RANBP9调控TGF-β诱导结直肠癌Colo320细胞凋亡的机制研究
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河北北方学院附属第一医院

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河北省科技厅重点研发计划项目(2237784D),2020张家口市级科技计划自筹经费项目(2021050D)


RANBP9 targeted the expression of TGF-β1 induced the cell apoptosis in colorectal cancer Colo320 Cell
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The First Affiliated Hospital of Hebei North University

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    摘要:

    目的:探讨Ran蛋白微管组织中心结合蛋白(RANBP9)靶向调控转化生长因子β1(transforminggrowthfactor-β1, TGF-β1)的表达,及其对结直肠癌Colo320细胞凋亡的影响。方法:分析TCGA数据库中中625例结肠癌组织以及20例正常结肠组织中RANBP9(基因)的表达数据,KMPLOT分析RANBP9的表达与结肠癌患者生存期的关系。人类蛋白免疫组化数据库分析TGF-β1在正常结肠组织和结肠癌组织中的表达情况,使用UALCAN数据库分析TGF-β1的表达与结肠癌患者生存期的关系。双荧光素酶实验分析RANBP9和TGF-β1的关系。将pcDNA3.1-GFP-RANBP9和pcDNA3.1-GFP-RANBP9-NC分别转染至实验组和对照组细胞中,转染完成后细胞常规培养,另设立正常组细胞,不经过转染操作,常规培养;MTT法检测各组细胞的生长活力,流式细胞仪检测各组细胞的凋亡率,JC-1染色检测各组细胞的线粒体膜电位,Western blot检测各组细胞中RANBP9和TGF-β1的表达。结果:RANBP9在结肠癌组织中的表达明显降低,与RANBP9表达低的患者相比,RANBP9高表达的结肠癌患者具有较高的生存期曲线,TGF-β1在结肠癌组织的表达明显升高,与TGF-β1表达高的患者相比,TGF-β1低表达的患者具有较高的生存期曲线,差异均具有统计意义(均P<0.05)。在结肠癌中,RANBP9能靶向调控TGF-β1的表达。与正常组细胞相比,实验组细胞的生长活力、线粒体的膜电位、TGF-β1的表达明显下调,细胞的凋亡率和RANBP9的表达明显升高,差异均具有统计意义(均P<0.05)。结论:RANBP9能靶向调控TGF-β1的表达,促使结肠癌细胞Colo320的生长活力和线粒体膜电位下降,诱导其凋亡。

    Abstract:

    objective: To investigate Ran binding protein in microtubule-organization center (RANBP9) targeted the expression of transforming growth factor β1 (TGF-β1) and its effect on colorectal cancer Colo320 cell apoptosis. Methods: The expression data of RANBP9 (gene)? in 625 cases of colon cancer tissues and 20 cases of normal colon tissues in the TCGA database (TCGA) were analyzed. KMPLOT was used to analyze the relationship between the expression of RANBP9 and the survival time of colon cancer patients. The human protein immunohistochemistry database was used to analyze the expression of TGF-β1 in normal colon tissues and colon cancer tissues. The UALCAN database was used to analyze the relationship between the expression of TGF-β1 and the survival of colon cancer patients.The dual luciferase experiment analyzes the relationship between RANBP9 and TGF-β1.The pcDNA3.1-GFP-RANBP9 and pcDNA3.1-GFP-RANBP9-NC were transfected into the cells in the experimental group and the control group,respectively. And the normal group of cells will be established without transfection operation and routinely cultured.MTT method was used to detect the growth viability of each group of cells, flow cytometry was used to detect the apoptotic rate of each group of cells, JC-1 staining was used to detect the mitochondrial membrane potential of each group of cells, and Western blot was used to detect the expression of RANBP9 and TGF-β1 in each group of cells. Results: The expression of RANBP9 in colon cancer tissue was significantly reduced. Compared with patients with low RANBP9 expression, colon cancer patients with high RANBP9 expression had a higher survival curve, and the expression of TGF-β1 in colon cancer tissue was significantly increased. Compared with patients with high TGF-β1 expression, patients with low TGF-β1 expression had a higher survival curve, and the differences were statistically significant (all P<0.05).In colon cancer, RANBP9 can target the expression of TGF-β1. Compared with the normal group, the growth viability of the experimental group, the mitochondrial membrane potential, and the expression of TGF-β1 were significantly down-regulated, and the apoptosis rate and the expression of RANBP9 were significantly increased. The differences were statistically significant (all P<0.05).Conclusion: RANBP9 can target the expression of TGF-β1, promote the growth of colon cancer cell Colo320 and decrease the mitochondrial membrane potential, and induce its apoptosis.

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  • 收稿日期:2024-01-16
  • 最后修改日期:2024-04-26
  • 录用日期:2024-04-30
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