Targeting granzyme B to tumor cells using a yoked human chorionic gonadotropin.

Autor: Kanatani I, Lin X, Yuan X, Manorek G, Shang X, Cheung LH, Rosenblum MG, Howell SB, Kanatani, Isao, Lin, Xinjian, Yuan, Xiaoqin, Manorek, Gerald, Shang, Xiying, Cheung, Lawrence H, Rosenblum, Michael G, Howell, Stephen B
Zdroj: Cancer Chemotherapy & Pharmacology; Oct2011, Vol. 68 Issue 4, p979-990, 12p
Abstrakt: Purpose: Luteinizing hormone receptor (LHR) is found in abundance on human ovarian, breast, endometrial and prostate carcinomas but at only low levels on non-gonadal tissues. To selectively kill LHR-expressing tumors, granzyme B (GrB) was linked to a protein in which both chains of human chorionic gonadotropin were yoked together (YCG).Methods: GrB-YCG was expressed and secreted from insect Sf9 cells. Its GrB enzymatic activity and binding affinity for hLHR were then characterized. The differential cytotoxicity of GrB-YCG versus GrB alone was tested in a panel of LHR-expressing tumor cells by SRB assay, and the mechanisms involved in the cell death were investigated by confocal fluorescence microscopy, flow cytometry, and western blot analysis.Results: GrB-YCG was successfully expressed and secreted from Sf9 insect cells and purified from cell culture supernatants. The serine protease activity of GrB-YCG was equivalent to that of human recombinant GrB. An in vitro hormone binding assay revealed that the GrB-YCG molecule also retained the ability to bind to the LHR receptor with an affinity similar to that of native hCG. Upon cell binding, GrB-YCG was rapidly internalized into LHR-expressing human ovarian cancer cells and produced selective and potent tumor cell killing by inducing apoptosis through activation of caspase-3.Conclusions: These results validate LHR as a therapeutic target and indicate that delivery of the human pro-apoptotic enzyme GrB to tumor cells by yoked hCG has substantial selectivity and therapeutic potential for human tumors that express high levels of LHR such as ovarian carcinomas. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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