In vitro Anticancer Effects of JI017 on Two Prostate Cancer Cell Lines Involve Endoplasmic Reticulum Stress Mediated by Elevated Levels of Reactive Oxygen Species

Abstract

Prostate cancer is the second most commonly diagnosed cancer, and prostate cancer is the second most common cause of cancer death in United States men after lung cancer. Many therapies are used to treat prostate cancer, and chemotherapy is one of the most relevant treatments. However, chemotherapy has many side effects, and repeated administration of chemotherapeutic agents leads to acquired resistance. Thus, new drugs with few side effects are needed. We investigated the molecular mechanism of action of JI017 in human prostate cancer cells. We identified an endoplasmic reticulum (ER) stress pathway that depended on the reactive oxygen species (ROS) pathway and played a crucial role in JI017-induced apoptosis. We measured cell viability by the MTS assay to determine the effect of JI017. Analysis of apoptosis, mitochondrial dysfunction, and cell cycle features was performed by flow cytometry. We used western blot and RT-PCR to measure the levels of the proteins of the unfolded protein response (UPR) pathway and apoptosis markers. Immunoprecipitation assay and transfection were used to determine the expression levels of proteins interacting with the pathways influenced by JI017 in prostate cancer cells. The anticancer effects induced by JI017 were evaluated. JI017 induced cell death that regulated apoptotic molecules and caused cell cycle arrest that inhibited the proliferation of cancer cells. Moreover, JI017 generated ROS. Accumulation of ROS caused ER stress through the PERK–eIF2α–CHOP and IRE1α-CHOP pathways. Furthermore, persistent activation of the UPR pathway induced by JI017 treatment triggered mitochondrial dysfunction, including dissipation of mitochondrial membrane potential, which activated intrinsic apoptotic pathway in human prostate cancer cells. The data indicated that N-acetyl-L-cysteine diminished apoptosis. We demonstrated that JI017 induced ER stress and cell death. Anticancer properties of JI017 in prostate cancer cells and in a human prostate cancer model involved ROS-mediated ER stress. Thus, JI017 treatment provides a new strategy for chemotherapy of prostate cancer.

JI017 Attenuates Oxaliplatin-Induced Cold Allodynia via Spinal TRPV1 and Astrocytes Inhibition in Mice

“…As Zingiber officinale only constitutes a quarter of JI017, in this study, it might have been in insufficient quantity to be efficacious against both cold and mechanical allodynia. Furthermore, HPLC data identified decursin, nodakenin, aconitine, and 6-gingerol as major phytochemicals of JI017 [37]. Decursin is a major phytochemical of Angelica gigas, while aconitine is present in Aconitum carmichaelii.…”

“…JI017 consists of three medicinal herbs: Aconitum carmichaelii, Angelica gigas, and Zingiber officinale. In a previous study, JI017 showed anti-tumor potency in prostate cancer cell lines [37]. Although the analgesic efficacy of JI017 has never been reported, that of each of its components, in various pain models, has been documented [36,39,41].…”

“…The extract was filtered twice through Whatman grade 2 filter paper (GE Healthcare Life Sciences, Marlborough, MA, USA) to remove insoluble materials. The filtered extract was lyophilized to a powder using a freeze dryer (iLShin biobase, Gyeonggi, South Korea) and stored at 4 • C [37]. For administration, it was dissolved in phosphate-buffered saline (PBS), and the solution was administered orally at a dose of 500 mg/kg.…”

“…JI017 is composed of three medicinal herbs, namely, Aconitum carmichaelii, Angelica gigas, and Zingiber officinale. A recent study has revealed that JI017 has anti-tumor effects on prostate cancer cell lines [37]. Aconitum carmichaelii has been reported to be potent against nerve injury-, oxaliplatin-, and paclitaxel-induced neuropathy [38][39][40].…”

Mitochondria‐associated endoplasmic reticulum membrane: Overview and inextricable link with cancer

“…JI017 (an herb mixture composed of Aconitum carmichaelii, Angelica gigas and Zingiber officinale) exerts anticancer effect in prostate cancer cells via ROS-mediated ER stress. 123 Briefly, JI017 generates ROS and induces ER stress. Stress conditions induce the dissociation of GRP78/BIP (sensor key regulatory protein) from the IRE1α and PERK sensors, and subsequently activate sensor proteins, leading to eIF2α activation.…”