瘤背石磺多糖对人宫颈癌细胞的体外抑制作用  

Inhibitory effect of Onchidium reevesii polysaccharides on Hela cells in vitro

作　　者：贾晶晶 土志涵 练颖康 郁东晨 沈和定[1,2,3] JIA Jingjing;TU Zhihan;LIAN Yingkang;YU Dongchen;SHEN Heding(International Research Center for Marine Biosciences,Ministry of Science and Technology,Shanghai Ocean University,Shanghai 201306,China;Shanghai Universities Key Laboratory of Marine Animal Taxonomy and Evolution,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]上海海洋大学国家海洋生物科学国际联合研究中心,上海201306 [2]上海海洋大学海洋动物系统分类与进化上海高校重点实验室,上海201306 [3]上海海洋大学农业农村部鱼类营养与环境生态研究中心,上海201306

出　　处：《上海海洋大学学报》2023年第2期441-448,共8页Journal of Shanghai Ocean University

基　　金：上海海洋大学水产种质资源研究与支撑平台能力提升(A1-3201-20-300206)

摘　　要：体外试验研究瘤背石磺多糖(Onchidium reevesii polysaccharides, OSP)和硫酸酯化修饰后瘤背石磺多糖(Sulfated Onchidium reevesii polysaccharides, S-OSP)对人宫颈癌Hela细胞生长的影响,并以阳性药物顺铂(0.5、0.75、1.5μg/mL)作为对照。将不同质量浓度(50、75、150μg/mL)的OSP和S-OSP作用于Hela细胞,用CCK-8法分析Hela细胞在12、24、48和72 h的增殖情况,用流式细胞仪分析细胞周期,并应用qRT-PCR检测相关凋亡基因的表达变化。结果表明,OSP和S-OSP均能降低Hela细胞活性并诱导其凋亡,而且S-OSP比OSP更能显著抑制Hela细胞增殖活性,表明硫酸酯化修饰改变了多糖结构进而影响多糖抗肿瘤活性。作用48 h后,3种质量浓度OSP组Hela细胞生长抑制率分别为60%、67%和71%;S-OSP组Hela细胞生长抑制率分别为74%、77%和84%;顺铂组Hela细胞生长抑制率分别为82%、86%和90%。流式细胞术检测显示OSP和S-OSP作用24 h后,细胞早期和晚期凋亡的比率显著,G0/G1期细胞比例增多,S期细胞比例不断降低;实时荧光定量表明促凋亡基因Caspase-3的表达量随药物质量浓度的提高显著增加,抗凋亡基因Bcl-2和Bcl-xL的表达量随药物质量浓度的提高无明显变化,说明OSP和S-OSP可通过调节Caspase信号通路诱导Hela细胞凋亡。In vitro experiments were conducted to study the effects of Onchidium reevesii polysaccharides and sulfated Onchidium reevesii polysaccharides on the growth of Hela human cervical cancer cells,and with positive drug cisplatin(0.5,0.75,1.5μg/mL)as control.Different mass concentrations(50,75,150μg/mL)of OSP and S-OSP were applied to Hela cells,and we used CCK-8 method to analyze the proliferation of Hela cells at 12,24,48 and 72 h,and analyzed the cell cycle with flow cytometry,and used qRT-PCR to detect the expression changes of related apoptosis genes.The results show that both OSP and SOSP can reduce the viability of Hela cells and induce their apoptosis,moreover,S-OSP can more significantly inhibit the proliferation activity of Hela cells than OSP,It shows that the sulfation modification changes the structure of the polysaccharide and affects the anti-tumor activity of the polysaccharide.After 48 hours of treatment,the growth inhibition rates of Hela cells in the three concentrations of OSP group were 60%,67%,and 71%,respectively;the growth inhibition rates of Hela cells in the S-OSP group were 74%,77%,and 84%,respectively;the growth inhibition rates of Hela cells in the cisplatin group were 82%,86%,and 90%,respectively.Flow cytometry showed that after OSP and S-OSP had acted for 24 h,the ratio of early and late apoptosis of cells was significant,and increased proportion of cells in G0/G1 phase,and the proportion of S-phase cells continued to decrease;Real-time fluorescence quantification showed that the expression of pro-apoptotic gene Caspase-3 increased significantly with the increase of drug mass concentration,and the expression of anti-apoptotic genes Bcl-2 and Bcl-xL did not change significantly with the increase of drug mass concentration.It shows that OSP and S-OSP can induce Hela cell apoptosis by regulating the Caspase signaling pathway.

关 键 词：瘤背石磺多糖 硫酸酯化修饰 人宫颈癌细胞 细胞凋亡 实时荧光定量 

分 类 号：R285[医药卫生—中药学]