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中华临床医师杂志(电子版) ›› 2023, Vol. 17 ›› Issue (08) : 884 -890. doi: 10.3877/cma.j.issn.1674-0785.2023.08.007

基础研究

敲低IMPDH1对结肠癌SW480、HT29细胞生物功能的影响
王飞飞, 王光林, 孟泽松, 李保坤, 曹龙飞, 张娟, 周超熙, 丁源一, 王贵英()   
  1. 050010 石家庄,河北医科大学第四医院外二科
    050010 石家庄,河北医科大学第四医院外二科;050061 石家庄,河北医科大学第二医院
  • 收稿日期:2022-01-03 出版日期:2023-08-15
  • 通信作者: 王贵英
  • 基金资助:
    河北省医学科学研究课题计划(20210054)

Effect of IMPDH1 knockdown on biological behavior of human colon cancer SW480 and HT29 cells

Feifei Wang, Guanglin Wang, Zesong Meng, Baokun Li, Longfei Cao, Juan Zhang, Chaoxi Zhou, Yuanyi Ding, Guiying Wang()   

  1. The Second Department of General Surgery, The Fourth Hospital, Hebei Medical University, Shijiazhuang 050010, China
    The Second Department of General Surgery, The Fourth Hospital, Hebei Medical University, Shijiazhuang 050010, China; The Second Hospital, Hebei Medical University, Shijiazhuang 050051, China
  • Received:2022-01-03 Published:2023-08-15
  • Corresponding author: Guiying Wang
引用本文:

王飞飞, 王光林, 孟泽松, 李保坤, 曹龙飞, 张娟, 周超熙, 丁源一, 王贵英. 敲低IMPDH1对结肠癌SW480、HT29细胞生物功能的影响[J]. 中华临床医师杂志(电子版), 2023, 17(08): 884-890.

Feifei Wang, Guanglin Wang, Zesong Meng, Baokun Li, Longfei Cao, Juan Zhang, Chaoxi Zhou, Yuanyi Ding, Guiying Wang. Effect of IMPDH1 knockdown on biological behavior of human colon cancer SW480 and HT29 cells[J]. Chinese Journal of Clinicians(Electronic Edition), 2023, 17(08): 884-890.

目的

探讨敲低IMPDH1后对人结肠癌SW480、HT29细胞生物功能的影响。

方法

通过qRT-PCR法验证人体正常结肠粘膜细胞FHC及结肠癌细胞SW480、HT29中IMPDH1基因的表达情况;利用siRNA敲低SW480、HT29细胞中IMPDH1的表达,采用qRT-PCR法及Western blot法检测SW480、HT29细胞中IMPDH1的mRNA及蛋白表达水平,验证转染效果;通过MTT、Transwell及划痕愈合实验检测敲低IMPDH1基因表达后,对SW480、HT29的细胞增殖、侵袭及迁移能力的影响。

结果

IMPDH1在FHC、SW480及HT29细胞中均有表达,且IMPDH1在SW480、HT29细胞中的表达高于FHC细胞;siRNA转染SW480、HT29细胞后,IMPDH1在mRNA及蛋白表达水平与对照组相比,差异具有统计学意义(P<0.01);根据MTT的实验结果显示,敲低IMPDH1的表达能抑制SW480、HT29细胞的体外增殖能力(P<0.05);Transwell的实验结果显示,敲低IMPDH1的表达能抑制SW480、HT29细胞体外侵袭能力(P<0.05);划痕愈合实验显示,敲低IMPDH1的表达,能抑制SW480、HT29细胞的体外迁移能力(P<0.05)。

结论

IMPDH1在SW480、HT29细胞中的表达水平高于在FHC细胞中的表达水平;敲低IMPDH1表达后,在体表抑制SW480、HT29的细胞增殖、侵袭及迁移能力。

Objective

To investigate the effect of IMPDH1 knockdown on the biological behavior of human colon cancer SW480 and HT29 cells.

Methods

The expression of IMPDH1 gene in the human normal colon mucosal cell line FHC and the human colon cancer cell lines SW480 and HT29 was detected by qRT-PCR. Small interfering RNAs (siRNAs) was used to knock down the expression of IMPDH1 in SW480 and HT29 cells, and the mRNA and protein expression levels of IMPDH1 in SW480 and HT29 cells were detected by qRT-PCR and Western blot, respectively, to verify the transfection effects. MTT, Transwell, and scratch healing assays were performed to detect the effects of IMPDH1 gene knockdown on the proliferation, invasion, and migration of SW480 and HT29 cells.

Results

IMPDH1 was expressed in FHC, SW480, and HT29 cells, and the expression of IMPDH1 in SW480 and HT29 cells was higher than that in FHC cells. After siRNA transfection of SW480 and HT29 cells, the mRNA and protein expression levels of IMPDH1 were significantly reduced compared with those of the control group (P<0.05). MTT assay showed that IMPDH1 knockdown inhibited the proliferation of SW480 and HT29 cells in vitro (P<0.01). Transwell assay showed that IMPDH1 knockdown inhibited the invasiveness of SW480 and HT29 cells in vitro (P<0.05). The scratch healing assay showed that IMPDH1 knockdown inhibited the migration ability of SW480 and HT29 cells (P<0.05).

Conclusion

IMPDH1 expression in SW480 and HT29 cells is higher than that in FHC cells. IMPDH1 could inhibit the proliferation, invasion, and migration of SW480 and HT29 cells in vitro.

表1 IMPDH1特异性靶向和非靶向对照siRNA序列
表2 内参及IMPDH1引物序列
图1 qRT-PCR检测各细胞中IMPDH1的相对表达水平
图2 qPCR法检测结肠癌细胞中基因IMPDH1 mRNA表达水平
图3 Western blot法检测结肠癌细胞中IMPDH1蛋白表达水平
图4 敲低IMPDH1表达对SW480、HT29细胞增殖活性的影响
图5 敲低IMPDH1基因表达对SW480、HT29细胞侵袭能力的影响
图6 a 敲低IMPDH1基因表达对结肠癌SW480细胞迁移能力的影响
图6 b 敲低IMPDH1基因表达对结肠癌HT29细胞迁移能力的影响
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