MiR-125a-5p和信号传导与转录激活因子3在胃癌中的表达及其生物学功能

Expressions of miR-125a-5p, signal transducer and activator of transcription 3 in gastric cancer and their biological functions

  • 摘要:
      目的  采用生物信息学及分子生物学手段验证miR-125a-5p和信号传导与转录激活因子3(STAT3)在胃癌中的表达及其生物学功能。
      方法  从30例胃癌患者中分离出胃癌组织,采用实时荧光定量PCR(qRT-PCR)和蛋白质印迹(WB)法检测胃癌组织和非癌性胃组织中miR-125a-5p及STAT3的表达。细胞转染后,采用CCK-8检测细胞活力,流式细胞术检测细胞周期和凋亡情况。采用Transwell迁移实验检测迁移细胞。通过双荧光素酶报告基因检测验证miR-125a-5p与STAT3的靶向关系。通过体内裸鼠成瘤实验对miR-125a-5p的肿瘤抑制作用进行验证。
      结果  胃癌组织中miR-125a-5p表达较非癌性胃组织降低,差异有统计学意义(P < 0.05)。在胃癌细胞系中,与正常上皮细胞相比,miR-125a-5p的表达也下降,差异有统计学意义(P < 0.05)。采用miR-125a-5p模拟物转染胃癌细胞,结果表明miR-125a-5p的过表达可以通过靶向作用STAT3抑制胃癌细胞增殖、迁移和侵袭。裸鼠成瘤实验证实miR-125a-5p过表达能够抑制肿瘤增殖并促进肿瘤细胞凋亡。
      结论  MiR-125a-5p能够通过抑制STAT3的表达发挥肿瘤抑制作用,这将为临床靶向治疗及早期生物标志物的选择提供可靠的理论依据。

     

    Abstract:
      Objective  To verify the expression of miR-125a-5p, signal transducer and activator of transcription 3 (STAT3) in gastric cancer and their biological functions.
      Methods  Gastric cancer tissues were isolated from 30 patients with gastric cancer, and real-time fluorescent quantitative PCR (qRT-PCR) and Western blot (WB) were used to detect expression of miR-125a-5p and STAT3 in gastric cancer tissue and non-cancerous gastric tissue. After cell transfection, Cell Counting Kit-8 (CCK-8) was used to detect cell viability, and flow cytometry was used to detect cell cycle and apoptosis. Transwell migration test was used to detect migrating cells. The target relationship between miR-125a-5p and STAT3 was verified by dual luciferase reporter gene detection. The tumor inhibitory effect of miR-125a-5p was verified by tumor formation experiment in nude mice in vivo.
      Results  The expression of miR-125a-5p in gastric cancer tissue was significantly lower than that in non-cancerous gastric tissue (P < 0.05). In gastric cancer cell lines, the expression of miR-125a-5p was also significantly lower than that in the normal epithelial cells (P < 0.05). In addition, authors transfected gastric cancer cells with miR-125a-5p mimics, and the results showed that the over-expression of miR-125a-5p was able to inhibit the proliferation, migration and invasion of gastric cancer cells by targeting STAT3. Tumor formation experiment in nude mice verified that over-expression of miR-125a-5p was able to inhibit tumor proliferation and promote tumor cell apoptosis.
      Conclusion  MiR-125a-5p can inhibit tumors by inhibiting the expression of STAT3, and these findings will provide reliable theoretical bases for clinical targeted therapy and early selection of biomarkers.

     

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