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Archive > Volume 23 Issue 5 > 2016,23(5):620-625. DOI:10.3872/j.issn.1007-385X.2016.05.006 Prev Next
Basic Research
Killing effect of NK-92MI cells modified with chimeric antigen receptor on HER2 + breast carcinoma cell
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Abstract:
Objective:To explore killing efficiency of NK-92MI cells modified with chimeric antigen receptor of specific targeting HER2 antigen (HER2-CAR) on HER2+ breast carcinoma cells. Methods: HER2-CAR fragments were obtained by PCR, then built into lentiviral vectors via molecular cloning technology and transfected into NK-92MI cells using the lentiviral particles with HER2-CAR. Flow cytometry assay was used to detect efficiency of the cell transfection and, secretion differences between IFN-γ and granzyme in NK-92MI cells before and after the transfection. Killing efficiency of HER2-CAR-NK92MI cells on breast carcinoma cells in vitro was tested with 7-AAD and flow cytometry assays. Mouse model with breast tumor in situ was constructed and used to detect killing efficiency of the HER2-CAR-NK92MI cells in vivo. Results:Killing efficiencies of the NK-92MI cell modified with HER2-CAR on HER2+ breast carcinoma MCF-7 and T47T cell lines were significantly higher than those of the NK-92MI cell that did not modified with HER2-CAR gene (\[62.4±1.3\]% vs \[22.1±1.2\]%. \[50.0±1.9\]% vs \[16.9±0.6\]%， respectively, all P<0.01). However, there was no obvious difference in killing efficiency of both the modified and un-modified NK-92MI cells on HER2- breast carcinoma MDA-MB-468 cell (\[13.6±1.4\]% vs \[12.7±0.8\]%,P>0.05). At the same time, amount of IFN-γ secreted by the NK-92MI cell modified with HER2-CAR was significantly higher than that secreted by the NK-92MI cell which did not modified with HER2-CAR gene (P<0.01).Conclusion: Killing efficiency of the NK-92MI cell modified with HER2-CAR on HER2+ breast carcinoma cell significantly increased, and could have specific targeting property, which might provide a clinical evidence for the treatment of malignant tumor, such as breast carcinoma and so on.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No.31471283）
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