联合敲减eIF4E和YB1基因对人宫颈癌细胞增殖、侵袭和迁移能力的影响  

Regulatory effects of co-silenced eIF4E and YB1 on the proliferation,invasion and migration of human cervical cancer cells

作　　者：王晓露 成莹 沐超 胡新荣 WANG Xiaolu;CHENG Ying;MU Chao(School of Basic Medical Science,Guangdong Medical University,Guangdong,Dongguan 523000,China)

机构地区：[1]广东医科大学基础医学院,广东省东莞市523000

出　　处：《河北医药》2023年第18期2725-2730,共6页Hebei Medical Journal

基　　金：国家自然科学基金资助项目(编号:81572566);广东医科大学大学生创新训练项目(编号:202010571030)。

摘　　要：目的 探究在宫颈癌HeLa和C33a细胞中联合敲减YB1和eIF4E基因对细胞增殖、侵袭和迁移行为的影响。方法 将细胞分组并转染(HeLa细胞系:mock空白对照组、NC组(sh-YB1组、Si-eIF4E组、共敲减shYB1+Si-eIF4E组;C33a细胞系:mock空白对照组、NC组、shYB1组、Si-eIF4E组、shYB1+Si-eIF4E组),24 h后RT-qPCR检测转染效率,48 h后western blot检测蛋白表达情况,CCK-8法和平板细胞克隆形成实验检测细胞增殖能力,Transwell法测细胞侵袭迁移能力。结果 敲减YB1和eIF4E基因后mRNA和蛋白表达量明显下降,差异有统计学意义(P<0.05)。细胞增殖实验结果表明,sh-YB1组、Si-eIF4E组及共敲减YB1+eIF4E组与mock组及NC组之间细胞吸光度差异有统计学意义(P<0.05)。敲减YB1和(或)eIF4E基因后,细胞吸光度下降明显(P<0.05),且sh-YB1+Si-eIF4E组吸光度较sh-YB1或Si-eIF4E均减小,差异有统计学意义(P<0.05)。平板细胞克隆形成实验表明,sh-YB1组、Si-eIF4E及共敲减YB1+eIF4E组克隆形成数量明显低于mock组及NC组(P<0.05)。HeLa细胞系、C33a细胞系中与mock组相比,sh-YB1组及Si-eIF4E组和共敲减YB1+eIF4E组平板细胞克隆形成率显著降低(P<0.05)。Transwell实验结果表明,sh-YB1组、Si-eIF4E组和共敲减YB1+eIF4E组相比于mock组及NC组之间穿过小室的细胞明显减少(P<0.05),且共敲减YB1+eIF4E组、sh-YB1组/Si-eIF4E组、mock组、NC组穿过小室的细胞数目依次增加,差异均有统计学意义(P<0.01)。结论 联合敲减YB1和eIF4E基因对宫颈癌细胞的增殖、侵袭和迁移能力有抑制作用。Objective To investigate the regulatory effects of co-silence of Y-box-binding protein-1(YB1)and eukaryotic translation initiation factor 4E(eIF4E)on cell proliferation,invasion,and migration of the cervical cancer cell line HeLa and C33a.Methods HeLa and C33a cells were treated with blank control,or transfected with NC,sh-YB1,si-eIF4E or sh-YB1+si-eIF4E.The transfection efficiency was detected by reverse transcription quantitative PCR(RT-qPCR)at 24h.Protein expressions were detected by Western blot at 48 h.The cell counting kit-8(CCK-8)assays and colony formation assay were performed to detect cell proliferation.Transwell assay was performed to detect cell invasion and migration.Results Co-transfection of sh-YB1 and si-eIF4E significantly downregulated mRNA and protein levels of YB1 and eIF4E(P<0.05).The results of cell proliferation experiments showed that there was a significant difference in the optical density in cells transfected with sh-YB1,si-eIF4E,sh-YB1+si-eIF4E,and mock and those with blank control(NC)(P<0.05).Knockdown of YB1 and/or eIF4E significantly reduced the optical density,which was significantly lower in co-transfected cells than those transfected with sh-YB1 or si-eIF4E(P<0.05).Colony formation assay revealed that the number of colonies in cells transfected with sh-YB1,si-eIF4E,and sh-YB1+si-eIF4E was significantly less than that of cells transfected with mock or NC(P<0.05).In HeLa and C33a cells,the cloning efficiency was significantly lower in cells transfected with sh-YB1,si-eIF4E,and sh-YB1+si-eIF4E than that of cells transfected with mock(P<0.05).Transwell assay revealed that the number of penetrating cells was significantly less in cells transfected with sh-YB1,si-eIF4E,and sh-YB1+si-eIF4E than that of cells transfected with mock or NC(P<0.05).It gradually increased in cells transfected with sh-YB1,si-eIF4E,sh-YB1+si-eIF4E,and mock and NC,with a significant difference(P<0.01).Conclusion Co-silence of YB1 and eIF4E can inhibit the proliferation,invasion,and migration of cervical cancer c

关 键 词：EIF4E YB1 增殖 侵袭 迁移 

分 类 号：R711.74[医药卫生—妇产科学]