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With a poor prognosis and lack of effective therapies, pancreatic cancer is one of the most deadly tumors. The early diagnosis of pancreatic cancer is particularly important to improve patients’ survival rates. In the present study, leveraging on the targeted binding of asparagine-glycine-arginine (NGR) peptides to CD13, which is overexpressed on pancreatic cancer cells, we constructed nanoparticles, termed QDs-NGR, and explored the targeted fluorescence imaging and influence on pancreatic cancer at the cellular level in vitro of these nanoparticles. The QDs-NGR nanoparticles enabled the targeted fluorescence imaging of cells, compared with the QDs without the targeting unit, and had obvious inhibiting effects on cancer cells. We have successfully developed a promising nanoparticle with potential for the targeted diagnosis and treatment of pancreatic cancer in vitro, which provides the foundation for further research.