miR-1275通过靶向〖STBX〗IGF1R〖STBZ〗基因对鼻咽癌细胞增殖和凋亡的影响及作用机制
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湛江市科技计划项目(2020B01033))


The effect of miR1275 on the proliferation and apoptosis of nasopharyngeal carcinoma cells by targeting IGF1R gene
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    摘要:

    【摘要】目的 探讨miR1275靶向胰岛素样生长因子1受体(IGF1R)对鼻咽癌细胞增殖、凋亡的影响及作用机制。方法 通过在线生物信息学软件预测miR1275和IGF1R的靶向关系,采用双荧光素酶报告基因实验进行验证。Western blot检测转染miR1275 模拟物(mimics)或抑制物(inhitior)后的鼻咽癌HONE1细胞IGF1R表达。MTT法、流式细胞术及Western blot检测miR1275/IGF1R分子轴对HONE1细胞增殖、凋亡及蛋白激酶B(AKT)、磷酸化的蛋白激酶B(pAKT)、细胞增殖核抗原( PCNA)和活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase3)表达的影响。结果 HONE1细胞转染miR1275 mimics后,miR1275表达明显升高,细胞增殖活力明显降低,凋亡率明显升高,pAKT和PCNA表达明显降低,cleaved caspase3表达明显升高(P<005)。miR1275可靶向作用于IGF1R并下调其表达。抑制IGF1R表达可明显降低HONE1细胞增殖活力,促进细胞凋亡,下调pAKT和PCNA,上调cleaved caspase3表达(P<005)。抑制miR1275和IGF1R表达可逆转IGF1R对细胞增殖、凋亡及pAKT、PCNA和cleaved caspase3表达的影响。结论 过表达miR1275可抑制鼻咽癌HONE1细胞增殖,促进细胞凋亡,抑制AKT信号途径。IGF1R是miR1275的靶基因,miR1275可靶向IGF1R并调控AKT信号途径影响鼻咽癌细胞增殖和凋亡,miR1275/IGF1R分子轴可能成为鼻咽癌治疗的重要靶点。

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    【Abstract】Objective To investigate the effect and mechanism of miR1275 targeting Insulin like growth factor 1 receptor(IGF1R) on the proliferation and apoptosis of Nasopharyngeal carcinoma cells. Methods The targeting relationship between miR1275 and IGF1R was predicted by online bioinformatics software and verified by double luciferase reporter gene experiment. Western blotting was used to detect the expression of IGF1R in HONE1 cells transfected with miR1275 mimics or miR1275 inhibitor. MTT, flow cytometry and Western blotting were used to detect the effects of miR1275/ IGF1R axis on HONE1 cell proliferation, apoptosis and expression of protein kinase B(tAKT), Phosphorylated protein kinase B(pAKT), Proliferating cell nuclear antigen(PCNA) and cleaved caspase 3. Results The expression of miR1275 was significantly increased after miR1275 mimics were transfected into HONE1 cells, the cell proliferation activity was significantly decreased, the apoptosis rate was significantly increased, the expression of pAKT and PCNA was significantly decreased, and the expression of cleaved caspase 3 was significantly increased (P<005). miR1275 can target IGF1R and down regulate its expression. Inhibition of IGF1R expression can significantly reduce the proliferation of HONE1 cells, promote cell apoptosis, down regulate pAKT and PCNA, up regulate the expression of cleaved cysteinyl aspartate specific proteinase 3(cleaved caspase 3) (P<005), while inhibition of miR1275 and IGF1R expression can reverse the effects of IGF1R on cell proliferation, apoptosis and the expression of pAKT, PCNA and cleaved caspase 3. Conclusion miR1275 can promote the proliferation and inhibit the apoptosis of NPC cells by up regulating the expression of IGF1R.

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  • 在线发布日期: 2021-03-10
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