Shikonin derivatives cause apoptosis and cell cycle arrest in human chondrosarcoma cells via death receptors and MAPK regulation

Abstract

Background Although chondrosarcoma is the second most common primary malignant bone tumor, treatment options are limited due to its extensive resistance to a chemo- and radiation therapy. Since shikonin has shown potent anticancer activity in various types of cancer cells, it represents a promising compound for the development of a new therapeutic approach. Methods The dose-relationships of shikonin and its derivatives acetylshikonin and cyclopropylshikonin on two human chondrosarcoma cell lines were measured using the CellTiter-Glo®. The changes in the cell cycle were presented by flow cytometry. Protein phosphorylation and expression apoptotic markers, MAPKs and their downstream targets were analyzed using western blotting and gene expression were evaluated using RT-qPCR. Results Chondrosarcoma cells showed a dose-dependent inhibition of cell viability after treatment with shikonin and its derivatives, with the strongest effect for shikonin and IC50 values of 1.3 ± 0.2 µM. Flow cytometric measurements revealed a G2/M arrest of the cells after treatment. Protein and gene expression analysis demonstrated a dose-dependent downregulation of survivin and XIAP, and an upregulation of Noxa, γH2AX, cleaved caspase-8, -9, -3, and -PARP. Furthermore, the expression of various death receptors was modulated. As MAPK signaling pathways play a key role in tumor biology, their phosphorylation pattern and their corresponding downstream gene regulation were analyzed. Treatment with shikonin derivatives caused an inhibition of pSTAT3 and an increase of pAKT and the MAPKs pERK, pJNK, and pp38 in a dose-dependent manner. Conclusions These data demonstrated the significant anti-tumorigenic effect of shikonin derivatives in chondrosarcoma and encourage further research.

Acetylshikonin induces necroptosis via the RIPK1/RIPK3-dependent pathway in lung cancer

“…Note that this was confirmed by assessing the expression of cell cycle regulatory proteins via western blot analysis. Interestingly, these findings are consistent with those in previous studies on chondrosarcoma cells [ 34 ], but not in studies on colorectal cancer, in which cell cycle arrest occurred in the G1 phase [ 52 ]. These inconsistencies pertaining to the anticancer effects of acetylshikonin deserve further consideration in the context of lung cancer.…”

“…Research has shown that acetylshikonin induces apoptosis in hepatocellular carcinoma and oral squamous cell carcinoma by triggering the production of intracellular reactive oxygen species [ 32 , 33 ]. Acetylshikonin treatment has also been shown to promote cell cycle arrest in the G2/M and S phases in chondrosarcoma and leukemia cells [ 34 , 35 ]. Functional kinetics analysis suggests that the blockading of cellular drug transporters can be attributed to acetylshikonin increasing the sensitivity of multidrug-resistant human gastric and breast cancer cells to chemotherapy drugs [ 36 ].…”

Shikonin reactivates TSGs GADD45B and PPP3CC to block NSCLC cell proliferation and migration through JNK/P38/MAPK signaling pathways

“…The main molecular function of the top upregulated genes in KEGG pro ling was the MAPK signaling pathway shown in Fig. 3C, which was consistent with the ndings of previous studies on the shikonin-induced activation of the MAPK pathway (21,22).…”

Temozolomide, Simvastatin and Acetylshikonin Combination Induces Mitochondrial Dependent Apoptosis in GBM Cells which is Regulated by Autophagy

“…It has also been reported that ASH induces dose-dependent apoptosis via activation of caspase-3/-7 and -9 in chondrosarcoma cell lines [88]. They also showed that MAPK activation is involved in ASHinduced apoptosis in colorectal cancer cells [88]. On the other hand, ASH induces caspasedependent apoptosis via ROS and inhibition of NF-κB in K562 leukemia cells [89].…”

“…Recent investigations have shown that ASH increases ROS and nuclear damage with subsequent nuclear translocation of FOXO3 and induced caspase-dependent apoptosis in osteosarcoma U2OS, renal cell carcinoma and colorectal cancer HCT-15, and LoVo cells [85][86][87]. It has also been reported that ASH induces dose-dependent apoptosis via activation of caspase-3/-7 and -9 in chondrosarcoma cell lines [88]. They also showed that MAPK activation is involved in ASHinduced apoptosis in colorectal cancer cells [88].…”