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LncRNA TBX5-AS1靶向miR-92a-3p调控结直肠癌HCT116细胞增殖、迁移及侵袭的机制研究

Study on mechanism of LncRNA TBX5-AS1 in regulating proliferation, migration and invasion of colorectal cancer HCT116 cells by targeting miR-92a-3p

    摘要
  • 摘要:
      目的  探讨长链非编码RNA T-box转录因子5反义RNA 1(LncRNA TBX5-AS1)对结直肠癌细胞生物学行为的影响及其可能作用机制。
      方法  实时荧光定量聚合酶链反应(qRT-PCR)法检测结直肠癌组织、癌旁组织中TBX5-AS1和miR-92a-3p的表达量。体外培养人结直肠癌细胞HCT116, 分别将TBX5-AS1过表达载体(pcDNA-TBX5-AS1)及其对照空载体(pcDNA)、miR-92a-3p特异性寡核苷酸抑制剂(anti-miR-92a-3p)及其阴性对照(anti-miR-NC)、miR-92a-3p寡核苷酸模拟物(miR-92a-3p mmics)及阴性对照mimic NC序列(miR-NC)、pcDNA-TBX5-AS1与miR-92a-3p mimics转染至HCT116细胞。MTT法、平板克隆形成实验、划痕实验与Transwell小室实验分别检测细胞增殖、克隆形成、迁移及侵袭能力; 双荧光素酶报告基因实验检测TBX5-AS1和miR-92a-3p的靶向关系。
      结果  与癌旁组织比较，结直肠癌组织中TBX5-AS1的表达量降低, miR-92a-3p的表达量升高，差异有统计学意义(P < 0.05)。转染pcDNA-TBX5-AS1或转染anti-miR-92a-3p后细胞活力降低，克隆形成数和侵袭细胞数减少，迁移距离缩短，差异均有统计学意义(P < 0.05)。TBX5-AS1可靶向调控miR-92a-3p; 转染miR-92a-3p mmics后细胞活力升高，克隆形成数和侵袭细胞数增多，迁移距离增加，差异有统计学意义(P < 0.05)。共转染pcDNA-TBX5-AS1与miR-92a-3p mmics能够逆转pcDNA-TBX5-AS1对HCT116细胞生物学行为的作用。
      结论  TBX5-AS1过表达可通过抑制miR-92a-3p的表达，减弱结直肠癌细胞增殖、克隆形成、迁移及侵袭能力。

     

    Abstract:
      Objective  To explore the effect of long non-coding RNA T-box transcription factor 5 antisense RNA 1 (LncRNA TBX5-AS1) on the biological behavior of colorectal cancer cells and its possible mechanism.
      Methods  Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) method was used to detect the expression of TBX5-AS1 and miR-92a-3p in colorectal cancer tissues and adjacent tissues. Human colorectal cancer cell HCT116 was cultured in vitro, and TBX5-AS1 overexpression vector (pcDNA-TBX5-AS1) and its control empty vector (pcDNA), miR-92a-3p specific oligonucleotide inhibitor (anti-miR-92a-3p) and its negative control (anti-miR-NC), miR-92a-3p oligonucleotide mimics (miR-92a-3p mmics) and negative control mimic NC sequence (miR-NC), pcDNA-TBX5-AS1 and miR-92a-3p mimics were transfected into HCT116 cells. MTT method, plate colony formation experiment, scratch experiment and Transwell chamber experiment were used to detect cell proliferation, colony formation, migration and invasion ability respectively. The dual luciferase reporter gene experiment was used to detect the targeting relationship between TBX5-AS1 and miR-92a-3p.
      Results  Compared with adjacent tissues, the expression of TBX5-AS1 in colorectal cancer tissues decreased significantly, while the expression of miR-92a-3p increased significantly (P < 0.05). After transfection with pcDNA-TBX5-AS1 or anti-miR-92a-3p, cell viability decreased significantly, the number of colony formation and invasive cells decreased significantly, and the migration distance shortened significantly (P < 0.05). TBX5-AS1 was able to target miR-92a-3p. After transfection with miR-92a-3p mimics, cell viability increased significantly, the number of colony formation and invasive cells increased significantly, and the migration distance increased significantly (P < 0.05). Co-transfection with pcDNA-TBX5-AS1 and miR-92a-3p mmics was able to reverse the effect of pcDNA-TBX5-AS1 on the biological behavior of HCT116 cells.
      Conclusion  Overexpression of TBX5-AS1 can reduce the proliferation, colony formation, migration and invasion abilities of colorectal cancer cells by inhibiting the expression of miR-92a-3p.

     

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