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Background: Pancreatic Ductal Adenocarcinoma (PDAC) is the most common form of pancreatic cancer and leading causes of pancreatic cancer death because of most PDAC patients with advanced unresectable disease at that time, which is remarkably resistant to all forms of chemotherapy and radiotherapy. Objective: PDAC increases the social and patients family burden. However, the PDAC pathogenesis is not identified. We are trying to uncover the underlying mechanism in the future. Methods: In our research, drug-resistant cell line was successfully induced in the vitro by progressive concentrations of Afatinib, which we named it as BxPC3-AR. Results: We found that the effect of autophagy was on the resistance of BxPC3-AR to Afatinib. Conclusion: In a word, it is confirmed that the autophagy plays a certain role in BxPC3-AR resistance to Afatinib. Our findings provide new perspective into the role of autophagy in pancreatic ductal adenocarcinoma.