摘要
目的研究菲律宾蛤仔酶解寡肽(RPO)的制备及体外抗前列腺癌PC-3细胞的活性。方法选用胰蛋白酶对菲律宾蛤仔进行酶解,经截取分子量为3KDa的超滤膜、DEAE SepharoseFF阴离子交换、反相高效液相色谱C18分离制备菲律宾蛤仔酶解寡肽,并应用MTT法、AO/EB染色及Annexin V-FITC/PI双标记流式细胞术检测其体外抗PC-3细胞增殖活性。结果菲律宾蛤仔酶解多肽3KDa以下的组分经阴离子交换,得到的峰1由液相色谱纯化后,获到分子量为607.6 kDa寡肽样品;MTT法结果显示,该寡肽能抑制PC-3细胞增殖,呈剂量和时间依赖;AO/EB染色PC-3出现凋亡的形态学改变;流式细胞仪检测PC-3细胞早期凋亡率、坏死率随寡肽浓度增加而增多,明显高于对照组。结论胰蛋白酶酶解菲律宾蛤仔得到的寡肽能抑制PC-3细胞增殖,诱导凋亡。
Objective To study methods to isolate and purify oligopeptides from Ruditapes philippinarum（RPO） and its antitumor activity against PC-3.Methods Trypsin proteases were applied to hydrolyze the R.philippinarum.Ultrafiltration and anion-exchange chromatography and RP-HPLC were performed to separate and purify the antitumor peptides.MTT assay was used to detect the inhibition effect of the RPO against PC-3 in vitro.AO/EB and Annexin V-FITC/PI was used to detect the apoptosis on PC-3.Results RPO＇s molecular weight is 607.6Da.MTT assay showed that the purified RPO inhibited proliferation of PC-3 cells in both concentration-and time-dependent manners.Flow cytometry studies showed exposing PC-3 cells to RPO for 24h increased the percentage of the early apoptotic cells and cellular necrosis in a dose-dependent manner.In addition,typical morphologic changes were observed in the cells with acridine orange/ethidium bromide staining.Conclusion RPO can inhibit the growth of PC-3 cells and induce apoptosis.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2013年第6期1351-1353,共3页
Lishizhen Medicine and Materia Medica Research
基金
浙江省自然科学基金(No.LY12C20008)
关键词
菲律宾蛤仔
寡肽
PC-3细胞
凋亡
Ruditapes philippinarum
Oligopeptides
PC-3 cell line
Apoptosis
