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Objective To explore the influences of SLC39A5 knockout on the establishment of esophageal cancer model induced by 4-nitroquinoline 1-oxide (4-NQO) in C57BL/6 mice. Methods Ten wild-type mice were treated as negative control group and drank the 1, 2-propylene glycol at 100 μg/ml; 140 wild-type mice and 80 knockout genotype mice were treated as experimental groups and drank the carcinogen 4-NQO stock solution dissolved in 1, 2-propylene glycol at 100μg/ml, the experimental time was 28 weeks and all three groups' mice were sacrificed. Results The survival rates were 100%, 92.96% and 91.25% in negative group, wild-type experimental group and SLC39A5 knockout genotype experimental group, respectively; the rates of tumor formation were 0, 61.36% and 28.77%, and there was a statistical difference between the two experimental groups(χ2=19.98, P < 0.001). Conclusion The esophageal cancer model in C57BL/6 mice and SLC39A5 knockout mice are established successfully and the facilitated role of SLC39A5 in the occurrence and development of esophageal cancer is verified.