沉默Linc00152对胰腺癌细胞MIA PaCa-2侵袭及迁移的影响

Effect of Silencing Linc00152 on Invasion and Migration of Pancreatic Cancer Cell Line MIA PaCa-2

目的:探讨长链非编码RNA Linc00152对人胰腺癌MIA PaCa-2细胞侵袭及迁移能力的影响。方法:采用逆转录实时荧光定量聚合酶链式反应(RT-qPCR)检测Linc00152在胰腺癌组织(以配对的癌旁组织作为对照)及胰腺癌细胞株(MIA PaCa-2、Bx PC-3及Capan2)中的表达水平,以正常胰腺细胞HPDE作为对照;将Linc00152表达最高的MIA PaCa-2细胞随机分为阴性对照组(转染阴性对照序列si-NC,NC组)及沉默组(转染siRNA抑制序列沉默Linc00152表达),采用Transwell实验检测2组MIA PaCa-2细胞的侵袭及迁移细胞数,采用细胞划痕实验2组MIA PaCa-2细胞的相对划痕愈合率,采用Western blot检测2组MIA PaCa-2细胞的vimentin及N-cadherin蛋白的表达水平。结果:以相应对照比较,胰腺癌组织和细胞株MIA PaCa-2、Bx PC-3及Capan2中Linc00152表达水平升高(P <0. 05或P <0. 01),MIA PaCa-2表达最高;Transwell实验结果显示,MIA PaCa-2的Linc00152沉默组细胞侵袭、迁移细胞数低于NC组(P <0. 01),细胞划痕实验结果显示细胞的相对划痕愈合率低于NC组(P <0. 01),且细胞vimentin及N-cadherin蛋白表达水平均明显低于NC组(P <0. 01)。结论:沉默Linc00152可抑制胰腺癌细胞株MIA PaCa-2侵袭和迁移能力,机制可能与vimentin及N-cadherin蛋白的表达下调有关。

Objective:To investigate the effect of long-chain noncoding RNA Linc00152 on the invasion and migration of human pancreatic cancer MIA PaCa-2 cells.Methods:Reverse transcription real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect Linc00152 expression in pancreatic cancer tissues,its adjacent tissues,pancreatic cancer cell lines(MIA PaCa-2,BxPC-3 and Capan2),and normal pancreatic cell line HPDE.MIA PaCa-2 cells were transfected with siRNA against negative control sequence(NC)or siRNA against Linc00152 expression.Transwell assay was used to detect cell invasion and migration,while scratch assay was used to detect cell healing rate.Western blot was used to detect the protein expression levels of vimentin and N-cadherin.Results:The expression level of Linc00152 was higher in pancreatic cancer tissues than those in adjacent tissues(P<0.05).Similarly,Linc00152 levels were elevated in cell lines MIA PaCa-2,BxPC-3 and Capan2 compared to HPDE cells(P<0.01 or P<0.05).Linc00152 level was the highest in MIA PaCa-2 cells.Transwell assay showed that silencing Linc00152 in MIA PaCa-2 cells impaired cell invasion and migration when compared to NC(P<0.01).Scratch assay exhibited that silencing Linc00152 inhibited cell healing rates relative to NC(P<0.01).Moreover,silencing Linc00152 downregulated the expression levels of vimentin and N-cadherin protein compared to NC(P<0.01).Conclusion:Silencing Linc00152 inhibites the cell invasion and migration of pancreatic cancer cell line MIA PaCa-2.The mechanism may be related to the downregulation of vimentin and N-cadherin.