ABSTRACT
Aim The TRPV1 plasmid was transiently transfected into human embryonic kidney HEK 293T cells to establish the heterologous expression system of TRPV1-channel. Methods The transfection efficiency was confirmed under fluorescence mi-croscope and the TRPV1 protein expression was identified by u-sing Western blot, and the functional characteristics of the chan-nel were studied by using the method of confocal microscopy. Results The transfection rate could reach 40% ~50%; the transfected cells were found to have a clear band at the corre-sponding position that TRPV1 should be, which indicated that TRPV1 channel protein was expressed in the transfected cells. The confocal microscopy imaging result showed that the trans-fected HEK 293T cells were activated by TRPV1 channel ago-nist. Conclusion Transient transfection of HEK 293T cells with TRPV1 channel is successfully constructed and the heterol-ogous TRPV1 channel is verified to have normal calcium-media-ting function.
ABSTRACT
Objective: To study the effect of TIP30 gene on the cell cy cle distribution in adenoid cystic carcinoma ACC-M cells. Methods: Wild type TIP30 gene was transfected into ACC-M cells by using lipofectamin e, the postive cell clone was selected with G418,the expression of TIP30 protei n in the cells was detected by Western blot. Cell cycle distribution was detecte d by flow cytometry. Results:14 days after seletion anti-G418 c ell clone was obtained. Wstern blot revealed that the TIP30 protein was expresse d in the transfected cells.Flow cytometric analysis indicated that TIP30 induced a arrest of the cells in G 0-G 1 phase and led to a decrease of cell percent age in G 2-M and S phase. Conclusion:ACC-M can stably express exogene TIP30.TIP30 can inhibit proliferation of ACC-M cells.