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Archive > Volume 21 Issue 3 > 2014,21(3):263-268. DOI:10.3872/j.issn.1007-385X.2014.03.005 Prev Next
Basic Research
The inhibitory effect of hCaMKⅡNα on the production of immunosuppressive factors in colon cancer cells in vitro
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Abstract:
To investigate the effect of (human calcium/calmodulin-dependent protein kinase Ⅱ inhibitory alpha (hCaMKⅡN-α) on the production of immunosuppressive factors in colon cancer cells and the mechanisms underlying the effect in vitro. Methods: Overexpression and silencing of the hCaMKⅡN-α gene in human colon adenocarcinoma (LoVo, SW620 and HT29) cells were achieved by transfection with a hCaMKⅡN-α-expressing plasmid (pKⅡN-α) and an siRNA (si-KⅡN-α ) vector, respectively. Messenger RNA levels of interleukin-8 (IL-8), interleukin-10 (IL-10) and vascular endothelial cell growth factor (VEGF) in LoVo cells transfected with pKⅡN- were analyzed by RT-PCR. Protein levels of IL-8, IL-10 and VEGF in SW620 and LoVo cells transfected with pKⅡN- and in HT29 cells transfected with si-KⅡN- were determined by ELISA. The differences in IL-8 and VEGF protein levels in HT29 cells transfected with pKIN-α in the presence or absence of U0126 (10 M), a selective ERK1/2 inhibitor, were analyzed to elucidate the role of ERK1/2 in hCaMKⅡN-α-mediated IL-8 and VEGF production. Results: Overexpression of hCaMKⅡN- significantly decreased the mRNA abundance and protein levels of VEGF and IL-8 (P<0.05) but not PGE2 (P>0.01). Silencing of hCaMKⅡN- by siRNA significantly increased the secretion of VEGF and IL-8 in HT29 cells, but had no effect on the secretion of PGE2 and IL-10. U0126 treatment resulted in a complete reversion of increased IL-8 secretion but only a partial reversion of increased VEGF secretion in HT29 cells overexpressing hCaMKⅡ-α. Conclusion: Our observations suggest that hCaMKⅡ- may inhibit the secretion of VEGF and IL-8 and thus down-regulate the immune response in rectal tumor cells through an ERK signaling pathway-dependent mechanism.
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Project Supported:
Project supported by the National Natural Secience Foundation of China（No. 31270931）
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