| 杨 乐,李传昌,刘小双,黄建平,刘兆龙.miR-155-TP53INP1调节轴在结直肠癌化疗药物敏感性中的作用机制[J].,2022,(15):2818-2821 |
| miR-155-TP53INP1调节轴在结直肠癌化疗药物敏感性中的作用机制 |
| Mechanism of miR-155-TP53INP1 Axis in Chemosensitivity of Colorectal Cancer |
| 投稿时间：2021-12-28 修订日期：2022-01-24 |
| DOI：10.13241/j.cnki.pmb.2022.15.004 |
| 中文关键词: miR-155 TP53INP1 结直肠癌 耐药 |
| 英文关键词: miR-155 TP53INP1 Colorectal cancer Drug resistance |
| 基金项目:海军军医大学医学免疫学国家重点实验室开放课题（NKLMI-2017K11） |
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| 中文摘要: |
| 摘要 目的：初步揭示miR-155通过靶向调节TP53INP1表达水平影响结直肠癌细胞对5-FU化疗敏感性。方法：将人结肠直肠癌细胞系HCT116进行培养，提取细胞总RNA后，采用miR-155逆转录特异性引物构建反转录体系进行PCR扩增，通过qRT-PCR检测miR-155在5-FU耐药细胞HCT116/FU及敏感细胞株HCT116中的表达情况；取对数生长期细胞，分别转染miR-155mimics、miR-155抑制剂、miR-155阴性对照后，采用CCK-8法检测miR-155对细胞5-FU药物敏感性的影响，双荧光素酶报告基因系统验证miR-155与TP53INP1的靶基因关系，Western blot检测miR-155对 TP53INP1表达的影响。结果：miR-155在HCT116 /Fu细胞中的表达量是HCT116细胞的7.25倍；在相同5-FU浓度时，HCT116+阴性对照的细胞生长抑制率均高于HCT116+mimics、半数抑制浓度显著低于HCT116+mimics，差异均具有统计学意义(P＜0.05)；TP53INP1是miR-155的靶基因，能显著降低野生型TP53INP1 3'-UTR的荧光素酶活性；转染miR-155 mimics后，TP53INP1的相对表达量显著下降，转染miR-155抑制剂后，TP53INP1的相对表达量显著升高，差异均具有统计学意义(P＜0.05)。结论：miR-155水平升高使HCT116细胞对5-FU的敏感性降低，miR-155可能通过靶向调节TP53INP1的表达水平，从而影响结直肠癌细胞对5-FU的敏感性。 |
| 英文摘要: |
| ABSTRACT Objective: To preliminarily reveal the mechanism of miR-155 affects the sensitivity of colorectal cancer cells to 5-FU chemotherapy by targeting the expression level of TP53INP1. Methods: The human colorectal cancer cell line HCT116 was cultured, and the total RNA was extracted, then the reverse transcription system was constructed with miR-155 reverse transcription specific primer for PCR amplification, the expression of miR-155 in HCT116/Fu and HCT116 was detected by qRT PCR; Logarithmic growth cells were transfected with miR-155 mimics, miR-155 inhibitor and miR-155 negative control respectively, the effect of miR-155 on 5-FU drug sensitivity was detected by CCK-8 method, and the dual-luciferase reporter assay was used to verify the relationship between miR-155 and TP53INP1, the effect of miR-155 on the expression of TP53INP1 was detected by Western blot. Results: The expression of miR-155 in HCT116/Fu cells was 7.25 times higher than that in HCT116 cells; At the same 5-FU Concentration, the rate of cell growth inhibition of HCT116+negative control was higher than that of HCT116+mimics, and the half inhibitory concentration was significantly lower than that of HCT116+mimics(P<0.05); TP53INP1 was the target gene of miR-155, which could significantly reduce the luciferase activity of wild-type TP53INP1 3'-UTR; After transfection of miR-155 mimics, the relative expression of TP53INP1 decreased significantly, and after transfection of miR-155 inhibitor, the relative expression of TP53INP1 increased significantly (all P<0.05). Conclusion: The increased level of miR-155 decreased the sensitivity of HCT116 cells to 5-FU, miR-155 may affect the sensitivity of colorectal cancer cells to 5-FU by targeted regulating the expression level of TP53INP1. |
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