Three new polyhydroxylated spirostanol steroidal saponins, dulongenosides B–D (2–4), along with 14 known compounds, dulongenoside A (1), padelaoside B (5), parisyunnanoside G (6), polyphyllin D (7), ophiopogonin Cʹ (8), formosanin C (9), dioscin (10), paris saponin VII (11), paris H (12), parisyunnanoside I (13), protodioscin (14), proprotogracillin (15), crustecdysone (16), and stigmasterol‐3‐O‐β‐d‐glucopyranoside (17), were isolated from the rhizomes of Paris dulongensis (Melanthiaceae). Their chemical structures were elucidated based on extensive analyses of NMR and MS data and acidic hydrolyses. The isolates were evaluated for their cytotoxicity to five human cancer cell lines (HL‐60, SW480, MDA‐MB‐231, A549, and A549/Taxol) and the normal human bronchial epithelial cell line BEAS‐2B by the MTS test. Compounds 7–12 and 14 showed cytotoxic activity, with IC50 values ranging from 0.20 to 4.35 μM. Proprogracillin selectively inhibited A549(IC50 = 0.58 μM) and A549/Taxol (IC50 = 0.74 μM) cells, with no significant cytotoxic activity against HL‐60, SW480, MDA‐MB‐231, or BEAS‐2B cells, with IC50 values greater than 40 μM.