Intercellular Selectively Transfer Of P-glycoprotein Between Different Bladder Cancer Cell Lines

Objective:To investigate the effects of P-gp and multidrug resistance gene(MDR1) transfer, whether the functional P- glycoprotein in different cell lines of bladder cancer and the relationship between glycoprotein transfer selective mechanism.Methods: To coculture BIU-87/ADM cells of bladder cancer resistant cells with T24 and EJ sensitive bladder cancer cell lines use Transwell. Confocal microscopy to observe transfer of P- glycoprotein expression of T24 and EJ cells which after co-culture with BIU-87/ADM. Using the Western Blot analyze the P- glycoprotein expression of BIU-87/ADM、T24/ aq MDR、T24、EJ/ aq MDR and EJ cells.Using RT-PCR analyze the expression levels of MDR1 m RNA in these cells. Measured fluorescence intensity of rhodamine-123 in BIU-87/ADM、T24/ aq MDR、T24、 EJ/ aq MDR and EJ cells by flow cytometry. To use RT-PCR detected the expression of m RNA of CD44 、 Ezrin and integrin b1 protein molecule in BIU-87 、 EJ 、BIU-87/ADM、T24 cells.Results:The results showed that the confocal microscopy showed nucleus is blue fluorescence, the cytoplasm and membrane showed green fluorescence in different degree, BIU-87/ADM showed strong green fluorescence, T24/ aq MDR showed moderate intensity of green fluorescence.while T24、EJ/ aq MDR and EJ cells showed a weak green fluorescence. Western Blot results showed that the highest level of expression of P-gp in BIU-87/ADM cells.T24/ aq MDR cells in the expression of P-gp was weaker than BIU-87/ADM cells(p<0.05) but stronger than that of T24 cells(p<0.05).But EJ/ aq MDR and EJ cells in the expression level of P-gp was lower, There was no significant difference between the them. RT-PCR results showed thatonly MDR1 m RNA levels are higher in BIU-87/ADM cell, while The rest of the four types of cells expression level is very low, there was no significant difference(p>0.05). The Rhodmine 123 test showed that the fluorescence intensity of BIU-87/ADM、 T24/aq MDR、T24、EJ/aq MDR and EJ cells were increased gradually. The T24, EJ/aq MDR and EJ cell fluorescence intensity is strong, there is no statistically significant difference between each other(p>0.05). The expression level of CD44 gene detection、Ezrin protein、integrin b1 receptor were high to show in seven kinds of cells, but also no statistically significant difference between each other(p>0.05).Conclusion: P-gp can be transferred from resistant human bladder cancer cell line BIU-87/ADM of the sensitivity of the bladder cancer cell line T24, which acquired P-gp with drug efflux function. But P-gp can not or very little from the resistant human bladder cancer cell line BIU-87/ADM to bladder cancer cells in EJ sensitive. In the coculture of P-gp before and after the transfer of the transfer process is not associated with multidrug resistance gene MDR1. This finding indicates that the transfer of P-gp in bladder cancer cell line transfer is selective which by Membrane microparticles(MPs) mediated mechanism. The expression of CD44、Ezrin、integrin ?1 receptors in all cells have no significant difference. No sufficient evidence to show the protein take part in the transfer of P-gp. It provides a new thought and a new research direction for further study on the mechanism of multidrug resistance mechanism by P-gp mediated in bladder cancer.