角果木内生真菌活性分子对HeLa细胞增殖凋亡的影响

Effects of Bio-active Molecules Isolated from Endophytic Fungi of Ceriops tagal on the Proliferation and Apoptosis of HeLa Cells

为了解红树林植物角果木(Ceriops tagal)内生壳囊孢属真菌Cytospora sp.提取物WP-1[(22E,24R)-5,8-epidioxy-5α,8α-ergosta-6,9(11),22E-trien-3β-ol]抑制人宫颈癌HeLa细胞的效果,笔者采用MTT比色法、克隆形成实验、Hoechst 33258染色和蛋白免疫印迹法来研究WP-1对HeLa细胞增殖和凋亡的影响。结果表明,随着WP-1浓度增加,HeLa细胞的存活率显著降低,而且该提取物能明显抑制HeLa细胞的克隆形成,当其浓度达到10μmol·L−1时,HeLa细胞出现染色质浓缩以及核小体碎裂的凋亡现象。蛋白免疫印迹法结果显示,随着WP-1剂量的增加,与凋亡相关的蛋白Caspase-3及Caspase-9的表达下调,Cleaved Caspase-3,Cleaved Caspase-9的表达明显上调。本实验结果显示,WP-1可以抑制HeLa细胞的增殖,并促进HeLa细胞发生凋亡。

In order to understand the effect of WP-1[(22E,24R)-5,8-epidioxy-5α,8α-ergosta-6,9(11),22Etrien-3β-ol]isolated from Cytospora sp.,an endophytic fungus from the mangrove plant Ceriops tagal on human cervical cancer HeLa cells,the proliferation and apoptosis of HeLa cells treated with WP-1 was determined and analyzed by using MTT colorimetry,clone formation,Hoechst 33258 staining and Western blot.It was found that with the increase of WP-1 concentration the survival rate of the HeLa cells decreased significantly,and that the WP-1 significantly inhibited the colony formation of cells.When treated with WP-1 at a concentration of 10μmol·L−1 the HeLa cells showed chromatin condensation and nucleosome fragmentation,which were characteristic of apoptosis.Western blot analysis showed that with the increase of WP-1 dose,the expressions of apoptosis-related proteins Caspase-3 and Caspase-9 were down-regulated in the HeLa cells,while the expressions of Cleaved Caspase-3 and Cleaved Caspase-9 were significantly upregulated.The results show that WP-1 can inhibit the proliferation of HeLa cells and promote apoptosis of HeLa cells.