摘要
目的:探讨microRNA-1284(miR-1284)与胃癌的相关性及分子机制。方法:Real-time PCR检测63例胃癌患者的胃癌组织和匹配周围正常胃组织中miR-1284的表达水平及分析其与临床病理特征之间的关系。通过慢病毒载体的构建及包装生成上调miR-1284慢病毒载体并转染胃癌SGC-7901细胞,上调miR-1284后采用realtime PCR验证细胞miR-1284表达水平,CCK-8法检测细胞活性,流式细胞术检测细胞周期和凋亡,划痕实验检测细胞迁移能力,生物信息学软件预测miR-1284潜在的靶基因,real-time PCR测定靶基因JAG1表达水平,Western blotting检测JAG1、Notch1和NF-κB的表达水平。结果:66.67%胃癌患者胃癌组织miR-1284表达明显低于正常胃组织(P<0.05),miR-1284的表达在不同年龄、性别、TNM分期和淋巴结转移情况患者间的差异无统计学意义(P>0.05),但在不同组织学分级患者间的差异有统计学意义(P<0.05)。与LV-NC-GFP组和control组比较,LV-miR-1284组的miR-1284表达水平和细胞凋亡显著升高,细胞周期阻滞于G0/G1期,细胞活性和细胞迁移能力减弱,JAG1、Notch1和NF-κB的表达水平降低,均有显著差异(P<0.05)。结论:miR-1284可能通过调控其靶基因JAG1而发挥抑制胃癌发生发展的作用。
AIM：To evaluate the correlation between microRNA-1284 （miR-1284） and gastric cancer, and to investigate the underlying mechanism.METHODS： The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer （ GC） tissue samples and 63 non-malignant adjacent tissue samples.The correlation between miR-1284 and the clinicopathological feature of GC was analyzed.Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells.After transfection, the expression of miR-1284 was examined by real-time PCR.The cell activity was evaluated by CCK-8 assay.The cell cycle and apoptosis were determined by flow cytometry.The ability of cell migra-tion was measured by wound-healing assay.The potential target gene of miR-1284 was predicted by online bioinformatic softwares.The expression of JAG1 mRNA was examined by real-time PCR.The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting.RESULTS：Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples （ P〈0.05 ） .The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases （P〉0.05）, but significantly associated with histologic grading （P〈0.05）.Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly in-creased （P〈0.05）, the percentages of apoptotic cells and the cells in G0/G1 phase were significantly increased （P〈0.05）, the cells activity and ability of migration were significantly decreased （P〈0.05）, and the expression of JAG1, Notch1 and NF-κB was significantly decreased （P〈0.05）.CONCLUSION：The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2015年第3期440-446,共7页
Chinese Journal of Pathophysiology
基金
广西科学技术攻关项目(No.桂科攻14124004-1-9)
