumor protein (TP) 53, Bcl-2 associated protein X (Bax) were studied by Reverse transcription- polymerase chain reaction (RT-PCR) keeping Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as internal control. Protein expressions were studied for Caspase 3, phospho-p53, Bax, Microtubule associated proteins-1/ light chain B (MAP1/LC3B) by western blot.
The AMEs were found to be cytotoxic with Inhibitory concentration 50 (IC50) values 309.048 ± 3.083 μg/ml and 506.081 ± 3.714 μg/ml for E. intestinalis and R. riparium extracts respectively. Treated cells became round with blebbings with condensed nuclei. Acidic lysosomal vacuoles formation occurred in treated cells. Expression of apoptotic genes in both mRNA and protein level was lowered. Expression of LC3B-II suggested occurrence of autophagy in treated cells.
These two algae can be potent candidates for isolating new lead anticancer molecules. So they need further characterization at both molecular and structural levels.
天然化合物可以成为寻找新的抗癌先导分子的替代来源。海洋藻类一直是生物活性化合物的传统来源。肠浒苔和矶根藻是两种分布广泛的盐水/半咸水藻类,来自孙德尔本斯。目前还没有关于这两种藻类具有抗增殖活性的报道。
通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)还原法测定藻类甲醇提取物(AME)对 HeLa 细胞的细胞毒性。通过苏木精、Hoechst 33258 和吖啶橙染色进行形态学检查。检查 DNA 片段化。通过逆转录-聚合酶链反应(RT-PCR)研究半胱天冬氨酸蛋白酶(Caspase)3、肿瘤蛋白(TP)53、Bcl-2 相关蛋白 X(Bax)的基因表达,以甘油醛 3-磷酸脱氢酶(GAPDH)为内参。通过蛋白质印迹法研究 Caspase 3、磷酸化 p53、Bax、微管相关蛋白 1/轻链 B(MAP1/LC3B)的蛋白表达。
AME 被发现具有细胞毒性,肠浒苔和矶根藻提取物的抑制浓度 50(IC50)值分别为 309.048 ± 3.083μg/ml 和 506.081 ± 3.714μg/ml。处理后的细胞变圆,核浓缩有皱缩。处理后的细胞中形成酸性溶酶体空泡。在 mRNA 和蛋白水平上,凋亡基因的表达均降低。处理细胞中 LC3B-II 的表达表明自噬的发生。
这两种藻类可能是分离新的抗癌先导分子的有力候选物。因此,需要在分子和结构水平上对它们进行进一步的表征。