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Involvement of endoplasmic reticulum stress in isoliquiritigenin-induced SKOV-3 cell apoptosis
Xuan Yuan1, Bacui Yu, Yanming Wang
1School of Pharmacy, Shihezi University, 832002, Shihezi, P.R. China.
Abstract:
Isoliquiritigenin (ISL), a licorice chalconoid, is a bioactive agent with chemopreventive potential that has been patented for tumor treatment in China. This study investigated the mechanisms of ISL-induced apoptosis in ovarian carcinoma SKOV-3 cells. Cell viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. The apoptotic rate was determined via flow cytometry using an annexin V-FITC apoptosis detection kit. The intracellular reactive oxygen species (ROS) levels were assessed using a 2,7-dichlorofluorescein probe assay. Malondialdehyde (MDA) formation was determined via thiobarbituric acid reactive substance test. The expressions of growth arrest and DNA damage-inducible gene (GADD153/CHOP), 78 kDa glucose-regulated protein (GRP 78), α-subunit of eukaryotic initiation factor 2 (eIF2α) phosphorylation, activating transcription factor 6α (ATF6α), and unspliced form of X-box binding protein1 (XBP1U) were analyzed via Western blot. Caspase-3 and caspase-12 activities were assessed using a fluorometric kit. Findings indicate that ISL significantly inhibits SKOV-3 cell proliferation, increases intracellular ROS levels, and causes SKOV-3 cell apoptosis. Moreover, ISL-exposed SKOV-3 cells trigger endoplasmic reticulum (ER) stress, as indicated by the enhancement of ER stress-related molecules p-eIF2α, GADD153/CHOP, GRP78, XBP1 expression, and cleavage of ATF6α. However, caspase-12 inhibitor (Z-ATAD) effectively and partially prevents ROS and MDA formation and inhibits ISL-induced SKOV-3 cell apoptosis. ISL induces apoptosis via ER stress-triggered signaling pathways in SKOV-3 cells. ER stress-induced cancer cell apoptosis has been discussed in some patents.
Insights
Isoliquiritigenin (ISL) triggers apoptosis in ovarian cancer cells by inducing reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress. This natural compound shows potential for cancer treatment by activating cell death pathways.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Isoliquiritigenin (ISL), a chalconoid from licorice, exhibits chemopreventive properties and is patented for tumor treatment in China.
- Ovarian carcinoma SKOV-3 cells are a relevant model for investigating anti-cancer mechanisms.
- Understanding the molecular pathways of apoptosis induction is crucial for developing novel cancer therapies.
Purpose of the Study:
- To elucidate the mechanisms by which Isoliquiritigenin (ISL) induces apoptosis in ovarian carcinoma SKOV-3 cells.
- To investigate the role of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress in ISL-mediated apoptosis.
- To identify key molecular players involved in the signaling pathways triggered by ISL.
Main Methods:
- Cell viability assessed using MTT assay.
- Apoptotic rate determined by flow cytometry with Annexin V-FITC.
- Intracellular ROS, malondialdehyde (MDA), and caspase activities measured using specific assays.
- Western blot analysis performed for ER stress markers (GADD153/CHOP, GRP78, p-eIF2α, ATF6α, XBP1U).
Main Results:
- ISL significantly inhibited SKOV-3 cell proliferation and induced apoptosis.
- ISL treatment increased intracellular ROS and malondialdehyde (MDA) levels.
- ISL triggered endoplasmic reticulum (ER) stress, evidenced by elevated p-eIF2α, GADD153/CHOP, GRP78, and XBP1 expression, and ATF6α cleavage.
- Caspase-12 inhibition partially blocked ROS/MDA formation and ISL-induced apoptosis.
Conclusions:
- Isoliquiritigenin (ISL) induces apoptosis in ovarian carcinoma SKOV-3 cells.
- ISL-induced apoptosis is mediated through the activation of ER stress-signaling pathways.
- The findings support the potential of ISL as a therapeutic agent for ovarian cancer, leveraging ER stress-induced cell death mechanisms.