Diallyl Disulfide Induces Apoptosis and Autophagy in Human Osteosarcoma MG-63 Cells through the PI3K/Akt/mTOR Pathway

Diallyl disulfide (DADs), a natural organic compound, is extracted from garlic and scallion and has anti-tumor effects against various tumors. This study investigated the anti-tumor activity of DADs in human osteosarcoma cells and the mechanisms. MG-63 cells were exposed to DADs (0, 20, 40, 60, 80, and 100 μM) for different lengths of time (24, 48, and 72 h). The CCK8 assay results showed that DADs inhibited osteosarcoma cell viability in a dose-and time-dependent manner. FITC-Annexin V/propidium iodide staining and flow cytometry demonstrated that the apoptotic ratio increased and the cell cycle was arrested at the G2/M phase as the DADs concentration was increased. A Western blot analysis was employed to detect the levels of caspase-3, Bax, Bcl-2, LC3-II/LC3-I, and p62 as well as suppression of the mTOR pathway. High expression of LC3-II protein revealed that DADs induced formation of autophagosome. Furthermore, DADs-induced apoptosis was weakened after adding 3-methyladenine, demonstrating that the DADs treatment resulted in autophagy-mediated death of MG-63 cells. In addition, DADs depressed p-mTOR kinase activity, and the inhibited PI3K/Akt/mTOR pathway increased DADs-induced apoptosis and autophagy. In conclusion, our results reveal that DADs induced G2/M arrest, apoptosis, and autophagic death of human osteosarcoma cells by inhibiting the PI3K/Akt/mTOR signaling pathway.

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Antiproliferative Properties of Oleuropein in Human Osteosarcoma Cells

Natural Product Communications ◽

10.1177/1934578x1601100418 ◽

2016 ◽

Vol 11 (4) ◽

pp. 1934578X1601100 ◽

Cited By ~ 1

Author(s):

Jose M. Moran ◽

Olga Leal-Hernandez ◽

Maria L. Canal-Macías ◽

Raul Roncero-Martin ◽

Rafael Guerrero-Bonmatty ◽

...

Keyword(s):

Cell Lines ◽

Osteosarcoma Cell ◽

Dependent Manner ◽

Osteosarcoma Cells ◽

Probit Regression ◽

Cytotoxic Effects ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Mg63 Cells ◽

Human Osteosarcoma Cell

In this study, we evaluated the antiproliferative activity on two human osteosarcoma cell lines (MG-63 and Saos2) of oleuropein, an olive oil compound traditionally found in the Mediterranean diet. Oleuropein exhibited obvious cytotoxic effects on human osteosarcoma cells in a concentration- and time-dependent manner. Statistical analysis of IC50 by the Probit regression method suggested that oleuropein had similar toxic effects on both cell lines tested (IC50 range from 247.4–475.0 μM for MG63 cells and from 798.7–359.9 μM for Saos2 cells).

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Neohesperidin Induces Cell Cycle Arrest, Apoptosis, and Autophagy via the ROS/JNK Signaling Pathway in Human Osteosarcoma Cells

The American Journal of Chinese Medicine ◽

10.1142/s0192415x21500609 ◽

2021 ◽

pp. 1-24

Author(s):

Shubin Wang ◽

Zongguang Li ◽

Wei Liu ◽

Guojun Wei ◽

Naichun Yu ◽

...

Keyword(s):

Cell Cycle ◽

Flow Cytometry ◽

Signaling Pathway ◽

Osteosarcoma Cell ◽

Osteosarcoma Cells ◽

Jnk Signaling ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Jnk Signaling Pathway ◽

Induced Apoptosis

Neohesperidin has anti-oxidative and anti-inflammatory properties and exerts extensive therapeutic effects on various cancers. In this study, the osteosarcoma cell lines were exposed to different concentrations of neohesperidin. Cell proliferation and viability were assessed by CCK-8 and colony-formation assays. The role of neohesperidin in cell cycle progression and apoptosis were analyzed by flow cytometry and western blotting. To identify autophagosomes and autolysosomes, we used a tandem GFP-mRFP-LC3B lentiviral construct. In addition, autophagy was evaluated by examining autophagosome formation using transmission electron microscopy. Intracellular reactive oxygen species (ROS) production was detected by fluorescence microscopy and flow cytometry. Subsequently, the activation of the ROS/JNK signaling pathway was investigated. Neohesperidin could inhibit proliferation and induce apoptosis in SJSA and HOS cells. The formation of autophagosomes indicated that autophagy occurred in neohesperidin-treated cells and the apoptotic effect of neohesperidin was significantly increased after the use of autophagy inhibitors. Subsequently, we found that neohesperidin-induced apoptosis and autophagy were related to the increase in ROS generation and were significantly inhibited by GSH. Moreover, neohesperidin induced activation of the c-Jun N-terminal kinase (JNK) signaling pathway and inhibition of JNK with SP600125 attenuated neohesperidin-induced apoptosis and autophagy simultaneously. Our data indicated that neohesperidin caused G2/M phase arrest and induced apoptosis and autophagy by activating the ROS/JNK pathway in human osteosarcoma cells, suggesting that neohesperidin is a potential drug candidate for the treatment of osteosarcomas.

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Andrographolide Induces Autophagic Cell Death and Inhibits Invasion and Metastasis of Human Osteosarcoma Cells in An Autophagy-Dependent Manner

Cellular Physiology and Biochemistry ◽

10.1159/000485536 ◽

2017 ◽

Vol 44 (4) ◽

pp. 1396-1410 ◽

Cited By ~ 9

Author(s):

Ying Liu ◽

Yan Zhang ◽

Jilong Zou ◽

Lixin Yan ◽

Xiufeng Yu ◽

...

Keyword(s):

Cell Death ◽

Morphological Changes ◽

Annexin V ◽

Beclin 1 ◽

Migration And Invasion ◽

Dependent Manner ◽

Osteosarcoma Cells ◽

Invasion And Metastasis ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells

Background/Aims: Osteosarcoma (OS) is the most common primary malignant tumor of bone tissue. Although treatment effectiveness has improved, the OS survival rate has fluctuated in recent years. Andrographolide (AG) has been reported to have antitumor activity against a variety of tumors. Our aim was to investigate the effects and potential mechanisms of AG in human osteosarcoma. Methods: Cell viability and morphological changes were assessed by MTT and live/dead assays. Apoptosis was detected using Annexin V-FITC/PI double staining, DAPI, and caspase-3 assays. Autophagy was detected with mRFP-GFP-LC3 adenovirus transfection and western blot. Cell migration and invasion were detected by wound healing assay and Transwell® experiments. Results: AG dose-dependently reduced the viability of osteosarcoma cells. No increase in apoptosis was detected in AG-treated human OS MG-63 and U-2OS cells, and the pan-caspase inhibitor z-VAD did not attenuate AG-induced cell death. However, AG induced autophagy by suppressing PI3K/Akt/mTOR and enhancing JNK signaling pathways. 3-MA and Beclin-1 siRNA could reverse the cytotoxic effects of AG. In addition, AG inhibited the invasion and metastasis of OS, and this effect could be reversed with Beclin-1 siRNA. Conclusion: AG inhibits viability and induces autophagic death in OS cells. AG-induced autophagy inhibits the invasion and metastasis of OS.

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CLEFMA Activates the Extrinsic and Intrinsic Apoptotic Processes through JNK1/2 and p38 Pathways in Human Osteosarcoma Cells

Molecules ◽

10.3390/molecules24183280 ◽

2019 ◽

Vol 24 (18) ◽

pp. 3280 ◽

Cited By ~ 2

Author(s):

Jia-Sin Yang ◽

Renn-Chia Lin ◽

Yi-Hsien Hsieh ◽

Heng-Hsiung Wu ◽

Geng-Chung Li ◽

...

Keyword(s):

Annexin V ◽

Synthetic Analog ◽

Osteosarcoma Cells ◽

Apoptotic Pathway ◽

Term Survival ◽

Anticancer Properties ◽

Human Osteosarcoma ◽

Effector Caspase ◽

Human Osteosarcoma Cells ◽

Induced Apoptosis

Due to the poor prognosis of metastatic osteosarcoma, chemotherapy is usually employed in the adjuvant situation to improve the prognosis and the chances of long-term survival. 4-[3,5-Bis(2-chlorobenzylidene)-4-oxo-piperidine-1-yl]-4-oxo-2-butenoic acid (CLEFMA) is a synthetic analog of curcumin and possesses anti-inflammatory and anticancer properties. To further obtain information regarding the apoptotic pathway induced by CLEFMA in osteosarcoma cells, microculture tetrazolium assay, annexin V-FITC/PI apoptosis staining assay, human apoptosis array, and Western blotting were employed. CLEFMA dose-dependently decreased the cell viabilities of human osteosarcoma U2OS and HOS cells and significantly induced apoptosis in human osteosarcoma cells. In addition to the effector caspase 3, CLEFMA significantly activated both extrinsic caspase 8 and intrinsic caspase 9 initiators. Moreover, CLEFMA increased the phosphorylation of extracellular signal-regulated protein kinases (ERK)1/2, c-Jun N-terminal kinases (JNK)1/2 and p38. Using inhibitors of JNK (JNK-in-8) and p38 (SB203580), CLEFMA’s increases of cleaved caspases 3, 8, and 9 could be expectedly suppressed, but they could not be affected by co-treatment with the ERK inhibitor (U0126). Conclusively, CLEFMA activates both extrinsic and intrinsic apoptotic pathways in human osteosarcoma cells through JNK and p38 signaling. These findings contribute to a better understanding of the mechanisms responsible for CLEFMA’s apoptotic effects on human osteosarcoma cells.

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Ionizing radiation enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis through up-regulations of death receptor 4 (DR4) and death receptor 5 (DR5) in human osteosarcoma cells

Journal of Orthopaedic Research® ◽

10.1002/jor.21056 ◽

2009 ◽

Vol 28 (6) ◽

pp. 739-745 ◽

Cited By ~ 20

Author(s):

Takeshi Hori ◽

Takashi Kondo ◽

Masahiko Kanamori ◽

Yoshiaki Tabuchi ◽

Ryohei Ogawa ◽

...

Keyword(s):

Tumor Necrosis Factor ◽

Tumor Necrosis ◽

Death Receptor ◽

Death Receptor 5 ◽

Osteosarcoma Cells ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Induced Apoptosis ◽

Death Receptor 4 ◽

Necrosis Factor

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Rhotekin 2 silencing inhibits proliferation and induces apoptosis in human osteosarcoma cells

Bioscience Reports ◽

10.1042/bsr20181384 ◽

2018 ◽

Vol 38 (6) ◽

Author(s):

Xiong Wang ◽

Lei Zhang ◽

Wenji Wang ◽

Yuchen Wang ◽

Ye Chen ◽

...

Keyword(s):

Cell Lines ◽

Phase Cell ◽

Osteosarcoma Cell ◽

Cyclin Dependent Kinase ◽

Potential Therapeutic Target ◽

Human Osteosarcoma ◽

Cycle Arrest ◽

Human Osteosarcoma Cells ◽

Human Osteosarcoma Cell ◽

Induced Apoptosis

Human osteosarcoma is the most frequent primary malignant of bone, and often occurs in adolescents. However, molecular mechanism of this disease remains unclear. In the present study, we found that the level of Rhotekin 2 (RTKN2) was up-regulated in osteosarcoma tissues and cell lines. In addition, silencing of RTKN2 of human osteosarcoma cell lines U2OS, inhibited proliferation, and induced G1 phase cell cycle arrest via reducing the level of the cyclin-dependent kinase 2 (CDK2). Furthermore, RTKN2 knockdown in the U2OS cells induced apoptosis by increasing the level of Bax and decreasing the level of Bcl2. These results suggested that RTKN2 is involved in the progression of human osteosarcoma, and may be a potential therapeutic target.

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HDAC6 Inhibitor WT161 Performs Antitumor Effect on Ostersarcoma and Synergistically Interacts with 5-FU

10.21203/rs.3.rs-49194/v1 ◽

2020 ◽

Author(s):

Jun Sun ◽

Xiaofeng Tang ◽

Feifei Zhang ◽

Cheng Ju ◽

Renfeng Liu ◽

...

Keyword(s):

Osteosarcoma Cell ◽

Dependent Manner ◽

Osteosarcoma Cells ◽

Proliferative Effect ◽

Xenograft Models ◽

Hdac6 Inhibitor ◽

Underlying Mechanisms ◽

Induced Apoptosis

Abstract Background: WT161 as a new selective HDAC6 inhibitor has been shown to play anti-tumor effects on multiple myeloma and breast cancer. However, the role of WT161 in osteosarcoma remains unclear. The aim of this study is to explore the role of WT161 in osteosarcoma and its underlying mechanisms.Methods: The anti-proliferative effect of WT161 on osteosarcoma cells was examined using MTT assay and colony formation assay. Cell apoptosis was analyzed using flow cytometer. The synergistic effect was evaluated by isobologram analysis using CompuSyn software. The osteosarcoma xenograft models were esatablished to evaluate the anti-proliferative effect of WT161 in vivo.Results: WT161 suppressed the cell growth and induced apoptosis of osteosarcoma cells in a dose- and time-dependent manner. Mechanistically, we found that WT161 treatment obviously increased the protein expression level of PTEN and decreased the phosphorylation level of AKT. Notably, WT161 shows synergistically inhibitory effects on osteosarcoma cell combined with 5-FU. Animal experiment results show WT161 inhibits the growth of osteosarcoma tumor and further illustrates that WT161 and 5-FU have a synergistic efficiency in osteosarcoma.Conclusions: These results indicate that WT161 inhibiting the growth of osteosarcoma through PTEN and has a synergistic efficiency with 5-FU.

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Mechanism of histone deacetylase inhibitor Trichostatin A induced apoptosis in human osteosarcoma cells

APOPTOSIS ◽

10.1023/b:appt.0000038037.68908.6e ◽

2004 ◽

Vol 9 (5) ◽

pp. 583-589 ◽

Cited By ~ 40

Author(s):

M. S. Roh ◽

C. W. Kim ◽

B. S. Park ◽

G. C. Kim ◽

J. H. Jeong ◽

...

Keyword(s):

Histone Deacetylase ◽

Histone Deacetylase Inhibitor ◽

Trichostatin A ◽

Osteosarcoma Cells ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Deacetylase Inhibitor ◽

Histone Deacetylase Inhibitor Trichostatin ◽

Induced Apoptosis

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11-O-Galloyl Bergenin from Corylopsis coreanas Leaves Induces Autophagy and Apoptosis in Human Osteosarcoma

The American Journal of Chinese Medicine ◽

10.1142/s0192415x21500956 ◽

2021 ◽

Vol 49 (08) ◽

pp. 2017-2031

Author(s):

Kyung-Ran Park ◽

Yoon-Ju Kwon ◽

Myounglae Cho ◽

Il Keun Kwon ◽

Jin Tae Hong ◽

...

Keyword(s):

Annexin V ◽

Mg63 Cell ◽

Underlying Mechanism ◽

Mapk Signaling ◽

Migration And Invasion ◽

Boyden Chamber ◽

Antitumor Effects ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Induced Apoptosis

Osteosarcoma is the most common malignant bone-forming tumor, wherein most patients with high grade osteosarcomas are treated with chemotherapy. Despite this, survival for metastatic or relapsed osteosarcoma patients has remained at an overall 5-year survival rate of 20%. In particular, the extracts of Corylopsis coreana (Korean winter hazel), a cultivated woody plant in South Korea, have shown beneficial anti-inflammatory, anti-oxidative, anti-osteoclastic, and antihyperuricemic properties. Therefore, this study aimed to demonstrate the antitumor activities and underlying mechanism of 11-O-Galloyl bergenin (OGAL) isolated from Corylopsis coreanas leaves in human osteosarcoma cells. Herein, we found that OGAL inhibited MG63 cell proliferation and induced cellular apoptosis as evidenced by cleaved-PARP, cleaved-caspase 3, TUNEL-positive cells, and Annexin V-positive cells. Specifically, OGAL-induced apoptosis was accompanied by p53 and p21 upregulation, BAX expression, and decreased Bcl-2 and cdk2. Moreover, OGAL induced autophagy via AKT inactivation, LC3II upregulation, and MG63 cell autophagosome formation. OGAL-induced autophagy was also accompanied by increased p38 phosphorylation, whereas JNK and ERK1/2 activities were found to be unaffected upon examining the MAPK signaling pathway. Furthermore, wound healing and Boyden chamber assays showed that OGAL suppressed MG63 cell migration and invasion. Given these findings, this study provided evidence that OGAL has antitumor effects by apoptosis and autophagy enhancement through increased p53, AKT, and p38 signaling, suggesting that OGAL may be a potential therapeutic strategy for osteosarcoma treatment.

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