摘要
目的构建、筛选并鉴定siRNA-ATF3表达载体,探讨转录激活因子3(ATF3)、乳腺丝氨酸蛋白酶抑制因子(Maspin)和基质金属蛋白酶2(MMP2)在转染了siRNA-ATF3表达载体的人胶质母细胞U373MG细胞株中的表达和意义。方法构建siRNA-ATF3表达载体,并转染U373MG细胞株,以荧光定量PCR和Western blot检测筛选出有效的siRNA-ATF3表达载体,以免疫细胞化学法检测转染siRNA-ATF3表达载体的U373MG细胞株中ATF3、Maspin和MMP2的蛋白表达情况。结果U373MG细胞中的ATF3蛋白表达量可被pSilencer siR1-ATF3和pSilencer siR2-ATF3表达载体有效降低。转染siRNA-ATF3表达载体后U373MG细胞生长状态差,增殖速度减慢。ATF3和MMP2在空白组和Control-siRNA组中的阳性表达量高于3个干扰组;与空白组和Control-siRNA组相比,Maspin在ATF3-siRNA组表达最高,在顺铂组和ATF3-siRNA+顺铂组中的表达降低。结论siRNA-ATF3表达载体有效抑制U373MG细胞的生长和增殖,顺铂可协同增强抑制效果,ATF3、Maspin和MMP2可能协同参与了U373MG细胞的生长和增殖,siRNA-ATF3表达载体可作为治疗抑制胶质母细胞瘤生长增殖的有效手段。
Objective To construct,screen and identify the expression vector of sirna-atf3,and investigate the expression and significance of ATF3,Maspin and MMP2 in human glioblast U373MG cell line transfected with siRNA-ATF3 expression vector.Methods The siRNA-AFT3 expression vector was constructed and transfected into U373MG cell line.The effective siRNA-AFT3 ex⁃pression vector was screened by fluorescence quantitative PCR and Western blot.The expressions of ATF3,Maspin and MMP2 in U373MG cell line transfected with siRNA-AFT3 expression vector was detected by immunocytochemistry.Results Psilencer siR1-ATF3 and psilencer siR2-ATF3 expression vectors can effectively reduce the protein expression level of ATF3 gene in U373MG cells.After transfection with siRNA-AFT3 expression vector,U373MG cells grew poorly and the proliferation rate was slowed down.The number of ATF3 and MMP2 positive cells in ATF3 siRNA group,cisplatin group and ATF3 siRNA+cisplatin group were less than those in blank group and control siRNA group;compared with blank group and control siRNA group,maspin expression was the highest in ATF3 siRNA group,and decreased in cisplatin group and ATF3 siRNA+cisplatin group.Conclusion siRNA-ATF3 ex⁃pression vector can effectively inhibit the growth and proliferation of U373MG cells.Cisplatin can synergistically enhance the inhibito⁃ry effect.ATF3,Maspin and MMP2 may be involved in the growth and proliferation of U373MG cells.siRNA-ATF3 expression vector can be used as an effective method to treat the growth and proliferation of glioblastoma.
作者
马斯奇
孙逸杰
朱旭强
李雪元
吴立新
闫东明
MA Siqi;SUN Yijie;ZHU Xuqiang;LI Xueyuan;WU Lixin;YAN Dongming(The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《中国实用神经疾病杂志》
2021年第13期1110-1116,共7页
Chinese Journal of Practical Nervous Diseases
基金
河南省科技攻关计划(国际科技合作)项目资助(编号:172102410015)。
