miR-150—5p抑制胰腺癌细胞生长  

MiR-150-5p inhibits the proliferation and promoted apoptosis of pancreatic cancer cells

作　　者：孙洋[1] 金香兰[1] 张婷婷[1] 贾从伟[1] 陈杰[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院病理科,100730

出　　处：《中华病理学杂志》2013年第7期460-464,共5页Chinese Journal of Pathology

基　　金：国家自然科学基金（30270599,30471970,30973470,81172334）;国家科技支撑计划（十一五）项目（2006BA102A14）;卫生部卫生行业科研专项项目（200802011）;教育部高等学校博士学科点专项科研基金（20060023013）

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On．011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、MIAPaCa-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MIAPaCa-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-150-5pmimics可有效提高4种胰腺癌细胞中miR-150-5pRNA量（P〈0．01）。转染72h后miR-150-5p可显著抑制AsPC-1、BxPC-3、MIAPaCa-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），MIAPaCa-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。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-Sp 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 fines. PANC-1, MIA PaCa-2, BxPC-3 and AsPC-1 cells were transfected with chemically synthesized MiR-150-Sp mimics, and CCK-8 assays was then performed to assess cellular functions. To fully understand the mechanisms by which miR-150-Sp 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 transfeetion using the Annexin V-FITC Apoptosis Detection Kit I （BD Bioseiences） 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-Sp mimics transfection significantly raised the expression level of miR-150-Sp 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-Sp treatment induced cell cycle arrest in G1 phase with a significant increase in thepercentage of cells in Gl phase （P 〈0. 01 ）, and PANC-1 （P 〈0. 01）. Conclusions and a reduction of the S-phase cell population in MIA PaCa-2

关 键 词：胰腺肿瘤 微RNAS 肿瘤细胞 培养的 

分 类 号：R735.9[医药卫生—肿瘤]