LINC02381调控乳腺癌细胞迁移和侵袭能力的机制研究

Mechanism of LINC02381 in regulating migration and invasion of breast cancer cells

目的探讨长链非编码RNA(long non-coding RNA,lncRNA)LINC02381对乳腺癌细胞迁移和侵袭的分子机制。方法选择人乳腺癌MCF-7、MDA-MB-231细胞,将其分为小干扰RNA(small interfering RNA LINC02381,si-LINC02381)组、阴性对照siRNA NC组和let-7i-5p模拟物(let-7i-5p mimic)组、阴性对照mimic NC组,进行转染;采用RT-PCR检测转染后各组细胞的let-7i-5p水平、KIFC1的mRNA水平;Western blot检测蛋白质表达;CCK-8法检测转染后各组细胞的增殖活力;划痕实验检测转染后各组细胞的划痕愈合率;transwell实验检测转染后各组细胞的侵袭数量。结果通过StarBase v3.0预测发现LINC02381与let-7i-5p之间存在潜在的结合位点,且在1085份乳腺侵袭性肿瘤样本中,两者呈负相关;在MCF-7、MDA-MB-231细胞转染24 h后,与siRNA NC组相比,si-LINC02381组细胞的增殖活力、划痕愈合率和侵袭细胞的数量均显著降低,细胞中let-7i-5p水平升高、KIFC1水平降低;同样转染24 h后,与mimic NC组相比,let-7i-5p mimic组细胞的增殖活力、细胞的划痕愈合率和侵袭细胞的数量降低,细胞中KIFC1水平降低。结论LINC02381可能通过let-7i-5p/KIFC1调控乳腺癌细胞的迁移和侵袭。

Objective To investigate the molecular mechanism of long non-coding RNA(lncRNA)LINC02381 on migration and invasion of breast cancer cells.Methods MCF-7 and MDA-MB-231 cells of human breast cancer were selected and divided into si-LINC02381 group,si-NC group and let-7i-5p mimic group,mimic NC group.qRT-PCR was used to detect the let-7i-5p level and KIFC1 mRNA level of cells in each group after transfection.The protein expression was detected by Western blot.The proliferation activity of transfected cells was detected by CCK-8 method.The scratch healing rate of transfected cells was detected by scratch test,and Transwell assay was used to detect the number of invasion cells in each group after transfection.Results There was a potential binding site between LINC02381 and let-7i-5p predicted by Starbase v3.0,and there was a negative correlation between LINC02381 and let-7i-5p in 1085 breast invasive tumor samples.After transfection of MCF-7 and MDA-MB-231 cells for 24 hours,compared with the negative control group,the proliferation activity,scratch healing rate and the number of invasive cells in si-LINC02381 group decreased significantly,and the level of let-7i-5p increased and the level of KIFC1 decreased.Similarly,24 hours after transfection,compared with the negative control group,the proliferation activity,scratch healing rate and the number of invasive cells in let-7i-5p mimic group decreased,and the level of KIFC1 in cells decreased.Conclusion LINC02381 may can regulate migration and invasion of breast cancer cells through let-7i-5p/KIFC1.