重组人Hespintor对肝癌细胞增殖、迁移和侵袭的抑制作用  

Inhibitory effects of recombinant human Hespintor on proliferation,migration and invasion of hepatocellular carcinoma cells

作　　者：孙杰[1] 赵梓晗 潘志鹏[2] 潘凌鸿[1] 伦永志[1,2] Sun Jie Zhao Zihan Pan Zhipeng Pan Linghong Lun Yongzhi(Faculty of Laboratory Medical Science, School of Pharmacy and Medical Technology, Putian University, Putian, Fujian 351100, China Faculty of Laboratory Medical Science, College of Medicine, Dalian University, Dalian , Liaoning 116622, China)

机构地区：[1]莆田学院药学与医学技术学院医学检验系,福建莆田351100 [2]大连大学医学院医学检验系,辽宁大连116622

出　　处：《重庆医学》2017年第9期1182-1185,共4页Chongqing medicine

摘　　要：目的完善重组Hespintor蛋白(rHespintor)的纯化方法,提高蛋白提取效率,并探讨其对肝母细胞瘤HepG2细胞增殖及侵袭作用的影响。方法在重组蛋白提取中增加包涵体洗涤过程,更改蛋白纯化缓冲体系,并利用Na-苯甲酰-DL-精氨酸-对硝基酰胺盐酸盐(BAPNA)为作用底物,测定纯化的rHespintor对胰蛋白酶水解抑制活性。以空白组为对照组,通过MTT实验、细胞划痕愈合实验、肿瘤细胞侵袭实验分别检测rHespintor对肝母细胞瘤HepG2细胞生长的影响及作用效果。结果对包涵体蛋白进行尿素梯度洗涤后可有效去除目的蛋白中绝大多数杂蛋白,一步纯化后,目的蛋白rHespintor具有较高胰蛋白酶水解抑制活性,且抑制效果呈剂量依赖关系。rHespintor作用于肝母细胞瘤HepG2细胞后,细胞增殖受到抑制,迁移能力减弱,侵袭的细胞数量显著减少。结论 rHespintor在体外可明显抑制肝母细胞瘤HepG2细胞的增殖及侵袭作用。Objective To perfect the purification method of recombinant fusion protein of Hespintor（rHespintor）for increasing the protein extraction efficiency,and to investigate its effects on the proliferation,migration and invasion of hepatoblastoma cell line HepG2.Methods In the recombinant protein extraction,the inclusion body washing process was added and the protein purification buffer system was changed.BAPNA was used as the substrate.The inhibitory effect tof purified rHespintor on trypsin hydrolysis was detected.The blank group served as the control group.The MTT test,cell scratch wound healing test and tumor cell invasion test were performed to detect the effect of rHespintor on growth of hepatoblastoma HepG2 cells and its effect.ResultsThe urea gradient washing on the inclusion body protein could effectively remove the vast majority of impure proteins from the targeted protein.After one-step purification,the target protein rHespintor exhibited a high inhibition effect of trypsin hydrolysis,and the inhibitory effect was exhibited a dose-dependent manner.After acting on hepatoblastoma HepG2 cells with rHespintor,the cell proliferation ability was inhibited,the migration ability was reduced and the number of invaded cells were significantly decreased.Conclusion rHespintor can significantly inhibit the proliferation,migration and invasion of hepatoblastoma cell line HepG2 cells in vitro.

关 键 词：重组蛋白质类 丝氨酸蛋白酶抑制因子 细胞增殖 细胞运动 细胞侵袭 

分 类 号：R735.7[医药卫生—肿瘤]