To study the negative bending behavior of the prestressed hollow core slab (HCS) with cast-in-place (CIP) RC layer (HCSPR) in the beam-slab joint area, eight full-scale tests were conducted, covering the following parameters: support length (S), longitudinal reinforcement ratio (ρr), HCS height (h1), and CIP concrete layer height (h2). The mechanical properties of specimens were evaluated including load-deflection relationship, cracking behavior, and strain development. Two distinct failure modes of flexure and shear were observed. An extensive parametric study was also conducted to study the effects of h2, h1, prestressing degree, ρr, and span on negative bending performance. The results from the tests and parametric study indicate that h1, ρr, and h2 significantly affect the failure modes, bearing capacity, and ductility. To ensure robustness, S values no less than 30 mm are recommended for the connection between HCS and RC beam. Moreover, to avoid shear failure of HCS with CIP RC layer (HCSPR) in the joint area under negative moment, the shear span-to-depth ratio [L0/(h1 +h2)] should be restricted to be larger than 4. Lastly, both flexural and shear capacity formulas based on GB50010, EN1168, and ACI318 were compared. The calculated results were compared with the test and parametric analysis results. This research is of great significance for the application of simplified RC beam-HCS connection.