Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway
Flaccidoxide-13-acetate, an active compound isolated from cultured-type soft coral <i>Sinularia gibberosa</i>, has been shown to have inhibitory effects against invasion and cell migration of RT4 and T24 human bladder cancer cells. In our study, we used an 3-(4,5-dimethyl-2-thiazolyl)-2,...
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MDPI AG
2019-05-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/17/5/287 |
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| author | Yu-Jen Wu Tzu-Rong Su Guo-Fong Dai Jui-Hsin Su Chih-I Liu |
| author_facet | Yu-Jen Wu Tzu-Rong Su Guo-Fong Dai Jui-Hsin Su Chih-I Liu |
| author_sort | Yu-Jen Wu |
| collection | DOAJ |
| description | Flaccidoxide-13-acetate, an active compound isolated from cultured-type soft coral <i>Sinularia gibberosa</i>, has been shown to have inhibitory effects against invasion and cell migration of RT4 and T24 human bladder cancer cells. In our study, we used an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), colony formation assay, and flow cytometry to determine the mechanisms of the anti-tumor effect of flaccidoxide-13-acetate. The MTT and colony formation assays showed that the cytotoxic effect of flaccidoxide-13-acetate on T24 and RT4 cells was dose-dependent, and the number of colonies formed in the culture was reduced with increasing flaccidoxide-13-acetate concentration. Flow cytometry analysis revealed that flaccidoxide-13-acetate induced late apoptotic events in both cell lines. Additionally, we found that flaccidoxide-13-acetate treatment upregulated the expressions of cleaved caspase 3, cleaved caspase 9, Bax, and Bad, and down-regulated the expressions of Bcl-2, <i>p</i>-Bad, Bcl-x1, and Mcl-1. The results indicated that apoptotic events were mediated by mitochondrial dysfunction via the caspase-dependent pathway. Flaccidoxide-13-acetate also provoked endoplasmic reticulum (ER) stress and led to activation of the PERK-eIF2<i>α</i>-ATF6-CHOP pathway. Moreover, we examined the PI3K/AKT signal pathway, and found that the expressions of phosphorylated PI3K (<i>p</i>-PI3K) and AKT (<i>p</i>-AKT) were decreased with flaccidoxide-13-acetate concentrations. On the other hand, our results showed that the phosphorylated JNK and p38 were obviously activated. The results support the idea that flaccidoxide-13-acetate-induced apoptosis is mediated by mitochondrial dysfunction, ER stress, and activation of both the p38 and JNK pathways, and also relies on inhibition of PI3K/AKT signaling. These findings imply that flaccidoxide-13-acetate has potential in the development of chemotherapeutic agents for human bladder cancer. |
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| language | English |
| last_indexed | 2024-04-13T06:42:04Z |
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| spelling | doaj.art-608e39b80a614174aff0f0cb2f90348d2022-12-22T02:57:43ZengMDPI AGMarine Drugs1660-33972019-05-0117528710.3390/md17050287md17050287Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated PathwayYu-Jen Wu0Tzu-Rong Su1Guo-Fong Dai2Jui-Hsin Su3Chih-I Liu4Department of Nursing, Meiho University, Pingtung 91202, TaiwanAntai Medical Care Cooperation Antai Tian-Sheng Memorial Hospital, Pingtung 92842, TaiwanDepartment of Biological Technology, Meiho University, Pingtung 91202, TaiwanNational Museum of Marine Biology and Aquarium, Pingtung 94450, TaiwanDepartment of Nursing, Meiho University, Pingtung 91202, TaiwanFlaccidoxide-13-acetate, an active compound isolated from cultured-type soft coral <i>Sinularia gibberosa</i>, has been shown to have inhibitory effects against invasion and cell migration of RT4 and T24 human bladder cancer cells. In our study, we used an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), colony formation assay, and flow cytometry to determine the mechanisms of the anti-tumor effect of flaccidoxide-13-acetate. The MTT and colony formation assays showed that the cytotoxic effect of flaccidoxide-13-acetate on T24 and RT4 cells was dose-dependent, and the number of colonies formed in the culture was reduced with increasing flaccidoxide-13-acetate concentration. Flow cytometry analysis revealed that flaccidoxide-13-acetate induced late apoptotic events in both cell lines. Additionally, we found that flaccidoxide-13-acetate treatment upregulated the expressions of cleaved caspase 3, cleaved caspase 9, Bax, and Bad, and down-regulated the expressions of Bcl-2, <i>p</i>-Bad, Bcl-x1, and Mcl-1. The results indicated that apoptotic events were mediated by mitochondrial dysfunction via the caspase-dependent pathway. Flaccidoxide-13-acetate also provoked endoplasmic reticulum (ER) stress and led to activation of the PERK-eIF2<i>α</i>-ATF6-CHOP pathway. Moreover, we examined the PI3K/AKT signal pathway, and found that the expressions of phosphorylated PI3K (<i>p</i>-PI3K) and AKT (<i>p</i>-AKT) were decreased with flaccidoxide-13-acetate concentrations. On the other hand, our results showed that the phosphorylated JNK and p38 were obviously activated. The results support the idea that flaccidoxide-13-acetate-induced apoptosis is mediated by mitochondrial dysfunction, ER stress, and activation of both the p38 and JNK pathways, and also relies on inhibition of PI3K/AKT signaling. These findings imply that flaccidoxide-13-acetate has potential in the development of chemotherapeutic agents for human bladder cancer.https://www.mdpi.com/1660-3397/17/5/287flaccidoxide-13-acetatecultured soft coral <i>Sinularia gibberosa</i>human bladder cancerapoptosismitochondrial dysfunctionPERK-eIF2<i>α</i>-ATF6-CHOP pathwayp38/JNK |
| spellingShingle | Yu-Jen Wu Tzu-Rong Su Guo-Fong Dai Jui-Hsin Su Chih-I Liu Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway Marine Drugs flaccidoxide-13-acetate cultured soft coral <i>Sinularia gibberosa</i> human bladder cancer apoptosis mitochondrial dysfunction PERK-eIF2<i>α</i>-ATF6-CHOP pathway p38/JNK |
| title | Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway |
| title_full | Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway |
| title_fullStr | Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway |
| title_full_unstemmed | Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway |
| title_short | Flaccidoxide-13-Acetate-Induced Apoptosis in Human Bladder Cancer Cells is through Activation of p38/JNK, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress Regulated Pathway |
| title_sort | flaccidoxide 13 acetate induced apoptosis in human bladder cancer cells is through activation of p38 jnk mitochondrial dysfunction and endoplasmic reticulum stress regulated pathway |
| topic | flaccidoxide-13-acetate cultured soft coral <i>Sinularia gibberosa</i> human bladder cancer apoptosis mitochondrial dysfunction PERK-eIF2<i>α</i>-ATF6-CHOP pathway p38/JNK |
| url | https://www.mdpi.com/1660-3397/17/5/287 |
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