Objective:To construct the gemcitabine resistant cell lines of human pancreatic cancer cell line (PANC1) and mouse pancreatic cancer cell line (PANC02), and to investigate their biological behavior changes.Methods:Gemcitabine-resistant cell lines PANC1-GR of human pancreatic cancer and PANC02-GR of mouse pancreatic cancer were induced by concentration gradient increment method. Cell count assay (CCK-8), flow cytometry, cell scratch assay and Transwell assay were used to detect the drug resistance, proliferation, cell cycle, migration and invasion of the four groups of cell lines. The drug-resistant cells were also compared with the parent cells.Results:The resistance indices of PANC1-GR and PANC02-GR were 153.3 and 185.4, respectively. The results of CCK-8 showed that with the increase of gemcitabine concentration, the proliferation of resistant cells changed significantly compared with parental cells, the population doubling time of PANC1-GR was significantly shorter than that of PANC1 (1.5±0.1) d vs (2.4±0.2) d ( t=8.00, P<0.001). The proportion of cells in S and G2/M phase increased, and the proportion of cells in G0/G1 phase decreased. The cell scratch and Transwell experiments indicated that the 24h mobility of PANC1-GR and PANC02-GR was higher than that of parent cells (47.6±2.4)% vs (28.7±6.3)% and (53.6±3.2)% vs (30.1±1.4)%, the number of individual field (200 times magnification) penetrating membrane cells was also higher than that of parent cells (269.7±30.9) vs (62.7±10.1) and (172.0±30.8) vs (36.3±4.9), with statistical significance (all P<0.05). Conclusion:Concentration gradient increment method can successfully establish gemcitabine-resistant pancreatic cancer cell lines, which have stronger proliferation, migration and invasiveness, and can be used to study the mechanism of drug resistance in pancreatic cancer.

Objective:To construct the gemcitabine resistant cell lines of human pancreatic cancer cell line (PANC1) and mouse pancreatic cancer cell line (PANC02), and to investigate their biological behavior changes.Methods:Gemcitabine-resistant cell lines PANC1-GR of human pancreatic cancer and PANC02-GR of mouse pancreatic cancer were induced by concentration gradient increment method. Cell count assay (CCK-8), flow cytometry, cell scratch assay and Transwell assay were used to detect the drug resistance, proliferation, cell cycle, migration and invasion of the four groups of cell lines. The drug-resistant cells were also compared with the parent cells.Results:The resistance indices of PANC1-GR and PANC02-GR were 153.3 and 185.4, respectively. The results of CCK-8 showed that with the increase of gemcitabine concentration, the proliferation of resistant cells changed significantly compared with parental cells, the population doubling time of PANC1-GR was significantly shorter than that of PANC1 (1.5±0.1) d vs (2.4±0.2) d ( t=8.00, P<0.001). The proportion of cells in S and G2/M phase increased, and the proportion of cells in G0/G1 phase decreased. The cell scratch and Transwell experiments indicated that the 24h mobility of PANC1-GR and PANC02-GR was higher than that of parent cells (47.6±2.4)% vs (28.7±6.3)% and (53.6±3.2)% vs (30.1±1.4)%, the number of individual field (200 times magnification) penetrating membrane cells was also higher than that of parent cells (269.7±30.9) vs (62.7±10.1) and (172.0±30.8) vs (36.3±4.9), with statistical significance (all P<0.05). Conclusion:Concentration gradient increment method can successfully establish gemcitabine-resistant pancreatic cancer cell lines, which have stronger proliferation, migration and invasiveness, and can be used to study the mechanism of drug resistance in pancreatic cancer.