茯苓均一多糖的分离纯化及其硫酸化衍生物对人乳腺癌MDA-MB-231细胞迁移的影响
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摘要
SATB1在乳腺癌的侵袭转移中起到决定性作用,抑制SATB1的表达能够有效地控制乳腺癌的转移。该实验从茯苓中分离纯化得到均一多糖并制备硫酸化衍生物,筛选具有抑制SATB1表达的多糖成分。茯苓经醇提去除小分子成分,水提醇沉后得茯苓粗多糖PPS,PPS经DEAE Sepharose Fast Flow阴离子交换树脂分离和Superdex-75系列凝胶纯化后得PPSW-1,并对PPSW-1进行结构分析、硫酸衍生化和抑制人乳腺癌MDA-MB-231细胞迁移作用考察。结果显示从茯苓中得到一种具有抑制人乳腺癌MDA-MB-231细胞迁移作用的均一多糖PPSW-1,相对分子质量为3. 06×104,结构为1,3连接α-D-半乳聚糖为主链,1,6连接α-D-半乳聚糖为支链的多糖,PPSW-1表现出较强的抑制细胞迁移的作用,经硫酸衍生化后产物Sul-W-1活性较PPSW-1并未有明显的增强,但水溶性有明显增加,并且发现茯苓多糖及其硫酸化衍生物能抑制SATB1的表达。该研究从茯苓中成功分离到具有抑制人乳腺癌MDA-MB-231细胞迁移作用的均一多糖,并经过硫酸衍生化得到活性不变溶解性更好的多糖,该发现为茯苓多糖及其衍生物的抗乳腺癌研究提供依据。
SATB1 plays a crucial role in the invasion and metastasis of breast cancer,and inhibition of SATB1 expression can effectively control breast cancer metastasis. In this study,homogeneous polysaccharides were isolated from Poria cocos and their sulfated derivatives were prepared to screen out the polysaccharide compositions with inhibitory effects on SATB1 expression. Smal-molecule components were removed from P. cocos by ethanol extraction,and P. cocos crude polysaccharide PPS was obtained by water extraction and ethanol precipitation. Then PPS was successively separated by DEAE Sepharose fast flow anion-exchange and Superdex-75 gel permeation chromatographic steps to give PPSW-1. The structure of PPSW-1 was identified and its sulfated derivatives were prepared. Then their inhibitory effects on human breast cancer MDA-MB-231 cells were investigated. A kind of polysaccharide,PPSW-1 with inhibitory effect on human breast cancer MDA-MB-231 cells,was obtained from P. cocos,with a relative molecular weight of 3. 06×104,and structure of 1,6-branched 1,3-α-D-galactan. PPSW-1 and its sulfated derivative Sul-W-1 showed good inhibitory effect on cells migration,and the water solubility of Sul-W-1 was better than that of PPSW-1. In addition,it was found that polysaccharide of P. cocos and its sulfated derivative can inhibit expression of SATB1. In this study,a kind of homogeneous polysaccharide with inhibitory effect on human breast cancer MDA-MB-231 cells was isolated from P. cocos,and its sulfated derivative with similar efficacy but better solubility was prepared,laying the foundation for the substance basis study of P. cocos.
SATB1 plays a crucial role in the invasion and metastasis of breast cancer,and inhibition of SATB1 expression can effectively control breast cancer metastasis. In this study,homogeneous polysaccharides were isolated from Poria cocos and their sulfated derivatives were prepared to screen out the polysaccharide compositions with inhibitory effects on SATB1 expression. Smal-molecule components were removed from P. cocos by ethanol extraction,and P. cocos crude polysaccharide PPS was obtained by water extraction and ethanol precipitation. Then PPS was successively separated by DEAE Sepharose fast flow anion-exchange and Superdex-75 gel permeation chromatographic steps to give PPSW-1. The structure of PPSW-1 was identified and its sulfated derivatives were prepared. Then their inhibitory effects on human breast cancer MDA-MB-231 cells were investigated. A kind of polysaccharide,PPSW-1 with inhibitory effect on human breast cancer MDA-MB-231 cells,was obtained from P. cocos,with a relative molecular weight of 3. 06×104,and structure of 1,6-branched 1,3-α-D-galactan. PPSW-1 and its sulfated derivative Sul-W-1 showed good inhibitory effect on cells migration,and the water solubility of Sul-W-1 was better than that of PPSW-1. In addition,it was found that polysaccharide of P. cocos and its sulfated derivative can inhibit expression of SATB1. In this study,a kind of homogeneous polysaccharide with inhibitory effect on human breast cancer MDA-MB-231 cells was isolated from P. cocos,and its sulfated derivative with similar efficacy but better solubility was prepared,laying the foundation for the substance basis study of P. cocos.
引文
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[13] Chaidedgumjorn A,Toyoda H,Woo E R,et al. Effect of(1→3)-and(1→4)-linkages of fully sulfated polysaccharides on their anticoagulant activity[J]. Carbohyd Res,2002,337(10):925.
[14] Joseph J P,Harishankar M K,Pillai A A,et al. Hypoxia induced EMT:a review on the mechanism of tumor progression and metastasis in OSCC[J]. Oral Oncol,2018,80:23.
[15]赵亮,丁彦青.结直肠癌转移机制研究进展[J].浙江大学学报:医学版,2014,43(4):486.
[16]陈亭亭,杨培伟,张树辉.芹菜素抗肿瘤机制的研究进展[J].中国现代应用药学,2019,36(4):507.
[17]刘云霞,徐叶峰,姚勇伟,等.益气补肾方含药血清对人胃癌细胞SGC-7901增殖转移及CD44+EpCAM+干细胞比例的影响[J].中国现代应用药学,2018,35(12):1805.
[18]余彬彬,钱金娥,曹剑波.小檗胺对肝癌SMMC-7721细胞侵袭转移的影响[J].中国现代应用药学,2018,35(12):1771.
[19] Kobierzycki C,Wojnar A,Dziegiel P. Expression of SATB1 protein in the ductal breast carcinoma tissue microarrays-preliminary study[J]. Folia Histochem Cyto,2013,51(4):333.
[20] Han H J,Russo J,Kohwi Y,et al. SATB1 reprogrammes gene expression to promote breast tumour growth and metastasis[J].Nature,2008,452(7184):187.
[21]尹婕,李琦,孙立东,等.天然产物抗癌研究进展[J].中国中药杂志,2019,44(1):19.
[22]焦延娜,韩淑燕.抗癌中药单体对肿瘤细胞自噬的调控[J].中国实验方剂学杂志,2017,23(7):206.
[2] Huang J,Li H,Ren G. Epithelial-mesenchymal transition and drug resistance in breast cancer[J]. Int J Oncol,2015,47(3):840.
[3] Han H J,Russo J,Kohwi Y,et al. SATB1 reprogrammes gene expression to promote breast tumour growth and metastasis[J].Nature,2008,452(7184):187.
[4] Kohwishigematsu T,Poterlowicz K,Ordinario E,et al. Genome organizing function of SATB1 in tumor progression[J]. Semin Cancer Biol,2013,23(2):72.
[5]张敏,高晓红,孙晓萌,等.茯苓的药理作用及研究进展[J].北华大学学报,2008,9(1):63.
[6]金勇.不同茯苓菌丝体的多糖结构和生物活性[D].武汉:武汉大学,2004.
[7] Xu Y,Dong Q,Qiu H,et al. Structural characterization of an arabinogalactan from Platycodon grandiflorum roots and antiangiogenic activity of its sulfated derivative[J]. Biomacromolecules,2010,11(10):2558.
[8]王辉俊,柯樱,叶冠.活性导向分离瓜蒌皮中具有抗血管紧张素转化酶作用的成分[J].中国中药杂志,2017,42(16):3131.
[9] Xu Y X,Dong Q,Qiu H,et al. A homogalacturonan from the radix of Platycodon grandiflorum and the anti-angiogenesis activity of poly-/oligogalacturonic acids derived therefrom[J]. Carbohyd Res,2011,346(13):1930.
[10] Xu Y,Dong Q,Qiu H,et al. Structural characterization of an arabinogalactan from Platycodon grandiflorum roots and antiangiogenic activity of its sulfated derivative[J]. Biomacromolecules,2010,11(10):2558.
[11] Wang H,Shi S,Gu X,et al. Homogalacturonans from preinfused green tea:structural characterization and anticomplementary activity of their sulfated derivatives[J]. J Agr Food Chem,2013,61(46):10971.
[12] Inoue K,Kawamoto K,Nakajima H,et al. Chemical modification and antitumor activity of a D-manno-D-glucan from microellobosporia grisea[J]. Carbohyd Res,1983,115:199.
[13] Chaidedgumjorn A,Toyoda H,Woo E R,et al. Effect of(1→3)-and(1→4)-linkages of fully sulfated polysaccharides on their anticoagulant activity[J]. Carbohyd Res,2002,337(10):925.
[14] Joseph J P,Harishankar M K,Pillai A A,et al. Hypoxia induced EMT:a review on the mechanism of tumor progression and metastasis in OSCC[J]. Oral Oncol,2018,80:23.
[15]赵亮,丁彦青.结直肠癌转移机制研究进展[J].浙江大学学报:医学版,2014,43(4):486.
[16]陈亭亭,杨培伟,张树辉.芹菜素抗肿瘤机制的研究进展[J].中国现代应用药学,2019,36(4):507.
[17]刘云霞,徐叶峰,姚勇伟,等.益气补肾方含药血清对人胃癌细胞SGC-7901增殖转移及CD44+EpCAM+干细胞比例的影响[J].中国现代应用药学,2018,35(12):1805.
[18]余彬彬,钱金娥,曹剑波.小檗胺对肝癌SMMC-7721细胞侵袭转移的影响[J].中国现代应用药学,2018,35(12):1771.
[19] Kobierzycki C,Wojnar A,Dziegiel P. Expression of SATB1 protein in the ductal breast carcinoma tissue microarrays-preliminary study[J]. Folia Histochem Cyto,2013,51(4):333.
[20] Han H J,Russo J,Kohwi Y,et al. SATB1 reprogrammes gene expression to promote breast tumour growth and metastasis[J].Nature,2008,452(7184):187.
[21]尹婕,李琦,孙立东,等.天然产物抗癌研究进展[J].中国中药杂志,2019,44(1):19.
[22]焦延娜,韩淑燕.抗癌中药单体对肿瘤细胞自噬的调控[J].中国实验方剂学杂志,2017,23(7):206.