Abstract: Objective 　Comparison of the effects of pirarubicin (THP) in combination with hyperthermia on gastric cancer tissues in vitro and to discuss the underlying mechanisms. Methods 　In vitro three-dimensional acting models were established with tissue biopsies from 36 patients with pathologically confirmed gastric cancer. Tumor cell viability was measured by MTS2PMS assay. TUNEL fluorescence detection in situ apoptosis, HE staining was used to study histomorphology of drug treated gastric tissues. Electron microscope observed cells. Results 　Synergistic effects were observed after tumor tissues were treated with hyperthermia and three drugs including cisplatin, THP, and mitomycin ( P= 0. 000), while THP exhibited superior cytotoxic effects to other drugs on cancer tissues. Furthermore, histomorphological studies suggested strong killing effects of THP on cancer tissues, including disrupted tissue structure, cellular degradation and necrosis, karyopyknosis and karyolysis, also with loss of cytoplasm. Antitumoral effects of THP were associated with pathological grading ( P = 0. 000), while irrelated to patients' gender, age, tumor size, clinical staging and preoperative CEA levels( P > 0. 05) . The apoptosis index has significant difference compared THP and the other three drugs. After treated by THP, the gastric tissues disaggregation, and electron microscope observed cell membrane invagination, endochylema vacuolization. Nuclear membrane diaappeared. Conclusion 　Synergistic effects of pirarubicin in combination with hyperthermia were evident ; cytotoxicity of pirarubicin was thermally enhanced considerably by mild hyperthermia on gastric cancer tissue. THP may be a potential therapeutic drug for intraperitoneal chemohyperthermia.