摘要
目的研究LINC00525靶向miR-338-3p对胃癌细胞凋亡及耐药性的影响。方法将紫杉醇耐药胃癌细胞(HGC-27/PTX)分为si-NC组、si-LINC00525组、miR-NC组、miR-338-3p组、si-LINC00525+anti-miR-NC组、si-LINC00525+anti-miR-338-3p组。细胞计数试剂盒8(cell counting kit 8, CCK-8)检测细胞存活率;实时荧光定量PCR(RT-qPCR)检测LINC00525和miR-338-3p表达水平;蛋白质印迹(Western blot)法检测蛋白表达;Annexin V-FITC/PI双染法检测细胞凋亡;Transwell检测细胞迁移和侵袭;双荧光素酶报告实验验证LINC00525和miR-338-3p的靶向关系。结果 HGC-27和HGC-27/PTX细胞中LINC00525表达水平升高,miR-338-3p表达水平降低。敲除LINC00525和过表达miR-338-3p,caspase-3、P21、E-cadherin表达水平升高,MMP-2表达水平降低,细胞存活率降低,细胞凋亡率升高,细胞迁移、侵袭数降低。LINC00525靶向调控miR-338-3p,抑制miR-338-3p能减弱敲除LINC00525对细胞HGC-27/PTX的存活率、迁移、侵袭及凋亡的影响。结论敲除LINC00525通过上调miR-338-3p表达抑制HGC-27/PTX细胞增殖、迁移和侵袭,促进胃癌细胞凋亡,降低胃癌细胞对紫杉醇的耐药性。
Objective To study the effect of modulating the expression of LINC00525 targeting miR-338-3 p on apoptosis and drug resistance of gastric cancer cells.Methods Paclitaxel-resistant gastric cancer cells(HGC-27/PTX)were transfected with si-NC,si-LINC00525,miR-NC,miR-338-3 p,si-LINC00525+anti-miR-NC,or si-LINC00525+anti-miR-338-3 p.Cell counting kit 8(CCK-8)was used to determine the cell survival rate after the treatments.Real-time fluorescence quantitative PCR(RT-qPCR)was performed to detect LINC00525 and miR-338-3 p expression levels,and Western blotting was used to detect the protein expression in the cells.The changes in cell apoptosis and migration and invasion abilities were analyzed using annexin V-FITC/PI double staining and Transwell assays.Dual luciferase reporter assay was performed to verify the targeting relationship between LINC00525 and miR-338-3 p.Results The expression of LINC00525 was increased and that of miR-338-3 p was decreased in both in HGC-27 and HGC-27/PTX cells.Knocking down LINC00525 and overexpression of miR-338-3 p significantly increased the expression of caspase-3,P21,and E-cadherin,lowered the expression of MMP-2,decreased the cell survival rate,increased the cell apoptotic rate,and suppressed cell migration and invasion.Dual luciferase reporter assay showed that LINC00525 was capable of regulating the targeted miR-338-3 p,and inhibition of miR-338-3 p obviously attenuated the effects of LINC00525 knockdown on the survival,migration,invasion,and apoptosis of HGC-27/PTX cells.Conclusion LINC00525 knockdown can inhibit the proliferation,migration and invasion,promote apoptosis and reduce paclitaxel resistance of HGC-27/PTX cells by up-regulating miR-338-3.
作者
孙玉成
刘晓巍
于红雷
SUN Yucheng;LIU Xiaowei;YU Honglei(First Department of General Surgery,Affiliated Hospital of Yanbian University,Yanji,Jilin Province,133000,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2020年第22期2202-2209,共8页
Journal of Third Military Medical University
作者简介
通信作者:孙玉成,E-mail:ybyysyc@163.com。
