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 MiR-150-Sp is down-regulated in pancreatic cancer. Over- expression of miR-150-Sp inhibits cell proliferation, blocked the cell cycle, but promotes cell apoptosis in pancreatic cancer cells.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
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)