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中华消化病与影像杂志(电子版) ›› 2025, Vol. 15 ›› Issue (05) : 436 -443. doi: 10.3877/cma.j.issn.2095-2015.2025.05.004

论著

LncRNA GABPB1-AS1通过靶向hsa-miR-30b-3p促进人结直肠癌细胞增殖侵袭迁移
童笑笑1, 宋美华1, 方政2, 陈峥世1,()   
  1. 1310000 杭州,浙江省立同德医院肛肠外科
    2310000 杭州,浙江省立同德医院消化内科
  • 收稿日期:2025-08-11 出版日期:2025-10-01
  • 通信作者: 陈峥世

LncRNA GABPB1-AS1 promotes the proliferation, invasion and migration of human colorectal cancer cells by targeting hsa-miR-30b-3p

Xiaoxiao Tong1, Meihua Song1, Zheng Fang2, Zhengshi Chen1,()   

  1. 1Department of Anorectal Surgery, Tongde Hospital of Zhejiang Province, Hangzhou 310000, China
    2Department of Gastroenterology, Tongde Hospital of Zhejiang Province, Hangzhou 310000, China
  • Received:2025-08-11 Published:2025-10-01
  • Corresponding author: Zhengshi Chen
  • Supported by:
    Zhejiang Traditional Chinese Medicine Science and Technology Program(2020ZB054)
引用本文:

童笑笑, 宋美华, 方政, 陈峥世. LncRNA GABPB1-AS1通过靶向hsa-miR-30b-3p促进人结直肠癌细胞增殖侵袭迁移[J/OL]. 中华消化病与影像杂志(电子版), 2025, 15(05): 436-443.

Xiaoxiao Tong, Meihua Song, Zheng Fang, Zhengshi Chen. LncRNA GABPB1-AS1 promotes the proliferation, invasion and migration of human colorectal cancer cells by targeting hsa-miR-30b-3p[J/OL]. Chinese Journal of Digestion and Medical Imageology(Electronic Edition), 2025, 15(05): 436-443.

目的

探究LncRNA GABPB1-AS1通过靶向hsa-miR-30b-3p促进人结直肠癌细胞增殖侵袭迁移的分子机制。

方法

体外培养人正常结直肠黏膜细胞系FHC和人结直肠癌细胞系HT-29、SW480、LOVO及WiDr。使用si-NC、si-LncRNA GABPB1-AS1、miR-30b-3p inhibitor对照、miR-30b-3p inhibitor转染HT-29细胞。通过Starbase数据库预测GABPB1-AS1和hsa-miR-30b-3p在结直肠癌组织和癌旁组织中的差异表达和潜在靶向结合位点,GSCA数据库预测GABPB1-AS1表达对结直肠患者生存预后的影响。采用RT-qPCR检测GABPB1-AS1和miR-30b-3p表达水平。采用双荧光素酶实验验证GABPB1-AS1和hsa-miR-30b-3p的靶向关系。采用CCK-8检测细胞活力。采用流式细胞术检测细胞凋亡。采用Transwell实验检测细胞侵袭能力。采用划痕试验检测细胞迁移能力。

结果

生物信息学分析表明,相比癌旁组织,GABPB1-AS1在结直肠癌组织中表达上调,miR-30b-3p表达下调;高表达GABPB1-AS1患者的总生存期和无进展生存期明显缩短。RT-qPCR和双荧光素酶实验结果表明,GABPB1-AS1在结直肠癌组织中表达上调,miR-30b-3p表达下调;GABPB1-AS1靶向负调控miR-30b-3p的表达。敲低GABPB1-AS1的表达能够显著抑制HT-29细胞的活力、侵袭和迁移的能力,促进HT-29细胞的凋亡;在此基础上,抑制miR-30b-3p的表达可部分逆转敲低GABPB1-AS1对HT-29细胞活力、凋亡水平、侵袭和迁移能力的影响。

结论

LncRNA GABPB1-AS1通过靶向负调控hsa-miR-30b-3p的表达促进人结直肠癌细胞增殖、侵袭和迁移。

Objective

To explore the molecular mechanism by which LncRNA GABPB1-AS1 promotes the proliferation, invasion and migration of human colorectal cancer cells by targeting hsa-miR-30b-3p.

Methods

Human normal colorectal mucosal cell line FHC and human colorectal cancer cell lines HT-29, SW480, LOVO and WiDr were cultured in vitro. HT-29 cells were transfected with si-NC, si-LncRNA GABPB1-AS1, miR-30b-3p inhibitor control and miR-30b-3p inhibitor. The differential expression and potential target binding sites of GABPB1-AS1 and hsa-miR-30b-3p in colorectal cancer tissues and adjacent tissues were predicted through the Starbase database, and the impact of GABPB1-AS1 expression on the survival prognosis of colorectal cancer patients was predicted through the GSCA database. The expression levels of GABPB1-AS1 and miR-30b-3p were detected by RT-qPCR. The targeting relationship between GABPB1-AS1 and hsa-miR-30b-3p was verified by dual-luciferase assay. Cell viability was detected by CCK-8. Cell apoptosis was detected by flow cytometry. Cell invasion ability was detected by Transwell assay. Cell migration ability was detected by scratch assay.

Results

Bioinformatics analysis indicated that compared with normal tissues, GABPB1-AS1 was upregulated in colorectal cancer tissues, while miR-30b-3p was downregulated. Patients with high expression of GABPB1-AS1 had significantly shorter overall survival and progression-free survival. The results of RT-qPCR and dual luciferase experiments showed that GABPB1-AS1 was upregulated in colorectal cancer tissues and miR-30b-3p was downregulated. GABPB1-AS1 negatively regulated the expression of miR-30b-3p. Knockdown of GABPB1-AS1 expression could significantly inhibit the viability, invasion and migration abilities of HT-29 cells, and promote the apoptosis of HT-29 cells. On this basis, inhibition of miR-30b-3p expression could partially reverse the effects of knockdown of GABPB1-AS1 on the viability, apoptosis level, invasion and migration ability of HT-29 cells.

Conclusion

The lncRNA GABPB1-AS1 promotes the proliferation, invasion and migration of human colorectal cancer cells by negatively regulating the expression of hsa-miR-30b-3p.

表1 RT-qPCR引物
图1 生物信息学分析注:1A、1B Starbase数据库预测LncRNA GABPB1-AS1和hsa-miR-30b-3p在结直肠癌组织和癌旁组织中的表达水平;1C、1D GSCA数据库预测LncRNA GABPB1-AS1表达对结直肠患者生存预后的影响。COAD结肠腺癌。1A、1B两组间比较采用独立样本t检验,1C、1D采用Log-rank检验
图2 LncRNA GABPB1-AS1和hsa-miR-30b-3p在人结直肠癌细胞中的表达情况注:RT-qPCR检测GABPB1-AS1和miR-30b-3p的表达(n=3);多组间比较采用One-way ANOVA,Tukey's multiple comparisons test分析
图3 LncRNA GABPB1-AS1靶向抑制miR-30b-3p注:3A Starbase数据库预测LncRNA GABPB1-AS1与hsa-miR-30b-3p之间的靶向关系;3B双荧光素酶实验;3C RT-qPCR检测GABPB1-AS1和miR-30b-3p的表达。n=3;3B、3C两组间比较采用独立样本t检验。
图4 各组细胞活力情况比较注:CCK-8检测细胞活力(n=3)。多组间比较采用One-way ANOVA,Tukey's multiple comparisons test分析
图5 各组细胞凋亡情况比较注:流式细胞术检测细胞凋亡(n=3)。多组间比较采用One-way ANOVA,Tukey's multiple comparisons test分析
图6 各组细胞侵袭能力比较注:Transwell检测细胞侵袭(n=3)。多组间比较采用One-way ANOVA,Tukey's multiple comparisons test分析
图7 各组细胞迁移能力比较注:划痕试验检测细胞迁移(n=3)。多组间比较采用One-way ANOVA,Tukey's multiple comparisons test分析
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