摘要
目的:探讨siRNA介导的eIF3b基因沉默对乳腺癌细胞MCF-7侵袭和迁移能力的影响。方法:设计eIF3b基因的小分子干扰RNA(siRNA)片段,脂质体介导转染入乳腺癌细胞株MCF-7,转染分3个组:I组(空白对照组)、II组(转染非特异性对照组)和III组(转染eIF3b-siRNA组)。应用qRT-PCR和Western blotting免疫印迹法检测转染前后eIF3b基因mRNA和蛋白表达水平;运用Matrigel胶模型细胞迁移和侵袭实验检测MCF-7细胞迁移和侵袭能力的变化。结果:与I组(1.09±0.19)、II组(1.05±0.17)相比,III组eIF3b蛋白表达水平(0.45±0.06)明显下降,差异具有统计学意义(P<0.05),I组和II组eIF3b蛋白表达水平比较,差异无统计学意义(P>0.05)。III组eIF3b mRNA相对表达水平为0.54±0.08,明显低于I组(1.12±0.16)和II组(1.08±0.18),差异有统计学意义(P<0.05),I组和II组eIF3b mRNA表达比较,差异无统计学意义(P>0.05)。细胞迁移实验中,III组细胞明显少于I组和II组(P<0.05);细胞侵袭实验中,III组穿过人工基底膜的细胞明显少于I组和II组(P<0.05)。结论:下调eIF3b表达可明显抑制乳腺癌细胞侵袭和迁移。
Objective:To investigate the influence of RNA interference induced silencing of eIF3b gene on migration and invasion potential in breast cancer cells MCF-7.Methods:Human breast cancer cells MCF-7 were divided into control group(I group),scramble group(II group)and si eIF3b group(III group).siRNA sequence targeting eIF3b gene was designed and synthe sized.The siRNA was transfected into breast cancer cells MCF-7 by lipofectamineTM 2000 mediation.The alteration of eIF3b mRNA and protein expression was detected by qRT-PCR and Western blotting.Invasion potential was evaluated by Transwell invasion assay.Cell migrating ability was detected by Transwell migration assay.Results:eIF3b protein expression of III group(0.45±0.06)was lower than that of I group(1.09±0.19)and II group(1.05±0.17)(P<0.05).There was no difference between I group and II group(P>0.05).eIF3b mRNA expression of III group(0.54±0.08)was lower than that of I group(1.12±0.16)and II group(1.08±0.18)(P<0.05).There was no difference between I group and II group(P>0.05).In cell migration assay,the number of cells in III group was less than that in I group and II group(P<0.05).In invasion potential test,the number of cells in III group was less than that in I group and II group(P<0.05).Conclusion:The siRNA targeting eIF3b gene can restrain the migration and invasion of breast cancer cell.
作者
李延辉
方茅
谢敏如
夏明汗
LI Yanhui;FANG Mao;XIE Minru;XIA Minghan(Department of Pathology,Shijie Hospital,Guangdong Dongguan 523290,China;Pathology Department,Guangzhou Medical University,Guangdong Guangzhou 511483,China;Department of Pathology,the First Hospital of Jinan University,Guangdong Guangzhou 510000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2021年第2期201-205,共5页
Journal of Modern Oncology
基金
2019年东莞市社会科技发展(重点)项目(编号:201950715011207)。
