| 张志友,刘文生,李 鹰.MiR-125b靶向A20/NF-κB信号通路促进胰腺癌细胞增殖机制的研究[J].肿瘤学杂志,2021,27(2):110-115. |
| MiR-125b靶向A20/NF-κB信号通路促进胰腺癌细胞增殖机制的研究 |
| MiR-125b Promotes Proliferation of Pancreatic Cancer Cells by Targeting A20/NF-κB Signaling Pathway |
| 投稿时间：2020-02-21 |
| DOI：10.11735/j.issn.1671-170X.2021.02.B005 |
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| 中文关键词: 胰腺肿瘤 miRNA-125b A20/NF-κB 增殖 |
| 英文关键词:pancreatic cancer miRNA-125b A20/NF-κB proliferation |
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| 中文摘要: |
| 摘 要：［目的］ 研究miR-125b在胰腺癌组织及细胞系中的表达，探讨其对胰腺癌细胞增殖的影响及作用机制。［方法］ QRT-PCR检测miR-125b在60例胰腺癌患者癌组织和癌旁组织中，及3株胰腺癌细胞系和1株正常胰腺导管上皮细胞中的表达；miR-125b敲减慢病毒质粒及阴性对照空载体质粒vector感染SUIT-2细胞，经1.0μg/ml嘌呤霉素筛选稳定敲减miR-125b或vector的SUIT-2细胞，注射到BALB/c裸鼠中，观察抑制miR-125b对胰腺癌细胞体内增殖能力的影响；利用Targetscan 7.2软件及荧光素酶报告基因试验，分析miR-125b对A20基因的靶向作用；SUIT-2细胞转染miR-125b inhibit和scramble后，体外实验检测miR-125b对细胞增殖、肿瘤坏死因子α诱导的蛋白3（tumor necrosis factor α-induced protein 3，TNFAIP3，又称A20）和NF-κB信号通路关键蛋白IKKα/β、IκBα、p65的表达影响。［结果］ MiR-125b在胰腺癌的组织中的表达显著高于癌旁组织 （0.99±0.21 vs 0.68±0.17，P<0.05）；miR-125b在SW1990、SUIT-2、BxPC3胰腺癌细胞系中的表达显著高于在正常胰腺导管上皮细胞HPDE6-C7中的表达（4.81±0.13，8.63±0.27，5.64±0.21 vs 1.00±0.05，P均<0.05）；SUIT-2miR-125b的裸鼠成瘤体积显著低于SUIT-2vector细胞（P<0.05）。TargetScan软件预测和荧光素酶报告基因实验验证A20为miR-125b的靶基因；转染miR-125b inhibit后，A20基因表达显著增加（6.98±0.18 vs 1.00±0.06，P＜0.05），细胞增殖能力、NF-κB信号通路关键蛋白IKKα/β、IκBα、p65的表达显著下降（P均<0.05）。［结论］ MiR-125b可通过靶向调控A20/NF-κB信号通路促进胰腺癌细胞的增殖。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the expression of miR-125b in pancreatic cancer tissues and cell lines，and to explore its effect on the proliferation of pancreatic cancer cells and related mechanism. ［Methods］ The expression of miR-125b in 60 specimens of pancreatic cancer and adjacent tissues，3 pancreatic cancer cell lines and normal pancreatic ductal epithelial cells was detected by qRT-PCR. Human pancreatic carcinoma SUIT-2 cells were transfected with miR-125b-knockdown lentiviral plasmid or with negative control vector，and screened by 1.0 μg/ml puromycin. The SUIT-2 cells with stably knocked down miR-125b or blank vector were injected into BALB/c nude mice，and the size of implanted tumors was measured in two groups. The targeting effect of miR-125b on A20 gene was analyzed by Targetscan 7.2 software and luciferase reporter gene assay. The tumor necrosis factor α-induced protein 3(TNFAIP3 or A20) and NF-κB signaling pathway related proteins IKKα/β，IκBα，p65 were measured with Western blotting. ［Results］ The expression of miR-125b in pancreatic cancer tissue was significantly higher than that in adjacent tissues(0.99±0.21 vs 0.68±0.17，P<0.05).The miR-125b expression in SW1990，SUIT-2，BxPC3 pancreatic cancer cell lines was significantly higher than that in normal pancreatic ductal epithelial cells HPDE6-C7(4.81±0.13，8.63±0.27，5.64±0.21 vs 1.00±0.05，all P<0.05). The tumor size in SUIT-2miR-125b nude mice was significantly lower than that in SUIT-2vector nude mice(P<0.05). TargetScan software and luciferase reporter gene assay indicated that the target gene of miR-125b was A20. After transfection with miR-125b inhibit，A20 gene expression was significantly increased，cell proliferation ability decreased，the expression of NF-κB signaling pathway key proteins IKKα/β，IκBα，p65 decreased significantly(P<0.05). ［Conclusion］ MiR-125b can promote the proliferation of pancreatic cancer cells by targeting A20/NF-κB signaling pathway. |
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