Osthole induces apoptosis of the HT‑29 cells via endoplasmic reticulum stress and autophagy

Abstract

Endoplasmic reticulum stress (ERS) and autophagy are important pathways, which induce apoptosis of tumor cells. Osthole has been demonstrated to exert anticancer effects via the induction of apoptosis in several human colon cancer lines, but the mechanism underlying its involvement in the induction of ERS and autophagy in the human HT-29 colorectal cancer cell line remains unknown. The present study aimed to identify the possible signaling pathways involved in osthole-induced apoptosis of HT29 cells. Methodologically, colony formation and Cell Counting Kit-8 assays were used to assess cell proliferation and viability, respectively, while flow cytometry was performed to investigate apoptosis. Signaling pathways, including apoptosis, autophagy and ERS, were also investigated in the HT-29 cell line using western blot analysis. The results demonstrated that osthole inhibited cellular proliferation and viability in a dose-dependent manner. In addition, osthole induced the expression level of proteins associated with mitochondria-mediated cell apoptosis, autophagy and ERS. The association between autophagy and ERS in osthole-induced apoptosis in the HT-29 cell line was further clarified. Inhibiting cell autophagy with the inhibitor, 3-methyladenine, suppressed osthole-induced cell apoptosis and enhanced osthole-induced ERS. By contrast, alleviating ERS with the inhibitor, 4-phenylbutyric acid attenuated osthole-induced cell apoptosis and autophagy. In conclusion, osthole could significantly suppress the proliferation and viability of the HT-29 colorectal cancer cell line and induce cell apoptosis via autophagy and ERS. Furthermore, ERS may play a more important role in osthole-induced cell apoptosis.

Potential roles and molecular mechanisms of phytochemicals against cancer

“…81 Osthole suppresses cell proliferation and viability, and induces cell apoptosis by activating autophagy and the ER stress pathway in HT-29 cells. 82 Centipeda minima , a traditional Chinese herbal medicine, has been demonstrated to handle rhinitis and relieve pain in humans. It could hinder diverse pathways, including the EGFR, PI3K/AKT/mTOR, NF-κB, and STAT3 pathways to inhibit cell proliferation and MMP-9-mediated cell migration, and induce cell apoptosis in triple-negative breast cancer (TNBC) cells and in vivo .…”

Osthole: An up-to-date review of its anticancer potential and mechanisms of action

“…Furthermore, cotreatment of hypoxic HCT116 cells with osthole greatly increased the sensitivity to cisplatin ( Peng and Chou, 2022 ). In another study, osthole (100, 50 and 25 µM) could significantly suppress cellular proliferation and viability in a colorectal cancer cell line (HT-29) treated with 0.1% DMSO as the control and induce cell apoptosis via autophagy and endoplasmic reticulum stress ( Zhou et al, 2021 ).…”

Traditional Chinese medicine suppressed cancer progression by targeting endoplasmic reticulum stress responses: A review

“…In HT-29 colon cancer cells, the ERS signaling proteins GRP78, p-PERK/PERK, p-elF2α/elF2α, and CHOP were upregulated and apoptosis was increased after serpentine treatment. [ 38 ] Inhibition of cellular autophagy using 3-methyladenine suppresses apoptosis induced by serpentine via the ERS pathway. Mitigation of ERS with a 4-phenylbutyric acid inhibitor also attenuates serpentine-induced apoptosis and autophagy.…”

“…Head and neck cancer 1 Glaucocalyxin A ATF4/ CHOP/CHAC1 pathways [9] 2 Tetrandrine GRP78 and GADD153 [10] Digestive tract tumor 3 Tanshinone IIA BIP and CHOP [11] 4 Wogonoside IRE1α-TRAF2-ASK1 pathways [12] 5 Cyperus amuricus GRP78/BiP; CHOP-URP-PERK/IRE1/ATF6 [13] 6 Muscone PERK/ATF4/DDIT3 pathways [14] 7 Celastrol IRE1α-XBP1s pathways; GRP78/BiP, ATF4 and CHOP [15] 8 Salidroside p-PERK, p-eIF2a, ATF-6 and CHOP [16] 9 TTF1-NPs GRP78, PERK, IRE1α, ATF6 and CHOP; PERK-eIF2α-ATF4 and ATF6 [17,18] 10 Epigallocatechin-3-gallate p-PERK and p-eIF2α [19] 11 Shikonin PERK/elF2α/ATF4/CHOP, and IRE1α/JNK [20] 12 Gambogenic acid IRE1α, BiP, p-eIF2α, and ATF4 [21] 13 A purified resin glycoside fraction from pharbitidis semen ERS-JNK pathways [22] Gynecologic urogenital tumors 14 Saikosaponin-A GPR78 and CHOP [23] 15 Identification of dihydrotanshinone I PERK [24] Non-small cell lung cancer 16 Kushenol Z CHOP [25] 17 Evodiamine IRE1α-TRAF2-ASK1-JNK [26] 18 flavonoid components in Astragali Radix IRE1α-XBP1s [27] Others 19 Shikonin p-eIF2α and CHOP [28] 20 Psoralen CHOP, IREa, XBP-1s, GRP78, PERK and ATF-6 [29] 21 Artesunate ATF4-CHOP-CHAC1 [30] 22 (-)-Epigallocatechin-3-gallate GRP78, ATF4, XBP1s, CHOP and EDEM [31] 23 Asperuloside GRP78, p-PERK, p-eIF2α, CHOP, p-IRE1, XBP1, and ATF6 [32] Tumor cell autophagy Non-small cell lung cancer 24 Ginsenoside ATF4-CHOP-AKT1-mTOR [36] 25 Guangsangon E (GSE) ROS-ERS (GRP78, IRE1α and ATF4) [37] Colorectal cancer 26 Serpentine GRP78, p -PERK, p-elF2α and CHOP [38] 27 Cryptotanshinone Cell autophagy by ERS [39] Gynecologic urogenital tumors 28 Cucurbitacin-I IRE1α-caspase 1-p-PERK-eIF2-ATF4-CHOP [40] Tumor cell invasion and metastasis…”