金桃花, 雷鑫明, 童秀萍. 长链非编码RNA AGAP2-AS1通过YAP通路影响结肠癌细胞的生物学行为[J]. 中国肿瘤临床, 2020, 47(12): 601-608. DOI: 10.3969/j.issn.1000-8179.2020.12.403
引用本文: 金桃花, 雷鑫明, 童秀萍. 长链非编码RNA AGAP2-AS1通过YAP通路影响结肠癌细胞的生物学行为[J]. 中国肿瘤临床, 2020, 47(12): 601-608. DOI: 10.3969/j.issn.1000-8179.2020.12.403
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Taohua Jin, Xinming Lei, Xiuping Tong. Long non-coding RNA AGAP2-AS1 affects biological behavior of colon cancer cells through the YAP signaling pathway[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2020, 47(12): 601-608. DOI: 10.3969/j.issn.1000-8179.2020.12.403
Citation: Taohua Jin, Xinming Lei, Xiuping Tong. Long non-coding RNA AGAP2-AS1 affects biological behavior of colon cancer cells through the YAP signaling pathway[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2020, 47(12): 601-608. DOI: 10.3969/j.issn.1000-8179.2020.12.403
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长链非编码RNA AGAP2-AS1通过YAP通路影响结肠癌细胞的生物学行为

Long non-coding RNA AGAP2-AS1 affects biological behavior of colon cancer cells through the YAP signaling pathway

    摘要
  • 摘要:
      目的  探究AGAP2-AS1在结肠癌组织中的表达及对结肠癌恶性生物学行为和Yes相关蛋白（Yes-associated protein，YAP）信号通路的影响。
      方法  通过TCGA数据库分析457例结肠癌和42例健康样本的AGAP2-AS1表达水平。收集于2018年1月至2019年3月在义乌市中心医院就诊且通过病理科确诊的结肠癌组织和癌旁组织20例，通过实时荧光定量PCR检测其AGAP2-AS1的表达，之后进一步检测SW480、HCT-116和NCM460细胞中的AGAP2-AS1表达。利用CCK-8试剂盒、克隆形成、细胞划痕和流式细胞实验分析其细胞增殖、迁移、侵袭及凋亡的改变。Western blot检测YAP、p-YAP以及基质金属蛋白酶（matrix metallo-peptidase，MMP）2、MMP9的表达。免疫荧光检测YAP在细胞内的定位情况。共转染pcDNA3.1-AGAP2-AS1和si-YAP质粒，然后检测YAP、p-YAP、MMP2、MMP9蛋白的表达。
      结果  TCGA数据库显示AGAP2-AS1在结肠癌组织显著高表达，并且AGAP2-AS1在结肠癌组织样本和细胞中表达也显著增加。敲低/过表达AGAP2-AS1后HCT-116细胞迁移，增殖能力显著降低/增加，凋亡显著增加/抑制；同时敲低AGAP2-AS1后会诱导YAP磷酸化，减少YAP入核调控，且MMP2、MMP9的表达也显著降低（P < 0.05）。共转染结果显示AGAP2-AS1通过激活YAP通路上调MMP2、MMP9表达（P < 0.05）。
      结论  AGAP2-AS1在结肠癌组织和细胞系中均高表达，且诱导结肠癌细胞增殖、迁移并抑制细胞凋亡。此外，AGAP2-AS1通过激活YAP通路调控MMP2、MMP9的表达影响结肠癌侵袭和转移。

     

    Abstract:
      Objective  To investigate the expression of AGAP2-AS1 in colon cancer, its influence on the malignant behavior of colon cancer cells, and its effects on the Yes-associated protein (YAP) signaling pathway.
      Methods  The expression of AGAP2-AS1 in 457 colon cancer samples and 42 healthy samples was obtained from TCGA. Twenty matched colon cancer and adjacent cancer tissues were collected from patients diagnosed in Yiwu Central Hospital and confirmed by pathology department from January 2018 to March 2019. The expression of AGAP2-AS1 in these tissues, as well as in SW480, HCT-116, and NCM460 cells, was detected by real-time fluorescent quantitative PCR. The changes in cell proliferation, migration and apoptosis were analyzed and detected using a CCK-8 kit, and clone formation, wound-healing assay, and flow cytometry experiments. The levels of YAP, p-YAP, matrix metalloproteinase-2 (MMP2), and MMP9 were evaluated by Western blot. Immunofluorescence was used to examine the localization of YAP in HCT-116 cells. The levels of YAP, p-YAP, MMP2, and MMP9 proteins were evaluated after the co-transfection of pcDNA3.1-AGAP2-AS1 and si-YAP plasmids.
      Results  TCGA indicated that the expression of AGAP2-AS1 was significantly increased in colon cancer tissues. Similarly, in the present study, AGAP2-AS1 expression was significantly increased in colon cancer tissues and cells. After AGAP2-AS1 knockdown/overexpression, the capacity of migration and proliferation was significantly reduced/increased, and apoptosis was significantly increased/inhibited in colon cancer cells. Moreover, AGAP2-AS1 knockdown triggered the phosphorylation of YAP, and a significant reduction in the expression of MMP2 and MMP9 (P < 0.05). Co-transfection studies revealed that AGAP2-AS1 upregulated the expression of MMP2 and MMP9 via activation of the YAP pathway (P < 0.05).
      Conclusion  AGAP2-AS1 was highly expressed in colon cancer tissues and cell lines. AGAP2-AS1 can induce proliferation and migration, and inhibit apoptosis in colon cancer cells. In addition, AGAP2-AS1 regulated MMP2 and MMP9 expression by activating the YAP pathway, thereby affecting colon cancer metastasis.

     

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