Objective：To investigate the mechanism of miR-141 regulating cell proliferation in pancreatic cancer. Methods：Expression of miR-141 in normal pancreas cell and four pancreatic cancer cell lines were detected by quantitative reverse transcription PCR（qRT-PCR）. miR-141 expression file of GSE71533 which including 36 pancreatic cancer samples and 16 normal samples was downloaded from Gene Expression Omnibus（GEO）to analyze the expression of miR-141. Western blot was used to detect the expression of Bmi-1 in normal pancreatic cell and four pancreatic cancer cells Luciferase reporter assay was performed to analyze the relationship between Bmi-1 and miR-141. The effects of miR-141 and Bmi-1 on cell proliferation was examined by CCK8 and colony formation assay，respectively. Results：The qRT-PCR results showed that miR-141 was significantly down-regulated in pancreatic cancer cells. miR-141 was down-regulated in pancreatic cancer tissues by analyzing the microarray of GSE71533. Western blot analysis results demonstrated that Bmi-1 was up-regulated in pancreatic cancer cells. The dual luciferase assay indicated that miR-141 was anchored at the 3′-untranslated region of Bmi-1. Down-regulation of Bmi-1 inhibited the proliferation of pancreatic cancer cells. Upregulating miR-141 decreased the protein level of Bmi-1，thus repressing cell proliferation in pancreatic cancer cell. Conclusions：miR-141 acts as a tumor-suppressor in pancreatic cancer by targeting Bmi-1. These findings revealed that miR-141’s potential function in the treatment of pancreatic cancer.