摘要
目的了解胰腺癌中异常表达的微小RNA（miRNA）对胰腺癌细胞增殖和凋亡的影响。方法利用miRNA芯片比较胰腺癌组织、胰腺癌细胞株ASPC-1和正常胰腺组织的miRNA表达谱。根据表达谱结果选择4个异常表达的miRNA：miR-21、24、191、221，利用脂质体转染反义核苷酸方法下调这些miRNA的活性。荧光显微镜监测转染效率；荧光定量PCR检测miRNA的表达。MTT法细胞活力分析、流式细胞仪分析下调miRNA后的细胞活力、生长周期和凋亡的变化。结果下调miR．21增加细胞凋亡，降低细胞活性；下调miR-221改变细胞周期，增加G0／G1期细胞，减少S期细胞，但对细胞活性无明显影响；miR-221和miR-21共同下调后，细胞活力则明显下降。其他miR转染对细胞生长和凋亡的影响不明显。结论部分在胰腺癌中异常表达的miRNA对胰腺癌细胞的生长有一定影响，选用一定组合的联合转染，对细胞生长和凋亡的影响比单独转染时加强。
Objective To investigate the effect of abnormally expressed miRNA on proliferation and apoptosis in pancreatic cancer. Methods miRNA microarray was used to determine the miRNA profiles in pancreatic cancer tissue, pancreatic cancer cell line ASPC-1, and normal pancreatic tissues. According to the profiles, miRNA mir-21,24, 191, 221 were selected, then the activities of these miRNA were down-regulated by method of lipofection of antisense oligonucleotides. Transfection efficiency was measured by fluorescence microscope ; fluorescence quantitative PCR was used to observe the expression of miRNA. Cell viability analysis was done by MTT, and flow cytometry was employed for cell viability analysis, cell cycle and apoptosis after miRNAs were down-regulated. Results miR-21 down-regulation alone increased cell apoptosis and decreased cell viability; while miR-221 down-regulation alone changed the cell cycle and increased G0G1 phase cell, but had no effect on cell viability; when both miR-21 and miR-221 were down-regulated, cell viability was significantly decreased, but other miRNA had no significant effects on cell proliferation and apoptosis. Conclusions Some abnormally expressed miRNAs could affect the proliferation of pancreatic cancer cell, co- transfection of certain combination of miRNAs had enhanced effects on the proliferation and apoptosis of pancreatic cancer cell than any single miRNA transfection.
出处
《中华胰腺病杂志》
CAS
2009年第3期170-173,共4页
Chinese Journal of Pancreatology
基金
广东省自然科学基金（7001208）
