To examine whether the combination of the TC-HT and CGA- or EGCG-induced apoptosis involved mitochondrial disruption, the mitochondrial membrane potential (MMP) was assessed using DiOC6(3) fluorescence staining by flow cytometric analysis . GUID:?6D1664B1-3F6E-4728-81C3-7A198AA913B8 Data Availability StatementAll relevant data are within the paper and its Supporting Information files. Abstract Hyperthermia (HT) has shown feasibility and potency as an anticancer therapy. Administration of HT in the chemotherapy has previously enhanced the cytotoxicity of drugs against pancreatic cancer. However, the drugs used when conducting these studies are substantially conventional chemotherapeutic agents that may cause unwanted side effects. Additionally, the thermal dosage in the treatment of cancer cells could also probably harm the healthy cells. The purpose of this work was to investigate the potential of the two natural polyphenolic compounds, epigallocatechin gallate (EGCG) and chlorogenic acid (CGA), as heat synergizers in the thermal treatment of the PANC-1 cells. CGK 733 Furthermore, we have introduced a unique strategy entitled the thermal cycling-hyperthermia (TC-HT) that is capable of providing a maximum synergy and minimal side effect with the anticancer compounds. Our results demonstrate that the combination of the TC-HT and the CGA or EGCG markedly exerts the anticancer effect against the PANC-1 cells, while none of the single treatment induced such changes. The synergistic activity was attributed to the cell cycle arrest at the G2/M phase and the induction of the ROS-dependent mitochondria-mediated apoptosis. These findings not only represent the first thermal synergistic study of CGK 733 natural compounds in the treatment of pancreatic cancer, but also highlight the potential of the TC-HT as an alternative strategy in thermal treatment. Introduction Pancreatic cancer is one of the leading causes in cancer death and remains one of the deadliest solid human malignancies worldwide . Patients with pancreatic cancer are commonly diagnosed at the unresectable stage, and in most cases, patients with advanced pancreatic cancer have a poor response to chemotherapy or radiotherapy. In spite of the fact that therapeutic methods have been improved, the prognosis for pancreatic cancer patients still remains poor with a low five-year survival rate . Therefore, there is a need for continued CGK 733 research in novel agents or alternative therapeutic strategies for treating pancreatic cancers, thereby making an improvement for the patients quality of life. Hyperthermia (HT) has emerged as a promising method for treating cancer over the past decades . It is a procedure exposing the tumor tissue to high temperatures that cause cancer cell damage and death. Researches have shown that HT exhibits therapeutic potential against cancer cells through multiple cellular changes, such as protein denaturation and aggregation, inhibition of DNA synthesis, cytoskeleton disruption, and alteration in the calcium homeostasis [4C6]. In addition, HT can HSF directly activate the immune response against the tumors, increase the tumor oxygenation, and improve the drug delivery [7C9]. Although these encouraging results have expanded our understanding of the cytotoxic effects of HT on the cancer cells, in the case of HT as single treatment, it has been shown not to be sufficient to kill cancer cells . To strengthen the effectiveness of HT, several investigations have explored combinations of HT and other cancer therapies, such as radiotherapy and chemotherapy . It has been demonstrated to be effective against various types of cancer, including pancreatic cancer, in that HT enhanced the cytotoxicity of gemcitabine through the inhibition of nuclear factor kappa B (NF-B) [12C14]. There have also been reports of gemcitabine and other drugs, such as cisplatin and carbonplatin, combined with HT, that demonstrated the clinical efficacy in patients with pancreatic cancer [15, 16]. These data suggest that HT could modify the cytotoxicity of the anticancer drugs, thereby yielding better outcomes in treating pancreatic cancer. However, the drugs used in these combined treatments are conventional chemotherapeutic drugs, which have been known to cause unpleasant and even dangerous side effects. Nowadays, there has been an increasing interest in natural compounds research due to their lower toxicity and diverse CGK 733 biological properties. Phenolic compounds are among the most studied in cancer prevention and cure, and also the largest group of phytochemicals, as well as being widely distributed in our diet. Particularly, regular intakes of dietary polyphenols have been linked to lower risks of many cancers . Tea and coffee are two of the most consumed beverages worldwide, and the natural phenolic compounds, epigallocatechin gallate (EGCG) and chlorogenic acid (CGA), are.