摘要
目的利用制备的贻贝多糖Mytilus polysaccharide-I(MP-I)进行体外抗肿瘤活性验证,并探讨其机制。方法水提醇沉法获得贻贝多糖纯品MP-I。培养多种肿瘤细胞,分别以2×105/mL细胞数接种于96孔板,加入终浓度为20、100、500μg/mL的MP-I多糖溶液(分别为MP-I低、中、高剂量组),以及0.9%氯化钠溶液(对照组)、5μg/mL的5-氟尿嘧啶溶液(5-Fu组),采用MTT比色法检测MP-I的体外抗肿瘤活性。对数生长期的MKN-45肿瘤细胞,以5×105/mL细胞数接种于6孔板,Western印迹法检测不同剂量MP-I处理后MKN-45肿瘤细胞P65蛋白表达水平。MKN-45肿瘤细胞接种BALB/c裸鼠,分别腹腔注射0.9%氯化钠溶液(对照组)、5-Fu 50mg·kg-1·d-1(5-Fu组)、MP-I 20和100mg·kg-1·d-1(MP-I低、高剂量组),观察MP-I体内抑瘤作用。结果 MP-I中、高剂量组对MKN-45、MCF-7、HO-8910、K562、SMMC-7721肿瘤细胞的抑制率均显著高于MP-I低剂量组(P值分别<0.01、0.05),MP-I高剂量组对MCF-7、HO-8910、K562肿瘤细胞的抑制率均显著高于MP-I中剂量组(P值分别<0.01、0.05)。5-Fu组和MP-I低、高剂量组的裸鼠体重均显著低于对照组(P值分别<0.01、0.05),MP-I低、高剂量组又显著高于5-Fu组(P值分别<0.01、0.05)。5-Fu组、MP-I低剂量组的瘤重均显著低于对照组(P值均<0.01),MP-I低、高剂量组又显著高于5-Fu组(P值均<0.01)。5-Fu组和MP-I低、高剂量组间抑瘤率的差异均无统计学意义(P值均>0.05)。对照组在培养24和48h的MKN-45肿瘤细胞P65蛋白相对表达量显著高于MP-I低、中、高剂量组(P值均<0.01),后3组间的差异也有统计学意义(P值均<0.01)。对照组脾淋巴细胞活力的光密度值为0.42±0.03,显著低于MP-I低剂量组的0.48±0.04(P<0.05)和MP-I高剂量组的0.59±0.04(P<0.01),MP-I高剂量组与MP-I低剂量组间的差异也有统计学意义(P<0.05)。结论贻贝多糖MP-I对MKN-45、MCF-7、HO-8910、K562、SMMC-7721肿瘤细胞有体外抑制作用,并且在体内能够抑制
Objective To investigate anti-tumor activity and mechanism of polysaccharide isolated from Mytilus coruscus.Methods The Mytilus polysaccharide-I（MP-I）was extracted from Mytilus coruscus.Tumor cells were cultured at 2×105per milliliter in 96 well plates and treated with 500,100 and 20μg/mL MP-I（highdose group,middle-dose group and low-dose group）,0.9% sodium chloride solution（control group）and 5μg/mL5-fluorouracil（5-Fu group）,respectively.The in vitro anti-tumor activity of MP-I was assessed by using methylthiazolyldiphenyl-tetrazolium bromide（MTT）method.Haman gastric cell line（MKN）-45 cells were cultured at 5×105per milliliter in 6well plates and treated with different concentrations of MP-I.The P65 protein level in MKN-45 cells was detected by western blotting.MKN-45 cells were xenografted in Balb/c nude mice.The tumorbearing mice were divided into control group（0.9%sodium chloride solution）,5-Fu group（20mg·kg-1·d-1）and two drug test groups（20mg·kg-1·d-1 MP-I group and 100mg·kg-1·d-1 MP-I group）.Body mass and tumor mass were measured to assess in vivo anti-tumor activity of MP-I.Results The results of MTT showed that the inhibition rates of MKN-45,MCF-7,HO-8910,K562 and SMMC-7721 tumor cells in vitro were significantly higher in MP-I middle and high dose groups than those in MP-I low dose group（P0.01 or P0.05）.The inhibition rates of MCF-7,HO-8910 and K562cells in MP-I high dose group were significantly higher than that of MP-I middle dose group（P0.01 or P0.05）.The body mass of BALB/c nude mice in 5-Fu group,MP-I high and low dose groupswere significantly lower than that in control group（P0.01 or P0.05）.The body mass of BALB/c nude mice in MP-I low and high dose groups were significantly higher than that in 5-Fu group（P0.01 or P0.05）.The tumor mass of 5-Fu group and MP-I low dose group were significantly lower than that of control group（all P0.01）.The tumor mass of MP-I low and high dose groups were significantly higher than that of 5-Fu group（b
出处
《上海医学》
CAS
CSCD
北大核心
2015年第1期57-61,共5页
Shanghai Medical Journal
基金
上海市科学技术委员会长三角联合攻关项目资助(10495810300)
关键词
贻贝
多糖
抗肿瘤作用
Mussels
Polysaccharides
Antitumor action
