青岛海域浒苔蛋白提取及其抗肿瘤活性的初步研究

A PRELIMINARY STUDY ON EXTRACTION OF PROTION FROM ENTERMORPHA IN QINGDAO SEA AREA AND ITS ANTINEOPLASTIC ACTIVITY

目的提取浒苔蛋白并研究其对肿瘤细胞生长的抑制作用。方法用20mol/L的Tris-HCl(pH8.0)缓冲液悬浮新鲜浒苔,悬浮液反复冻融结合超声波细胞破碎法提取其中的浒苔蛋白。采用四甲基偶氮唑盐比色法(MTT法)研究浒苔蛋白对乳癌细胞EMT6生长的抑制作用,观察肿瘤细胞形态学的变化。建立乳癌细胞EMT6荷瘤小鼠模型,研究浒台蛋白对荷瘤小鼠瘤径和瘤体质量的影响。结果浒苔蛋白粗提率为0.54%,主要是相对分子质量约为25 000的蛋白。随着浒苔蛋白浓度的增加,肿瘤细胞存活率逐渐降低,抑制率逐渐增加;光镜观察结果显示,浒台蛋白作用48h后,肿瘤细胞出现明显的形态学改变。瘤体内注射浒苔蛋白可显著缩小瘤径和瘤体质量,抑瘤率达25%。结论浒苔蛋白在体内、体外均能够抑制肿瘤细胞的增殖,并呈明显的量效关系。

Objective To extract Entermorpha protein （EP） and study its antineoplastic activity. Methods Fresh Entermorpha was suspended in Tris-HCI buffer （20 mmol/L, pH 8.0）, and crude protein was then extracted from the cell lysate by multi-time freeze-thaw followed by ultrasonic treatment. The inhibition effect of EP on breast cancer cells EMT6 was studied by MTT method. A EMT6-bearing mouse model was created, and the effect of EP on the size and weight of the tumor of EMT6-bearing mice was observed. Results The extraction rate of EP was 0.54 % ,the main relative molecular mass being 25,000. Along with the increase of the concentration of EP, the survival rate of the tumor cells decreased and the inhibition rate increased gradually. Light microscopic observation showed that after 48-hour action of the protein, the morphology of the tumor cells changed obviously. Intratumoral injection with EP could notably reduce the diameter and weight of the tumor, the tumor-inhibition rate reaching 25%. Conclusion EP can inhibit the tumor cell proliferation both in vivo and in vitro, which shows a dose-effect relationship.