医学分子生物学杂志 ›› 2023, Vol. 20 ›› Issue (6): 519-524.doi: 10.3870/j.issn.1672-8009.2023.06.009

• 论著 • 上一篇    下一篇

KIF20A 基因对肝癌 Huh7 细胞放射敏感性的影响及作用机制的研究 #br#

  

  1. 三二〇一医院 1医学检验科,  2肝胆胰腺外科 陕西省汉中市, 723000
  • 出版日期:2023-11-30 发布日期:2023-12-28

Effect of KIF20A Gene on Radiosensitivity of Huh7 Liver Cancer Cells and Its Mechanism #br#

  1. 1Department of Medical Laboratory2Hepatobiliary and Pancreatic Surgery, 3201th Hospital, Hanzhong, Shaanxi, 723000, China
  • Online:2023-11-30 Published:2023-12-28

摘要: 目的 探讨驱动蛋白家族 20A (kinesin family 20A, KIF20A) 基因对肝癌 Huh7 细胞放射敏感性的影响及可能的作用机制方法 qPCR 检测不同剂量 (0、 2、 4、 6、 8 Gy) 照射后 Huh7 KIF20A mRNA表达量, 通过慢病毒感染构建沉默 KIF20A Huh7 细胞株, RT-qPCR 和蛋白质印迹验证 KIF20A 沉默效果,细胞计数试剂盒 8 (cell counting kit 8, CCK-8) 集落形成实验检测细胞增殖, 流式细胞术检测细胞凋亡, Transwell 实验划痕实验检测细胞侵袭和迁移能力, 蛋白质印迹检测 PI3K / AKT 信号通路相关蛋白表达结果 肝癌 Huh7 细胞中 KIF20A mRNA 表达水平随着 X 射线剂量的增加而逐渐上调 (P< 0. 05)。 感染 KIF20A siRNA 能够显著抑制 Huh7 细胞中 KIF20A mRNA 和蛋白表达 (P< 0. 05)。 随着辐射剂量的增加,细胞存活率逐渐下降 (P< 0. 05), 沉默 KIF20A Huh7 细胞存活率明显降低 (P< 0. 05)。 沉默 KIF20A Huh7 细胞集落形成率明显降低 (P< 0. 05), 侵袭和迁移能力明显受到抑制 (P< 0. 05), 凋亡率明显升高(P< 0. 05), 细胞中 PI3K 水平下调 (P< 0. 05), AKT 磷酸化水平减少 (P< 0. 05)。 结论 沉默 KIF20A 能够增强肝癌 Huh7 细胞放射敏感性, 其作用机制可能与 PI3K / AKT 信号通路有关

关键词: KIF20A, PI3K / AKT 信号通路, 肝癌细胞, 放射敏感性

Abstract: Objective To investigate the effect of kinesin family member 20A (KIF20A) gene on the radiosensitivity of hepatocellular carcinoma cells Huh7 and its possible mechanism. Methods RT-qPCR was used to detect the expression levels of KIF20A mRNA in Huh7 after irradiation with different doses (0, 2, 4, 6, 8 Gy). The Huh7 cell line with silenced expression of KIF20A was constructed by lentivirus transduction. The efficiency of RNA silencing on KIF20A was verified byRT-qPCR and Western blotting, cell survival was detected by cell counting kit 8 ( CCK-8) and colony formation assay, apoptosis was detected by flow cytometry, cell invasion and migration were detected by transwell assay and wound-healing assay, and the expression levels of PI3K / AKT signaling pathway-related proteins were detected by Western blotting. Results The mRNA expression level of KIF20A in Huh7 cells was increased with the treatment of increased X-ray dose ( P < 0. 05). KIF20A knockdown significantly inhibited the expression levels of KIF20A mRNA and protein in Huh7 cells (P< 0. 05). With the treatment of increased radiation dose, the cell survival rate decreased gradually (P< 0. 05), the survival rate of Huh7 cells with KIF20A silencing was further decreased (P< 0. 05). The colony formation rate of Huh7 cells with KIF20A silencing was significantly decreased (P< 0. 05), the invasion and migration ability were significantly inhibited (P < 0. 05), the apoptosis rate was significantly increased (P < 0. 05), the level of PI3K in cells was down-regulated (P < 0. 05 ) and the level of AKT phosphorylation was decreased ( P < 0. 05 ). Conclusion Silencing KIF20A can enhance the radiosensitivity of Huh7 liver cancer cells, and its mechanism may be related to the PI3K / AKT signaling pathway.

Key words: KIF20A, PI3K / AKT signaling pathway, liver cancer cells, radiosensitivity

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