A novel nanoconjugate enabled synchronization of localized hyperthermia and drug release was presented. The hyperthermia was induced by the radiofrequency (RF) at 1.3MHz to trigger the release of 5-fluouracil (5-FU) anti-cancer drugs from magnetite nanocrystals core by breaking their hydrogen bond with the polynucleotide polymer carrier. The anti-Human Epidermal Growth Factor Receptor 2 (anti-HER2) antibody was end-modified on the polyethylene glycol to achieve cancer cell specific targeting and biological stealth. The 22 nm-sized magnetites displayed high saturation magnetization (94 emu/g) comparable to bulk magnetite (92 emu/g). The local regional heat triggers synchronized drug release and physical thermal damage of the cancer cells and significantly improved the anti-cancer efficacy in both in vitro and in vivo models than hyperthermia or chemotherapy alone as evidenced by cell viability assays and pathological examines. Complete remission of the disease in vivo was achieved through radiofrequency synchronization of hyperthermia and chemotherapy after intravenous injection of the nanoconjugates.