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中华消化病与影像杂志(电子版) ›› 2025, Vol. 15 ›› Issue (04) : 294 -299. doi: 10.3877/cma.j.issn.2095-2015.2025.04.002

论著

miR-146a靶向PTP1B对结直肠癌细胞增殖、迁移及侵袭的影响
鲁旭娟1, 孔振兴2,()   
  1. 1710003 西安市中心医院检验科
    2710004 西安市人民医院(西安市第四医院)检验科
  • 收稿日期:2025-02-03 出版日期:2025-08-01
  • 通信作者: 孔振兴

Effects of miR-146a targeting PTP1B on proliferation, migration and invasion of colorectal cancer cells

Xujuan Lu1, Zhenxing Kong2,()   

  1. 1Department of Clinical Laboratory, Xi'an Central Hospital, Xi'an 710003, China
    2Department of Clinical Laboratory, Xi'an People's Hospital (Xi'an Fourth Hospital), Xi'an 710004, China
  • Received:2025-02-03 Published:2025-08-01
  • Corresponding author: Zhenxing Kong
引用本文:

鲁旭娟, 孔振兴. miR-146a靶向PTP1B对结直肠癌细胞增殖、迁移及侵袭的影响[J/OL]. 中华消化病与影像杂志(电子版), 2025, 15(04): 294-299.

Xujuan Lu, Zhenxing Kong. Effects of miR-146a targeting PTP1B on proliferation, migration and invasion of colorectal cancer cells[J/OL]. Chinese Journal of Digestion and Medical Imageology(Electronic Edition), 2025, 15(04): 294-299.

目的

探讨微小RNA-146a(miR-146a)靶向蛋白酪氨酸磷酸酶1B(PTP1B)对结直肠癌细胞(HCT-116)增殖、迁移及侵袭的影响。

方法

体外培养结直肠癌细胞系HCT-116细胞和正常结肠上皮细胞株NCM460,将HCT-116细胞随机分为对照组、miR-146a NC组和miR-146a mimics组。实时荧光定量PCR(RT-qPCR)法检测各组HCT-116细胞miR-146a、PTP1B mRNA表达情况;细胞计数试剂盒(CCK-8)法与流式细胞术检测细胞存活率及细胞凋亡情况;划痕实验检测各组HCT-116细胞迁移情况;Transwell小室法检测HCT-116细胞侵袭;蛋白印迹分析法检测各组HCT-116细胞PTP1B、基质金属蛋白酶(MMP)2、MMP-9表达变化;双荧光素酶报告实验验证miR-146a与PTP1B的靶向关系。

结果

与正常结肠上皮细胞NCM460细胞比较,HCT-116细胞miR-146a表达水平显著降低,PTP1B mRNA表达显著升高(P<0.05)。双荧光素酶报告实验证实miR-146a与PTP1B有靶向结合位点。与对照组和miR-146a NC组相比,miR-146a mimics组HCT-116细胞miR-146a表达水平、细胞凋亡率显著升高(P<0.05),细胞存活率、划痕迁移率、侵袭率、PTP1B mRNA和PTP1B蛋白、MMP-2、MMP-9蛋白表达水平显著降低(P<0.05)。

结论

过表达miR-146a可能通过靶向负调控PTP1B表达抑制结直肠癌细胞恶性行为。

Objective

To investigate the effects of microRNA-146a (miR-146a) targeted protein tyrosine phosphatase 1B (PTP1B) on the proliferation, migration and invasion of colorectal cancer cells (HCT-116).

Methods

Human colorectal cancer cell line HCT-116 and normal colon epithelial cell line NCM460 were cultured in vitro, HCT-116 cells were randomly divided into control group, miR-146a NC group and miR-146a mimics group. The expression of miR-146a and PTP1B mRNA in HCT-116 cells was detected by real-time quantitative PCR (RT-qPCR). Cell counting kit (CCK-8) and flow cytometry were used to detect cell survival rate and apoptosis. The migration of HCT-116 cells was detected by scratch test. The invasion of HCT-116 cells was detected by Transwell chamber method. The expression changes of PTP1B, matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blotting. Double luciferase reporter assay was used to verify the targeting relationship between miR-146a and PTP1B.

Results

Compared with those in NCM460 cells, the expression level of miR-146a in HCT-116 cells were significantly lower, and the expression of PTP1B mRNA was significantly higher (P<0.05). Dual luciferase assay confirmed that miR-146a has a targeted binding site with PTP1B. Compared with those in control group and miR-146a NC group, the miR-146a expression level and apoptosis rate of HCT-116 cells in miR-146a mimics group were significantly higher (P<0.05), the cell survival rate, scratch migration rate, invasion rate, expression levels of PTP1B mRNA and protein, MMP-2 and MMP-9 protein were significantly lower (P<0.05).

Conclusion

Overexpression of miR-146a may inhibit malignant behavior of colorectal cancer cells by targeting negative regulation of PTP1B expression.

表1 引物序列
表2 各组细胞miR-146a及PTP1B mRNA表达水平情况( ± sn=6)
图1 miR-146a与PTP1B结合位点预测
表3 各组HCT-116细胞双荧光素酶活性比较( ± sn=6)
表4 miR-146a mimics转染后HCT-116细胞存活率变化( ± sn=6)
图2 各组HCT-116细胞凋亡图片
表5 miR-146a mimics转染后HCT-116细胞的凋亡率( ± sn=6)
图3 各组HCT-116细胞迁移图片(×100)
表6 miR-146a mimics转染HCT-116细胞的迁移率( ± sn=6)
图4 各组HCT-116细胞侵袭图片(结晶紫染色×100)
表7 miR-146a mimics转染HCT-116细胞的侵袭率( ± sn=6)
图5 各组HCT-116细胞PTP1B、MMP-2、MMP-9蛋白表达电泳图注:A对照组;B miR-146a NC组;C miR-146a mimics组
表8 各组细胞PTP1B、MMP-2、MMP-9蛋白表达情况( ± sn=6)
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