蒋淳琪,王 丹,洪善贻.miRNA-375靶向调控Notch1基因对胰腺癌细胞增殖、迁移的作用机制研究[J].肿瘤学杂志,2020,26(6):501-505. |
miRNA-375靶向调控Notch1基因对胰腺癌细胞增殖、迁移的作用机制研究 |
MiRNA-375 Regulates Proliferation and Migration of Pancreatic Cancer Cells through Targeting Notch1 Gene |
投稿时间:2019-09-20 |
DOI:10.11735/j.issn.1671-170X.2020.06.B007 |
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中文关键词: miRNA-375 胰腺肿瘤 Notch1 增殖 靶向 |
英文关键词:miRNA-375 pancreatic cancer Notch1 proliferation targeting |
基金项目:国家自然科学基金(81302932) |
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中文摘要: |
摘 要:[目的] 探讨miRNA-375靶向调控Notch1基因对胰腺癌(PAAD)细胞增殖、迁移的作用机制。[方法] 选取诊断为PAAD并行胰腺外科手术患者60例,取癌组织和癌旁组织行HE染色、免疫组织化学法和免疫荧光检测,采用PCR法检测胰腺癌组织中miRNA-375和Notch1表达量,使用transwall侵袭实验法和MTT试剂盒检测胰腺癌细胞增殖、迁移和侵袭性,采用生物信息学分析和预测miRNA-375调控胰腺癌细胞Notch1的表达。[结果] miRNA-375表达水平在癌组织(1.00±0.07)与癌旁组织(1.65±0.14)中有显著性差异(P<0.05);miRNA-375表达水平与肿瘤大小、TNM分期、淋巴结转移和远处转移有关(P<0.05)。qRT-PCR检测显示PANC-1细胞和BxPC-3细胞中miRNA-375 mimics表达水平显著增加(P<0.05),转染3天后PANC-1细胞和BxPC-3细胞增殖被显著抑制(P<0.05),抑制率分别为35.93%和27.71%;转染miRNA-375使其表达水平上升可显著抑制PANC-1细胞和BxPC-3细胞的迁移能力。miRNA-375高表达使PANC-1细胞水解基质胶穿透到小室下面的细胞减少35.35%,BxPC-3细胞减少33.04%(P<0.05)。miRNA-375可以调控Notch1D 表达。利用双荧光素酶报告基因法检测miRNA-375与Notch1 mRNA的作用关系,转入突变3′UTR质粒的细胞中荧光素酶活性显著降低(t=5.633,P=0.039;t=5.347,P=0.031),miRNA-375直接与Notch1 mRNA的3′UTR相互作用。[结论]胰腺癌细胞中miRNA-375靶向调控Notch1表达。miRNA-375/Notch1通路能够抑制胰腺癌细胞的增殖、成瘤及侵袭转移能力,在胰腺癌细胞中发挥抑癌作用。 |
英文摘要: |
Abstract:[Objective] To investigate the effect of miRNA375 on proliferation and migration of pancreatic cancer cells and its mechanism. [Methods] The expression of miRNA375 and Notch1 in 60 samples of pancreatic cancer and adjacent tissues were detected by RT-PCR. The proliferation,migration and invasion were detected by MTT method and Transwall assay,respectively. The regulatory effect of microRNA375 on Notch1 gens was predicted by bioinformatics analysis and verified by luciferase reporter gene method. [Results] The expression level of miRNA-375 in cancer tissues was lower than that in adjacent tissues(1.00±0.07 vs 1.65±0.14,P<0.05). The expression of miRNA-375 was correlated with tumor size,TNM stage,lymph node metastasis and distant metastasis(P<0.05). qRT-PCR showed that the expression level of miRNA-375 in PANC-1 cells and BxPC-3 cells increased significantly(P<0.05). After 3 days of transfection,the proliferation of PANC-1 cells and BxPC-3 cells were significantly inhibited(P<0.05),and the inhibition rates were 35.93% and 27.71%,respectively. The expression level of miRNA-375 increased with the transfection of miRNA-375,it can significantly inhibit the migration of pancreatic cancer PANC-1 cells and BxPC-3 cells. The high expression of miRNA-375 reduced the penetration of hydrolyzed matrix glue of PANC-1 cells into cells under the chamber by 35.35%,and that of BXPC-3 cells by 33.04%(P<0.05). Using double luciferase reporter gene method to detect the relationship between miRNA-375 and Notch 1 mRNA,the luciferase activity in the mutant 3′UTR plasmid cells decreased significantly(t=5.633,P=0.039;t=5.347,P=0.031),which proved that miRNA-375 directly interacted with Notch 1 mRNA 3′UTR. [Conclusion] The expression of Notch1 is specifically regulated by miRNA375 in pancreatic cancer cells. MicroRNA-375/Notch1 pathway may inhibit the proliferation,tumorigenesis,invasion and metastasis of pancreatic cancer cells,and play an anti-cancer role in pancreatic cancer cells. |
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