微小RNA-138-5p靶向调控波形蛋白影响胰腺癌PANC-1细胞迁移和侵袭能力

Effects of microRNA-138-5p on the migration and invasion of pancreatic cancer PANC-1 cells by targeting vimentin

目的 探讨微小RNA（miR）-138-5p在胰腺癌组织及细胞中的表达及其对胰腺癌细胞迁移和侵袭能力的影响及作用机制.方法 以原发胰腺癌组织、胰腺癌细胞为研究对象,实时荧光定量聚合酶链反应（FQ-PCR）检测miR-138-5p在胰腺癌组织及细胞中的表达;miR-138-5p慢病毒过表达载体（lv-miR-138-m）及抑制载体（lv-miR-138-i）分别感染胰腺癌PANC-1细胞,细胞划痕实验、Transwell细胞侵袭实验检测miR-138-5p对PANC-1细胞株迁移、侵袭能力的影响;双荧光素酶报告基因分型系统验证miR-138-5p与波形蛋白（VIM）3＇端非编码区域（3＇ UTR）的互补结合,以证实VIM是miR-138-5p下游作用靶点.结果 miR-138-5p在8株胰腺癌细胞株中的表达分别比正常胰腺细胞低1～ 10倍,在胰腺癌组织中的表达比癌旁低（52.99±10.62）倍;lv-miR-138-m能明显抑制胰腺癌PANC-1细胞的迁移、侵袭能力,与阴性对照组比较,分别降低（1.27 ±0.03）、（4.96±1.16）倍;lv-miR-138-i可明显增强其迁移、侵袭能力,分别提高（3.23±0.71）、（3.40±0.66）倍.miR-138-5p与VIM 3＇UTR区域可互补结合,抑制VIM表达能减少lv-miR-138-i引起的促进细胞侵袭效应,表明miR-138-5p至少能部分通过调控VIM而发挥抑制胰腺癌细胞侵袭的作用.结论 miR-138-5p在胰腺癌细胞的迁移与侵袭性中起重要作用,通过直接靶向调控VIM的表达而抑制胰腺癌细胞侵袭性,这可能是其发挥抑制胰腺癌细胞侵袭性的机制之一.

Objective To explore microRNA-138-5p （miR-138-5p） expression in pancreatic cancer and its effects on pancreatic cancer cell migration and invasion,and its mechanism.Methods Quantitative real-time polymerase chain reaction （FQ-PCR） was used to examine the expression of miR-138-5p in pancreatic cancer cell lines and primary carcinoma tissues from human patients.Lentiviral vector containing miR-138-5p mimics （lv-miR-138-m）,or miR-138-5p inhibitor （lv-miR-138-i） was used to either up-regulate or down-regulate miR-138-5p in PANC-1 cells,respectively.MiR-138-5p impact on PANC-1 cell line migration and invasion abilities by wound-healing and Transwell cell invasion assay.The predicted targeting of miR-138-5p on vimentin was acknowledged by a dual luciferase assay.Results Expression of miR-138-5p in 8 pancreatic cancer cell lines were 1 to 10 times lower than in normal pancreatic cells,expression in pancreatic cancer tissue than in adjacent low 52.99 ± 10.62 times.Lv-miR-138-m significantly inhibited the migration and invasion of PANC-1 cells capability,compared with the negative control group were lower 1.27 ± 0.03,4.96 ± 1.16 times.Lv-miR-138-i can significantly enhance their migration and invasive ability,compared with the negative control group,respectively 3.23 ±0.71,3.40 ±0.66 times.MiR-138-5p and vimentin （VIM） 3＇untranslated region （3＇ UTR） region may be complementary binding,inhibition of expression can reduce the invasiveness effect VIM promote cell lv-miR-138-i induced,suggesting miR-138-5p at least in part through inhibition of pancreatic regulation play VIM cancer cell invasion role.Conclusion MiR-138-5p play an important role in pancreatic cancer cell migration and invasion and it directly targeting regulate the VIM inhibit pancreatic cancer cell invasion,which may be one of the mechanisms which play an inhibition of pancreatic cancer cell invasion.