Thermo/pH-sensitive/fluorescent/biocompatible nanospheres consisting of quantum dots-embedded mesoporous silica nanoparticles (Q-MS) as a core and poly(N-isopropylacrylamide (NIPAM))-graft-chitosan (CS) nanogels as a shell (PNIPAM-g-CS) were prepared via temperature-regulated one-pot copolymerization of NIPAM monomer and CS in the presence of Q-MS. The prepared nanospheres exhibited remarkable fluorescence/thermo/pH-sensitivity. HepG2 cells treated with nanospheres displayed bright fluorescence imaging. Loading efficiency and capacity of Doxorubicin (Dox) into nanospheres were regularly increased with the increment of Dox concentration. At a high temperature and a low pH, cumulative in vitro release of Dox from Dox-loaded nanospheres was much great and fast. Released Dox still retained high anticancer activity, and blank nanosphere carriers produced neglectful toxicity to HepG2 cells. The multifunctional nanospheres could be further developed toward temperature/pH-regulated drug carriers for in vivo tumor therapy with a rapid drug release and fluorescence imaging in targeted tissues.