HCT-116 colorectal cancer cells secrete chemokines which induce chemoattraction and intracellular calcium mobilization in NK92 cells
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We recently reported that pretreatment of IL-2 activated human natural killer (NK) cells with the drugs dimethyl fumarate (DMF) and monomethyl fumarate (MMF) upregulated the expression of surface chemokine receptor CCR10. Ligands for CCR10, namely CCL27 and CCL28, induced the chemotaxis of these cells. Here, we performed a bioinformatics analysis to see which chemokines might be expressed by the human HCT-116 colorectal cancer cells. We observed that, in addition to CCL27 and CCL28, HCT-116 colorectal cancer cells profoundly express CXCL16 which binds CXCR6. Consequently, NK92 cells were treated with DMF and MMF for 24 h to investigate in vitro chemotaxis towards CXCL16, CCL27, and CCL28. Furthermore, supernatants collected from HCT-116 cells after 24 or 48 h incubation induced the chemotaxis of NK92 cells. Similar to their effects on human IL-2-activated NK cells, MMF and DMF enhanced the expression of CCR10 and CXCR6 in NK92 cells. Neutralizing anti-CXCL16 or anti-CCL28 inhibited the chemotactic effects of 24 and 48 supernatants, whereas anti-CCL27 only inhibited the 48 h supernatant activity, suggesting that 24 h supernatant contains CXCL16 and CCL28, whereas HCT-116 secretes all three chemokines after 48 h in vitro cultures. CXCL16, CCL27, and CCL28, as well as the supernatants collected from HCT-116, induced the mobilization of (Ca)2+ in NK92 cells. Cross-desensitization experiments confirmed the results of the chemotaxis experiments. Finally, incubation of NK92 cells with HCT-116 induced the lysis of the tumor cells. In summary, these results might have important implications in directing the anti-tumor effectors NK cells towards tumor growth sites.
KeywordsColorectal cancer NK92 cells Chemokines Dimethyl fumarate Monomethyl fumarate Calcium mobilization
CC chemokine ligand/chemokine receptor
Cutaneous T-cell-attracting chemokine/CCL27
CXC chemokine ligand/chemokine receptor
Experimental autoimmune encephalomyelitis
Ethylene glycol-bis(2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid
Enzyme-linked immunosorbent assay
Program death ligand-1
Real-time quantitative polymerase chain reaction
Roswell Park Memorial Institute
Regulatory T cells
NME performed most of the experiments and wrote the paper; ZAJ performed the calcium assays; MYH performed the bioinformatics and qRT-PCR; AAM designed the experiments, performed the statistical analysis, and wrote the manuscript.
This work was supported by the University of Sharjah Grants with numbers 1701090222-P and 1701090223-P.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The work described in this paper was performed using commercially available cell lines. This article does not contain any studies involving patients or experimental animals. Therefore, no study approval was required and no informed consent from the donors.
Cell line authentication
The human natural killer cell line NK92 (CRL-2407), colorectal cancer cell line HCT-116 (CCL-247), and the erythroleukemia K562 (CCL-243) were obtained from the American type culture collection (ATCC, Manassas, VA, USA). No cell line authentication was necessary.
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