砷所致恶性转化细胞的外泌体微小RNA-191促肝细胞增殖作用  

Exosomal microRNA-191 derived from arsenite-transformed cells promotes the proliferation of human fiver cells

作　　者：陈超[1] 罗菲[1] 刘欣璐 刘起展[1] 

机构地区：[1]南京医科大学公共卫生学院卫生毒理学系,南京211166

出　　处：《中华地方病学杂志》2017年第2期113-117,共5页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81273114、81430077）;江苏省高等教育优势学科建设项目（2014）

摘　　要：目的探讨亚砷酸钠（NaAsO2）所致恶性转化人肝细胞L-02分泌的外泌体微小RNA（miR）-191对正常人肝细胞L-02增殖的影响。方法分别使用2μmoL／LNaAsO2恶性转化L-02细胞的培养液、提取的外泌体，处理正常的野生型L-02细胞（受体L-02细胞）；利用转染试剂Lipofectamine^TM 2000将anti-miR-191和对照anti-miR-NC分别转染至恶性转化L02（T-L-02）细胞，同时设置未处理组。采用实时荧光定量PCR检测miR-191表达：CCK-8法检测细胞增殖。结果与同代对照L-02细胞培养液（CM）处理组相比，NaAsO2恶性转化细胞培养液（T-CM）处理组显著促进受体L-02细胞增殖[（207±24）％比（105±21）％，t=5．462，P〈0．01]和miR-191表达[（206±25）％比（105±20）％，t=4．116，P〈0．05]。来源于CM的不同浓度外泌体未引起受体L-02细胞增殖和miR-191表达明显变化（F=2．213、2．213，P均〉0．05）；来源于T-CM的不同浓度外泌体引起受体L-02细胞增殖和miR-191表达明显增高，且存在一定剂量．效应关系（F=10．910、4．553，P〈0．01或〈0．05）。20、50mg／L T-CM来源的外泌体处理的受体L-02细胞相对增殖率高于同浓度CM来源的外泌体处理组[（160±32）％比（102±8）％，（203±7）％比（111±5）％，t=2．999、18．750，P〈0．05或〈0．01]，10、20、50mg／L T-CM来源的外泌体处理的受体L-02细胞miR-191表达高于同浓度CM来源的外泌体处理组[（166±13）％比（113±9）％，（211±55）％比（102±8）％，（206±31）％比（105±6）％，t=5．611、3．357、5．509，P〈0．05或〈0．01]。anti—miR-191处理组T—L-02细胞及其分泌的外泌体miR-191表达[（39±10）％、（30±19）％]显著低于未处理组和anti—miR-NC处理组[（100±0）％、（106±17）％，（100±0）％、（104±17）％，P均〈0．01]。未处理组外泌体促进受体L-02细胞相对增殖率升高[（395±31）％比（100±0）％，t=16．Objective To investigate the role of exosomal microRNA（miR）-191 derived from NaAsOE- transformed cells in proliferation of human liver L-02 cells. Methods The normal wild-type L-02 cells （recipient L-02 cells） were treated with media or exosome derived from 2 μmoL/L NaAsO2-transformed L-02 cells. Anti-miR-191 and anti-miR-NC were transfected into NaAsO2-transformed L-02 （T-L-02） cells by lipofeetamineTM 2000, respectively, while untreated group was set as control. The expression of miR-191 was detected by qRT-PCR. Cell proliferation was evaluated by CCK-8 assay. Results The proliferation [（207 ± 24）% vs （105 ± 21）%, t = 5.462, P 〈 0.01] and the expression of miR-191 [（206 ± 25）% vs （105 ± 20）%, t = 4.116, P 〈 0.05] of recipient L-02 cells were significantly increased in the transformed L-02 cells media （T-CM） treated group compared with in the normal L-02 cells media （CM） group. Several concentrations of exosomes derived from CM did not change the proliferation and miR-191 expression of recipient L-02 cells （F = 2.213, 2.213, all P 〉 0.05）. Several concentrations of exosomes derived from T-CM increased the proliferation and miR-191 expression of recipient L-02 cells in a dose-response manner （F = 10.910, 4.553, P 〈 0.01 or 〈 0.05）. The proliferation [（160 ± 32）% vs （102 ± 8）%, （203 ± 7）% vs （111 ± 5）%, t = 2.999, 18.750, P 〈 0.05 or 〈 0.01] of recipient L-02 cells treated with 20 or 50 mg/L exosomes derived from T-CM was higher than that treated with the same concentration of exosomes derived from CM. The expression of miR-191 [（166 ± 13）% vs （113 ± 9）%, （211± 55）% vs （102 ± 8）%, （206 ± 31）% vs （105 ± 6）%, t = 5.611, 3.357, 5.509, P〈 0,05 or 〈 0.01] of recipient L-02 cells treated with 10, 20 or 50 mg/L exosomes derived from T-CM was higher than that treated with the same concentration of exosomes derived from CM. The miR-191 levels of T-L-02 cells [（39 ± 10）% vs （100 ± 0）% or （106 ± 1

关 键 词：砷化物 外泌体 微小RNA-191 增殖 

分 类 号：R599.1[医药卫生—内科学]