TRPV5 TRPV6对结肠癌SW480细胞生物学行为的影响

Effect of TRPV5 and TRPV6 channels on the biological behaviors of SW480 colon cancer cell line

    摘要
  • 摘要:
      目的  观察TRPV5和TRPV6蛋白表达水平对结肠癌SW480细胞内Ca2+水平及细胞增殖、凋亡、迁移等生物学行为的影响。
      方法  给予TRPV5、TRPV6通道激动剂1-25（OH）2D3及抑制剂CuCl2作用结肠癌SW480细胞后，采用免疫组织化学法、Western blot及实时荧光定量PCR技术检测TRPV5、TRPV6蛋白及TRPV5、TRPV6 mRNA表达的变化。使用高速离子成像系统观察结肠癌SW480细胞内Ca2+水平的变化；通过MTT法、TUNEL法及划痕修复法观察结肠癌SW480细胞增殖凋亡及迁移的变化。
      结果  1-25（OH）2D3明显上调结肠癌SW480细胞TRPV5、TRPV6 mRNA及蛋白表达水平，使结肠癌SW480细胞内Ca2+水平增加，促进结肠癌SW480细胞增殖、迁移及抑制其凋亡（P＜0.05）。CuCl2明显下调结肠癌SW480细胞中TRPV5、TRPV6蛋白及mRNA表达，使结肠癌SW480细胞内Ca2+水平降低（P＜0.05），抑制结肠癌SW480细胞的增殖、迁移及促进细胞的凋亡（P＜0.05）。
      结论  结肠癌SW480细胞中TRPV5和TRPV6蛋白表达水平的变化，能通过调控细胞内Ca2+水平影响细胞的增殖、迁移和凋亡等生物学行为。

     

    Abstract:
      Objective  To investigate the role of TRPV5 and TRPV6 in intracellular calcium regulation and biological behaviors of SW480colon cancer cells.
      Methods  qRT-PCR, Western blot, and immunocytochemistry were applied to determine the mRNA and protein expression levels of TRPV5 and TRPV6 in SW480 colon cancer cell line upon treatment with TRPV5 and TRPV6 agonist, 1-25(OH)2D3, and inhibitor, CuCl2. The change of intracellular Ca2+ level was examined with a confocal laser scanning microscope. Scratch test, MTT, and TUNEL assays were used to analyze the cell migration, proliferation, and apoptosis, respectively.
      Results  As an agonist of TRPV5 andTRPV6, 1-25(OH)2D3 significantly up-regulated the mRNA and protein expression levels of TRPV5 and TRPV6 in SW480 cell lines. On the other hand, CuCl2, being an inhibitor of TRPV5 and TRPV6, effectively down-regulated the TRPV5 and TRPV6 mRNA and protein expression levels (P < 0.05). The intracellular calcium concentration in SW480 cell line significantly increased upon treatment with 1-25 (OH)2D3, and significantly decreased with CuCl2 treatment (P < 0.05). 1-25(OH)2D3 promoted cell proliferation and migration, and inhibited apoptosis of SW480 cell in a time-and dose-dependent manner (P < 0.05). However, CuCl2 significantly repressed cell proliferation and migration and induced apoptosis (P < 0.05).
      Conclusion  TRPV5 and TRPV6 can affect the biological behaviors of colon cancerSW480 cells by regulating intracellular Ca2+ level.

     

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