补肾方药干预过表达、沉默sFRP1的UMR106细胞增殖情况及β-catenin的影响  

The effects of bushen jianpi huoxue recipe on cell activity and β-catenin expression in SFRP1 overexpression and silencing UMR106 cells

作　　者：柴生颋 谢平金 史桐雨[3] 万雷 刘海全 林勇 CHAI Shengting;XIE Pingjin;SHI Tongyu;WAN Lei;Liu Haiquan;Lin Yong(The Affiliated Orthopedics and Trauma Hospital,Guangzhou University of Chinese Medicine,Guangzhou 510240;The Laboratory of Orthopaedics and Traumatology of Chinese Medicine,Lingnan Medical Research Center of Guangzhou University of Chinese Medicine,Guangzhou 510405;Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第三附属医院骨科,广东广州510240 [2]广州中医药大学岭南医学研究中心中医骨伤科学实验室,广东广州510405 [3]广州中医药大学,广东广州510405

出　　处：《中国骨质疏松杂志》2018年第8期984-988,共5页Chinese Journal of Osteoporosis

基　　金：广东省自然科学基金(2015A030313367;2014A030310127;S2013040016828);国家自然科学基金(81302991;81373653;81674004;81673786);广东省科技计划项目(2016A020216024;2013B021800058);广州中医药大学青年英才项目(QNYC20120119);2014广东省"优青计划"项目(yq2014041);2016年度广州中医药大学中医骨伤重点学科开放基金(YB16)

摘　　要：目的针对分泌型卷曲相关蛋白1(s FRP1)基因构建沉默及过表达重组腺病毒载体并结合补肾方药含药血清,观察其对大鼠骨肉瘤细胞(UMR106细胞)增殖和β-连环蛋白(β-catenin)表达的影响。方法构建s FRP1沉默及过表达重组腺病毒载体,制备补肾方药含药血清,将培养好的UMR106细胞分为空白血清+空病毒组、含药血清+空病毒组、空白血清+SFRP1沉默组、含药血清+SFRP1沉默组、空白血清+SFRP1过表达组、含药血清+SFRP1过表达组6组,根据组别不同进行干预,并观察细胞增殖及检测β-catenin蛋白表达。结果与空白血清+空病毒组比较,含药血清+空病毒组、空白血清+SFRP1沉默组、含药血清+SFRP1沉默组的细胞增殖、β-catenin蛋白表达均显著升高。空白血清+SFRP1过表达组、含药血清+SFRP1过表达组的细胞增殖活性、β-catenin蛋白表达降低。其中含药血清+SFRP1沉默组、空白血清+SFRP1过表达组变化最明显。结论沉默s FRP1、补肾方药含药血清可提高UMR106细胞增殖以及β-catenin表达,尤以两者同时干预时的作用最强,而过表达s FRP1则降低UMR106细胞增殖活性,补肾方药含药血清某种程度上可抑制s FRP1的过表达。Objective To observe the effect of sFRP1 gene on the proliferation and the expression of beta-catenin protein in UMR106 cells by constructing sFRP1 silencing and overexpression recombinant adenovirus vector and culturing in serum containing Bushen recipe. Methods We constructed sFRP1 silencing and overexpression recombinant adenovirus vector, prepared prescription containing serum,and cultured UMR106 cells were assigned into six groups including： blank serum ＋ empty virus group,prescription containing serum ＋ empty virus group,blank serum ＋ SFRP1 silencing group,prescription containing serum＋ SFRP1 silencing group,blank serum ＋ SFRP1 overexpression group and prescription containing serum ＋ SFRP1 overexpression group. According to different groups of intervention,we observed the expression of cell proliferation and beta-catenin protein expression. Results compared with blank serum ＋ empty virus group,the cell proliferation and the expression of beta-catenin protein in prescription containing serum ＋ empty virus group,blank serum ＋ SFRP1 silencing group and prescription containing serum ＋ SFRP1 silencing group increased significantly. The cell proliferation activity and the expression of beta-catenin protein in the blank serum ＋ SFRP1 overexpression group and the prescription containing serum ＋ SFRP1 overexpression group decreased.The changes in the prescription containing serum ＋ SFRP1 silencing group and blank serum ＋ SFRP1 overexpression group were the most obvious. Conclusion the silencing of sFRP1 and serum containing kidney tonifying herbs could increase beta-catenin expression and proliferation of UMR106 cells,the strongest effect was observed with both intervention,and over expression of sFRP1 decreased UMR106 cell proliferation,prescription containing serum can inhibit the overexpression of sFRP1 to some extent.

关 键 词：分泌型卷曲相关蛋白1 腺病毒 补肾方药 大鼠骨肉瘤细胞 含药血清 

分 类 号：R285[医药卫生—中药学]