Establishment of multidrug resistance cell line of cisplatin and gemcitabine in bladder cancer cell line T24 and its biological characteristics

Abstract

Objective To establish a human bladder cancer cisplatin (DDP) and gemcitabine (GEM) resistant cell line (T24/DDP&GEM) by concentration gradient method, and to observe its biological characteristics. Methods DDP and GEM were used as inducers to establish human bladder cancer cisplatin and gemcitabine resistant model T24/DDP&GEM in vitro. The morphology of T24 and T24/DDP&GEM cells was observed by the inverted microscope. The drug resistance and the growth curve of cells were detected by cell counting kit-8 (CCK-8). The P-glycoprotein (P-gP) expression was detected by Western blotting. Results After the induction of DDP and GEM for 12 months, T24/DDP&GEM was successfully established. Under the inverted phase contrast microscopy, as compared with T24 cells, the morphology of T24DDP/GEM cells was irregular, the volume became larger, and vacuoles and granules were formed in the cytoplasm. The proliferation ability of T24/DDP&GEM cells was significantly lower than that of T24 cells. The doubling time of T24/DDP&GEM cells was (39.51±0.36) h, and that of T24 was (28.06±0.76) h. The doubling time was extended for (11.45±0.41) h. The drug resistance of T24/DDP&GEM was significantly improved, and the drug resistance index was 4.25 for DDP and 62.97 for GEM. As compared with T24 cells, the proportion of S and G2 phase cells in T24/DDP&GEM cells decreased, the proportion of G1 cells increased significantly, and the cell cycle slowed down. The results of Western blotting showed that the expression level of P-gp in T24/DDP&GEM cells was significantly higher than that in T24 cells. Conclusion The artificial induced DDP and GEM resistant bladder cancer cell line T24/DDP&GEM has the characteristics of multi drug resistant tumor cells. Key words: Bladder cancer; Chemotherapy; Cisplatin; Gemcitabine; Multidrug resistance