中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
5期
1057-1061
,共5页
喻超%孙诚谊%陈美源%肖杰%黎志鹏%江建新
喻超%孫誠誼%陳美源%肖傑%黎誌鵬%江建新
유초%손성의%진미원%초걸%려지붕%강건신
胰腺癌%微小RNA-138-5p%迁移%侵袭%波形蛋白
胰腺癌%微小RNA-138-5p%遷移%侵襲%波形蛋白
이선암%미소RNA-138-5p%천이%침습%파형단백
Pancreatic cancer%MicroRNA%Migration%Invasion%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.
目的 探討微小RNA(miR)-138-5p在胰腺癌組織及細胞中的錶達及其對胰腺癌細胞遷移和侵襲能力的影響及作用機製.方法 以原髮胰腺癌組織、胰腺癌細胞為研究對象,實時熒光定量聚閤酶鏈反應(FQ-PCR)檢測miR-138-5p在胰腺癌組織及細胞中的錶達;miR-138-5p慢病毒過錶達載體(lv-miR-138-m)及抑製載體(lv-miR-138-i)分彆感染胰腺癌PANC-1細胞,細胞劃痕實驗、Transwell細胞侵襲實驗檢測miR-138-5p對PANC-1細胞株遷移、侵襲能力的影響;雙熒光素酶報告基因分型繫統驗證miR-138-5p與波形蛋白(VIM)3'耑非編碼區域(3' UTR)的互補結閤,以證實VIM是miR-138-5p下遊作用靶點.結果 miR-138-5p在8株胰腺癌細胞株中的錶達分彆比正常胰腺細胞低1~ 10倍,在胰腺癌組織中的錶達比癌徬低(52.99±10.62)倍;lv-miR-138-m能明顯抑製胰腺癌PANC-1細胞的遷移、侵襲能力,與陰性對照組比較,分彆降低(1.27 ±0.03)、(4.96±1.16)倍;lv-miR-138-i可明顯增彊其遷移、侵襲能力,分彆提高(3.23±0.71)、(3.40±0.66)倍.miR-138-5p與VIM 3'UTR區域可互補結閤,抑製VIM錶達能減少lv-miR-138-i引起的促進細胞侵襲效應,錶明miR-138-5p至少能部分通過調控VIM而髮揮抑製胰腺癌細胞侵襲的作用.結論 miR-138-5p在胰腺癌細胞的遷移與侵襲性中起重要作用,通過直接靶嚮調控VIM的錶達而抑製胰腺癌細胞侵襲性,這可能是其髮揮抑製胰腺癌細胞侵襲性的機製之一.
목적 탐토미소RNA(miR)-138-5p재이선암조직급세포중적표체급기대이선암세포천이화침습능력적영향급작용궤제.방법 이원발이선암조직、이선암세포위연구대상,실시형광정량취합매련반응(FQ-PCR)검측miR-138-5p재이선암조직급세포중적표체;miR-138-5p만병독과표체재체(lv-miR-138-m)급억제재체(lv-miR-138-i)분별감염이선암PANC-1세포,세포화흔실험、Transwell세포침습실험검측miR-138-5p대PANC-1세포주천이、침습능력적영향;쌍형광소매보고기인분형계통험증miR-138-5p여파형단백(VIM)3'단비편마구역(3' UTR)적호보결합,이증실VIM시miR-138-5p하유작용파점.결과 miR-138-5p재8주이선암세포주중적표체분별비정상이선세포저1~ 10배,재이선암조직중적표체비암방저(52.99±10.62)배;lv-miR-138-m능명현억제이선암PANC-1세포적천이、침습능력,여음성대조조비교,분별강저(1.27 ±0.03)、(4.96±1.16)배;lv-miR-138-i가명현증강기천이、침습능력,분별제고(3.23±0.71)、(3.40±0.66)배.miR-138-5p여VIM 3'UTR구역가호보결합,억제VIM표체능감소lv-miR-138-i인기적촉진세포침습효응,표명miR-138-5p지소능부분통과조공VIM이발휘억제이선암세포침습적작용.결론 miR-138-5p재이선암세포적천이여침습성중기중요작용,통과직접파향조공VIM적표체이억제이선암세포침습성,저가능시기발휘억제이선암세포침습성적궤제지일.
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.