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Associations of MICA gene polymorphism with susceptibility of breast cancer cells to NK cell-mediated cytotoxicity
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Abstract:
Objective: To investigate the associations of MHC class I chain-related molecule A（MICA）gene polymorphism with susceptibility of breast cancer cells to NK cell-mediated cytotoxicity. Methods: MICA alleles of breast cancer cell lines（MCF-7, MDAMB-231, MDA-MB-435S and SK-BR-3）were analyzed by DNA sequencing. MICA protein expression in 293T cells transfected with MICA recombinant expression vectors (namely pMCFA5.1 ，pMCFA4 ，p231A5R ，p231A9 and p435A5P) was detected by Western blotting and Flow cytometry; the cytotoxicity of NK cells against the above 293T cells were measured by lactate dehydrogenase （LDH）assay and the release of perforin（PFN）and granzymes B（Gzm B）were measured by ELISPOT assay. Results: DNA sequencing result showed that MICA*008/A5.1 and MICA*001/A4 were expressed in MCF-7 cells, MICA*019/A5 and MICA*002/A9 were expressed in both MDA-MB-231 and SK-BR-3 cells while MICA*010/A5 was expressed in MDA-MB-435S cells. After the MICA recombinant expression vectors were transfected into 293T cells, the level of MICA were the lowest in pMCFA5.1 group (P<0.05), following after p435A5P group (P<0.05), and highly expressed in the pMCFA4, p231AR and p231A9 groups (P<0.05). NK cell-mediated cytotoxicity and the release of PFN and Gzm B: NK cell-mediated cytotoxicity were the lowest in p435A5P group (P<0.05), and the ability of inducing NK cells to release PFN and Gzm B was also the lowest in p435A5P group (P<0.05), but there were no statistical difference among the other transfected groups (P>0.05). Conclusion: MICA gene polymorphism is closely associated with susceptibility of breast cancer cells to NK cell-mediated cytotoxicity.
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Project Supported:
Project supported by the National Clinical Key Specialty Construction Program of China（Letter of the National Health Office [2013] 544），and the Natural Science Foundation of Fujian Province（No. 2015J01433）
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