摘要目的 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.

abstractsObjective 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.