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Objective: To investigate the role of tissue factor (TF) in chemotherapeutic reagent - induced apoptosis on human glioblastoma and explore its mechanism. Methods: The expression of TF was examined by Western blotting. The cytotoxicity of doxorubicin was determined by WST assay. The activation of Caspase-3 and PARP induced by adoxorubicin were tested by Western blotting. Results: Human glioblastoma cell line U373MG expressed high level of TF while LN-229 was with low-TF level. The chemotherapeutic reagent doxorubicin revealed stronger cytotoxic effect on high-TF U373MG cells than low-TF LN-229 cells. Enforced strong expression of TF was achieved by transfection of TF-pcDNA3 combinant on LN-229 cells in a dose-dependent manner. Enforced TF expression in transfected LN-229 cells not only impaired the doxorubicin-induced cleavage of Caspase-3 and PARP, but also inhibited the cytotoxic effect of doxorubicin. Furthermore, activation of Akt was strong in high-TF U373MG cells but weak in low-TF LN-229 cells. Incubation of factor VII (FVII) with enforced TF-expressing LN-229 cells increased the phosphorylation of Akt in a time-dependent manner. Conclusion: These results suggest that over-expression of TF on glioblastoma could inhibit doxorubicin-induced apoptosis. Interaction of FVII and TF activates the downstream PI3K/Akt pathway. Tumor-derived over-expression of TF might play a role in chemotherapy resistance in glioblastoma, at lest in part, by activating PI3K/Akt-mediated survival and anti-apoptotic mechanism through the interaction of TF/FVII signaling.
Objective: To investigate the role of tissue factor (TF) in chemotherapeutic reagent - induced apoptosis on human glioblastoma and explore its mechanism. Methods: The expression of TF was examined by Western blotting. The cytotoxicity of doxorubicin was determined by WST assay. The activation of Caspase-3 and PARP induced by adoxorubicin were tested by Western blotting. Results: Human glioblastoma cell line U373MG expressed high level of TF while LN-229 was with low-TF level. The chemotherapeutic reagent doxorubicin revealed stronger cytotoxic effect on high-TF Enforced TF expression in transfected LN-229 cells not only impaired the U373MG cells than low-TF LN-229 cells. Enforced TF expression achieved TF was achieved by transfection of TF-pcDNA3 combinant on LN-229 cells in a dose- dependent manner. doxorubicin-induced cleavage of Caspase-3 and PARP, but also inhibited the cytotoxic effect of doxorubicin. Furthermore, activation of Akt was strong in high-TF U373MG cells but weak in low-T Incubation of factor VII (FVII) with enforced TF-expressing LN-229 cells increased the phosphorylation of Akt in a time-dependent manner. Conclusion: These results suggest that over-expression of TF on glioblastoma could inhibit doxorubicin -induced apoptosis. Interaction of FVII and TF activates the downstream PI3K / Akt pathway. Tumor-derived over-expression of TF might play a role in chemotherapy resistance in glioblastoma, at lest in part, by activating PI3K / Akt-mediated survival and anti -apoptotic mechanism through the interaction of TF / FVII signaling.