Effect of dihydroartemisinin on invasion , migration , and apoptosis of neuroma cells by regulating the CX 3 CR 1 signaling pathway

Objective: To elucidate mechanisms by which dihydroartemisinin (DHA) induces the apoptosis of neuroma cells by regulating the CX3CR1 signaling pathway. Methods: The U251 cell line was cultured in vitro and randomly assigned to the following 6 groups after transfection and treatment: the Blank group (controls), the DHA (50 μmol/L) group (with low-concentration DHA), the DHA (75 μmol/L) group (with moderate-concentration DHA), the DHA (100 μmol/L) group (with high-concentration DHA), the si-CX3CR1 group, and the DHA (100 μmol/L) + si-CX3CR1 group. qRT-PCR and Western blot analysis were conducted to detect the mRNA and protein levels of CX3CR1, the upstream ligand CX3CL1, and the downstream AKT. The apoptotic signals Caspase-3, Bcl-2, and FADD, were also detected along with activated p-AKT. The CCK-8 assay was conducted to determine changes in cell proliferation. Wound healing and transwell assays were performed to detect migration and invasion profiles of transfected cells. Flow cytometry analysis was done to examine the apoptosis profile. Results: As compared to the Blank group, the mRNA and protein expression of CX3CL1, CX3CR1, Bcl-2, and AKT declined. Whereas p-AKT expression declined, mRNA and protein expression of Caspase-3 and FADD rose substantially in the DHA (50 μmol/L) group, the DHA (75 μmol/L) group, the DHA (100 μmol/L) group, and the si-CX3CR1 group (all P<0.05). With the rise of DHA concentrations, the mRNA and protein expression of CX3CL1, CX3CR1, Bcl-2 and AKT were decreased more significantly, while Caspase-3 mRNA and protein expression were increased more remarkably. When compared to the Blank group, proliferation, invasion and migration of cells at different time points were attenuated in the DHA (50 μmol/L) group, the DHA (75 μmol/L) group, the DHA (100 μmol/L) group, and the si-CX3CR1 group, but the apoptosis rates became higher (all P<0.05). With the increase in DHA concentration, proliferation, invasion, and migration of cells at distinct time points were inhibited, and apoptosis rates were increased sequentially (all P<0.05). The apoptotic rates of cells in the Blank group, the DHA (50 μmol/L) group, the DHA (75 μmol/L) group, the DHA (100 μmol/L) group, the si-CX3CR1 group, and the DHA (100 μmol/L) + si-CX3CR1 group were (13.14±1.23)%, (32.95±3.26)%, (48.52±4.21)%, (62.35±6.12)%, (65.57±7.52)% and (64.52±6.85)%, respectively. Conclusion: DHA can inhibit invasion and migration of neuroma cells by inhibiting the CX3CR1 signaling pathway, thereby promoting apoptosis of neuroma cells.

[1]  Y. Li,et al.  Dihydroartemisinin increases apoptosis of colon cancer cells through targeting Janus kinase 2/signal transducer and activator of transcription 3 signaling. , 2017, Oncology letters.

[2]  Yang Li,et al.  CX3CR1 regulates osteoarthrosis chondrocyte proliferation and apoptosis via Wnt/β-catenin signaling. , 2017, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[3]  Jian Zheng,et al.  Dihydroartemisinin Exerts Anti-Tumor Activity by Inducing Mitochondrion and Endoplasmic Reticulum Apoptosis and Autophagic Cell Death in Human Glioblastoma Cells , 2017, Front. Cell. Neurosci..

[4]  Hui Wang,et al.  Fractalkine/CX3CR1 induces apoptosis resistance and proliferation through the activation of the AKT/NF‐κB cascade in pancreatic cancer cells , 2017, Cell biochemistry and function.

[5]  P. Cai,et al.  Dihydroartemisinin inhibits the viability of cervical cancer cells by upregulating caveolin 1 and mitochondrial carrier homolog 2: Involvement of p53 activation and NAD(P)H:quinone oxidoreductase 1 downregulation , 2017, International journal of molecular medicine.

[6]  D. McTigue,et al.  Deletion of the Fractalkine Receptor, CX3CR1, Improves Endogenous Repair, Axon Sprouting, and Synaptogenesis after Spinal Cord Injury in Mice , 2017, The Journal of Neuroscience.

[7]  Mingjun Wu,et al.  Mechanism of dihydroartemisinin-induced apoptosis in prostate cancer PC3 cells: An iTRAQ-based proteomic analysis. , 2016, Life sciences.

[8]  M. Amit,et al.  Mechanisms of cancer dissemination along nerves , 2016, Nature Reviews Cancer.

[9]  Yu-qi Fan,et al.  Artemisinin inhibits monocyte adhesion to HUVECs through the NF-κB and MAPK pathways in vitro , 2016, International journal of molecular medicine.

[10]  Yun Zhang,et al.  Novel Small-Molecule CX3CR1 Antagonist Impairs Metastatic Seeding and Colonization of Breast Cancer Cells , 2016, Molecular Cancer Research.

[11]  I. Kurkowska-Jastrzębska,et al.  Analysis of the Role of CX3CL1 (Fractalkine) and Its Receptor CX3CR1 in Traumatic Brain and Spinal Cord Injury: Insight into Recent Advances in Actions of Neurochemokine Agents , 2016, Molecular Neurobiology.

[12]  Carla M. T. Bauer,et al.  Self-renewing resident arterial macrophages arise from embryonic CX3CR1+ precursors and circulating monocytes immediately after birth , 2015, Nature Immunology.

[13]  Chengcheng Guo,et al.  Dihydroartemisinin increases temozolomide efficacy in glioma cells by inducing autophagy. , 2015, Oncology letters.

[14]  Jie Li,et al.  Dihydroartemisinin inhibits endothelial cell proliferation through the suppression of the ERK signaling pathway. , 2015, International journal of molecular medicine.

[15]  Q. Shi,et al.  Effects of expression silencing of chemokine receptor CX3CR1 on human hepatocellular carcinoma Huh7 cells and its machanism , 2015 .

[16]  Fan Wang,et al.  Dihydroartemisinin suppresses glioma proliferation and invasion via inhibition of the ADAM17 pathway , 2015, Neurological Sciences.

[17]  H. Feng,et al.  Dihydroartemisinin exhibits anti-glioma stem cell activity through inhibiting p-AKT and activating caspase-3. , 2014, Die Pharmazie.

[18]  X. Qu,et al.  Involvement of the mitochondrial pathway and Bim/Bcl-2 balance in dihydroartemisinin-induced apoptosis in human breast cancer in vitro. , 2013, International journal of molecular medicine.

[19]  J. Ning.  Study On the Relationship Among MMP-2,Atherosclerosis And Cerebral Infarction , 2012 .

[20]  M. Giustetto,et al.  Synaptic Pruning by Microglia Is Necessary for Normal Brain Development , 2011, Science.

[21]  Hyung Gyun Kim,et al.  Suppression of PMA-induced tumor cell invasion by dihydroartemisinin via inhibition of PKCalpha/Raf/MAPKs and NF-kappaB/AP-1-dependent mechanisms. , 2010, Biochemical pharmacology.

[22]  J. An,et al.  Dihydroartemisinin upregulates death receptor 5 expression and cooperates with TRAIL to induce apoptosis in human prostate cancer cells , 2010, Cancer biology & therapy.

[23]  P. Allavena,et al.  Molecular mechanisms of perineural invasion, a forgotten pathway of dissemination and metastasis. , 2010, Cytokine & growth factor reviews.

[24]  Ravi Salgia,et al.  Regulation of cellular proliferation, cytoskeletal function, and signal transduction through CXCR4 and c-Kit in small cell lung cancer cells. , 2002, Cancer research.

[25]  C. Combadière,et al.  Identification of CX3CR1. A chemotactic receptor for the human CX3C chemokine fractalkine and a fusion coreceptor for HIV-1. , 1998, The Journal of biological chemistry.

[26]  C. Combadière,et al.  Cloning, chromosomal localization, and RNA expression of a human beta chemokine receptor-like gene. , 1995, DNA and cell biology.