中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2013年
7期
460-464
,共5页
孙洋%金香兰%张婷婷%贾从伟%陈杰
孫洋%金香蘭%張婷婷%賈從偉%陳傑
손양%금향란%장정정%가종위%진걸
胰腺肿瘤%微RNAs%肿瘤细胞,培养的
胰腺腫瘤%微RNAs%腫瘤細胞,培養的
이선종류%미RNAs%종류세포,배양적
Pancreatic neoplasms%MicroRNAs%Tumor cells,cultured
目的 探讨miR-150-5p对胰腺癌细胞生长及凋亡的影响.方法 即时定量PCR(qRT-PCR)法检测11例胰腺癌及对应癌旁正常组织和4个胰腺癌细胞系中miR-150-5p的表达,脂质体介导的化学合成miR-150-5p类似物转染PANC-1、MIA PaCa-2、BxPC-3和AsPC-1细胞系,并采用qRT-PCR法在PANC-1、MIAPaCa-2细胞系中检测转染效率；采用CCK-8检测miR-150-5p对胰腺癌细胞生长的影响；在PANC-1和MIA PaCa-2中分别使用PI/FITC双染法和PI单染,流式细胞术检测miR-150-5p对细胞周期和凋亡的影响.结果 miR-150-5p在11例胰腺癌组织中的表达低于癌旁正常胰腺组织,在4种细胞系中的表达低于正常胰腺组织(均P ＜0.05).转染miR-150-5p mimics可有效提高4种胰腺癌细胞中miR-150-5p RNA量(P＜0.01).转染72 h后miR-150-5p可显著抑制AsPC-1、BxPC-3、MIA PaCa-2和PANC-1细胞生长,与正常对照组相比,抑制率分别为50.7％、48.6％、30.8％和42.3％(P＜0.01).与对照组比miR-150-5p能够促进胰腺癌细胞凋亡(P＜0.01).MIAPaCa-2中与对照组比miR-150-5p能够显著升高G1期细胞百分比(P＜0.01),MIA PaCa-2和PANC-1中与对照组比miR-150-5p能够显著降低S期细胞百分比(均P＜0.01).结论 miR-150-5p在胰腺癌中表达下调,过表达miR-150-5p可以抑制胰腺癌细胞生长,阻断细胞周期,促进细胞凋亡.
目的 探討miR-150-5p對胰腺癌細胞生長及凋亡的影響.方法 即時定量PCR(qRT-PCR)法檢測11例胰腺癌及對應癌徬正常組織和4箇胰腺癌細胞繫中miR-150-5p的錶達,脂質體介導的化學閤成miR-150-5p類似物轉染PANC-1、MIA PaCa-2、BxPC-3和AsPC-1細胞繫,併採用qRT-PCR法在PANC-1、MIAPaCa-2細胞繫中檢測轉染效率；採用CCK-8檢測miR-150-5p對胰腺癌細胞生長的影響；在PANC-1和MIA PaCa-2中分彆使用PI/FITC雙染法和PI單染,流式細胞術檢測miR-150-5p對細胞週期和凋亡的影響.結果 miR-150-5p在11例胰腺癌組織中的錶達低于癌徬正常胰腺組織,在4種細胞繫中的錶達低于正常胰腺組織(均P ＜0.05).轉染miR-150-5p mimics可有效提高4種胰腺癌細胞中miR-150-5p RNA量(P＜0.01).轉染72 h後miR-150-5p可顯著抑製AsPC-1、BxPC-3、MIA PaCa-2和PANC-1細胞生長,與正常對照組相比,抑製率分彆為50.7％、48.6％、30.8％和42.3％(P＜0.01).與對照組比miR-150-5p能夠促進胰腺癌細胞凋亡(P＜0.01).MIAPaCa-2中與對照組比miR-150-5p能夠顯著升高G1期細胞百分比(P＜0.01),MIA PaCa-2和PANC-1中與對照組比miR-150-5p能夠顯著降低S期細胞百分比(均P＜0.01).結論 miR-150-5p在胰腺癌中錶達下調,過錶達miR-150-5p可以抑製胰腺癌細胞生長,阻斷細胞週期,促進細胞凋亡.
목적 탐토miR-150-5p대이선암세포생장급조망적영향.방법 즉시정량PCR(qRT-PCR)법검측11례이선암급대응암방정상조직화4개이선암세포계중miR-150-5p적표체,지질체개도적화학합성miR-150-5p유사물전염PANC-1、MIA PaCa-2、BxPC-3화AsPC-1세포계,병채용qRT-PCR법재PANC-1、MIAPaCa-2세포계중검측전염효솔；채용CCK-8검측miR-150-5p대이선암세포생장적영향；재PANC-1화MIA PaCa-2중분별사용PI/FITC쌍염법화PI단염,류식세포술검측miR-150-5p대세포주기화조망적영향.결과 miR-150-5p재11례이선암조직중적표체저우암방정상이선조직,재4충세포계중적표체저우정상이선조직(균P ＜0.05).전염miR-150-5p mimics가유효제고4충이선암세포중miR-150-5p RNA량(P＜0.01).전염72 h후miR-150-5p가현저억제AsPC-1、BxPC-3、MIA PaCa-2화PANC-1세포생장,여정상대조조상비,억제솔분별위50.7％、48.6％、30.8％화42.3％(P＜0.01).여대조조비miR-150-5p능구촉진이선암세포조망(P＜0.01).MIAPaCa-2중여대조조비miR-150-5p능구현저승고G1기세포백분비(P＜0.01),MIA PaCa-2화PANC-1중여대조조비miR-150-5p능구현저강저S기세포백분비(균P＜0.01).결론 miR-150-5p재이선암중표체하조,과표체miR-150-5p가이억제이선암세포생장,조단세포주기,촉진세포조망.
Objective To investigate the role of miR-150-5p in cell proliferation and apoptosis in human pancreatic cancer cell lines.Methods The expression of miR-150-5p in pancreatic cancer was detected by real time qPCR analysis in 11 pairs of pancreatic cancer tissue and matched adjacent normal tissue samples and in 4 pancreatic cancer cell lines.PANC-1,MIA PaCa-2,BxPC-3 and AsPC-1 cells were transfected with chemically synthesized MiR-150-5p mimics,and CCK-8 assays was then performed to assess cellular functions.To fully understand the mechanisms by which miR-150-5p exerted its function,cell cycle analysis was performed on MIA PaCa-2 and PANC-1 cells 48 hours after transfection,by incubating with propidium iodide (PI)and subsequently analyzed by fluorescence-activated cell sorting (FACS).Apoptosis assay was performed on MIA PaCa-2 and PANC-1 cell lines 24 hours after transfection using the Annexin V-FITC Apoptosis Detection Kit I (BD Biosciences) and analyzed by FACS.Results The expression of miR-150-5p was consistently lower in the pancreatic cancer tissues than in normal tissues,and the miR-150-5p was also down-regulated in pancreatic cancer cell lines (P ＜ 0.05).MiR-150-5p mimics transfection siguificantly raised the expression level of miR-150-5p mRNA in PANC-1 and MIA PaCa-2 (P ＜0.01).The CCK-8 proliferation assay showed that cell growth was reduced in 4 pancreatic cancer cell lines (AsPC-1,BxPC-3,MIA PaCa-2,PANC-1) of miR-150-5p transfected cells compared with NC-transfected cells.The inhibition rates were 50.7％,48.6％,30.8％ and 42.3％,respectively (P ＜ 0.01).The apoptotic rate was increased in cells transfected with miR-150-5p mimics (P ＜0.01).The cell cycle analysis in MIA PaCa-2 indicated that miR-150-5p treatment induced cell cycle arrest in G1 phase with a significant increase in the percentage of cells in G1 phase (P ＜ 0.01),and a reduction of the S-phase cell population in MIA PaCa-2 and PANC-1 (P＜0.01).Conclusions MiR-150-5p is down-regulated in pancreatic cancer.Overexpression of miR-150-5p inhibits cell proliferation,blocked the cell cycle,but promotes cell apoptosis in pancreatic cancer cells.
