Gastrointestinal Unit, Mater Misericordiae University Hospital, Eccles Street, Dublin 7, Ireland.
J Exp Clin Cancer Res. 2010 Jun 22;29(1):81. doi: 10.1186/1756-9966-29-81.
We have previously reported that Myeov (MYEloma OVerexpressed gene) expression is enhanced in colorectal cancer (CRC) and that it promotes CRC cell proliferation and invasion. The role of Myeov in CRC migration is unclear. ProstaglandinE2 (PGE 2) is a known factor in promoting CRC carcinogenesis. The role of PGE 2 in modulating Myeov expression has also not been defined.
To assess the role of Myeov expression in CRC cell migration and to evaluate the role of PGE 2 in Myeov bioactivity.
siRNA mediated Myeov knockdown was achieved in T84 CRC cells. Knockdown was assessed using quantitative real time PCR. The effect of knockdown on CRC cell migration was assessed using a scratch wound healing assay. Separately, T84 cells were treated with PGE 2 (0.00025 micro M, 0.1 micro M and 1 micro M) from 30 min to 3 hours and the effect on Myeov gene expression was assessed using real time PCR.
Myeov knockdown resulted in a significant reduction in CRC cell migration, observable as early as 12 hours (P < 0.05) with a 39% reduction compared to control at 36 hours (p < 0.01). Myeov expression was enhanced after treatment with PGE 2, with the greatest effect seen at 60 mins for all 3 PGE 2 doses. This response was dose dependent with a 290%, 550% & 1,000% increase in Myeov expression for 0.00025 micro M, 0.1 micro M and 1 micro M PGE 2 respectively.
In addition to promoting CRC proliferation and invasion, our findings indicate that Myeov stimulates CRC cell migration, and its expression may be PGE 2 dependant.
我们之前曾报道过，Myeov（骨髓瘤过度表达基因）在结直肠癌（CRC）中表达增强，并促进 CRC 细胞增殖和侵袭。Myeov 在 CRC 迁移中的作用尚不清楚。前列腺素 E2（PGE2）是促进 CRC 癌变的已知因素。PGE2 调节 Myeov 表达的作用也尚未确定。
评估 Myeov 表达在 CRC 细胞迁移中的作用，并评估 PGE2 在 Myeov 生物活性中的作用。
采用 siRNA 介导的 Myeov 敲低方法在 T84 CRC 细胞中实现。通过定量实时 PCR 评估敲低情况。通过划痕愈合实验评估敲低对 CRC 细胞迁移的影响。另外，T84 细胞分别用 PGE2（0.00025 μM、0.1 μM 和 1 μM）处理 30 分钟至 3 小时，通过实时 PCR 评估对 Myeov 基因表达的影响。
Myeov 敲低导致 CRC 细胞迁移明显减少，在 12 小时（P < 0.05）即可观察到，在 36 小时（p < 0.01）与对照组相比降低 39%。PGE2 处理后 Myeov 表达增强，所有 3 种 PGE2 剂量的最大效应在 60 分钟时出现。这种反应呈剂量依赖性，0.00025 μM、0.1 μM 和 1 μM PGE2 分别使 Myeov 表达增加 290%、550%和 1000%。
除了促进 CRC 增殖和侵袭外，我们的研究结果表明，Myeov 刺激 CRC 细胞迁移，其表达可能依赖于 PGE2。
