Construction of Trop-2-targeted chimeric antigen receptor-modified T cells and their effects on the proliferation of ovarian cancer cells in vitro
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    Abstract:

    Objective: To produce Trop-2-targeted chimeric antigen receptor T cells(CAR-T cells) of human trophoblast cell surface antigen 2(Trop-2) and detect their effects on the proliferation of ovarian cancer cells in vitro. Methods: Molecular cloning and gene recombination technology were introduced to construct Trop-2 CAR vector. The expression of Trop-2 CAR on 293T cells was tested by Western blot. The effect of Trop-2 CAR-T cells on the proliferation of ovarian cancer cells was evaluated by CCK-8 assay. The secreting changes of cytokines by Trop-2 CAR-T cells were detected by ELISA. Results: The results of enzyme digestion and sequencing demonstrated that the Trop-2 CAR vector was constructed correctly. The results of Western blot showed that Trop-2 CAR was expressed on 293T cells. The results of CCK-8 verified that Trop-2 CAR-T cells prominently inhibited the growth of ovarian cancer cells with Trop-2 antigen expression(P<0.05), moreover, higher levels of IFN-γ and IL-2 were detected in supernatant by ELISA(P<0.01). Conclusion: Trop-2 CAR-T cells were prepared successfully and could inhibit the proliferation of ovarian cancer cells expressing Trop-2 antigen.

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徐亚如,周 荧,唐 奇,刘振云,黄骁辰,杨婷婷,张慧林,赵 薇,蒯兴旺,仇镇宁,朱 进,冯振卿.靶向Trop-2 CAR-T细胞的制备及其体外对卵巢癌细胞杀伤作用的研究[J].南京医科大学学报(自然科学版英文版),2017,(6):653-658.

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  • Received:January 12,2017
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  • Online: June 30,2017
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