Microglia contribute to nociception via CSF-1R signaling pathway in rat orofacial carcinoma.

OBJECTIVE: Cancer-induced pain is the most common complication of the head and neck cancer. The microglia colony-stimulating factor receptor 1 (CSF1R) plays a crucial role in the inflammation and neuropathic pain. However, the effect of CSF1R on orofacial cancer-induced pain is unclear. Here, we aimed to determine the role of CSF1R in orofacial pain caused by cancer. METHODS: We established an animal model of cancer-induced orofacial pain with Walker 256B cells. Von Frey filament test and laser-intensity pain tester were used to evaluate tumor-induced mechanical and thermal hypersensitivity. Minocycline and PLX3397 were used to alter tumor-induced mechanical and thermal hyperalgesia. Additionally, we evaluated the effect of PLX3397 on immunoinflammatory mediators and neuronal activation within the trigeminal spinal subnucleus caudalis (Vc). RESULTS: Walker 256B cell-induced tumor growth resulted in mechanical and thermal hyperalgesia, accompanying by microglia activation and CSF1R upregulation. Treatment with minocycline or PLX3397 reversed the associated nocifensive behaviors and microglia activation triggered by tumor. As a result of PLX3397 treatment, tumor-induced increases in pro-inflammatory cytokine expression and neuronal activation of the Vc were significantly inhibited. CONCLUSIONS: The results of our study showed that blocking microglial activation via CSF1R may help prevent cancer-induced orofacial pain.

[Changes and significance of serum prealbumin expression in patients with oral and maxillofacial space infections].

PURPOSE: To investigate the changes of serum prealbumin (PA) expression level in patients with oral and maxillofacial space infections and its significance. METHODS: Patients hospitalized at the Affiliated Hospital of Xuzhou Medical University from January 2020 to September 2021 were selected and divided into infected and non-infected group. One hundred and twenty-one patients with moderate to severe oral and maxillofacial gap infections were in the infected group, and 128 patients without infection were in the non-infected group. In the infected group, PA, high-sensitivity C-reactive protein (hs-CRP) and white blood cell count (WBC) levels and related clinical parameters were measured at 1, 3 and 7 d of admission. In the non-infected group, PA, hs-CRP and WBC levels were measured at 1 d of admission. SPSS 23.0 software package was used to statistically analyze the relationship between PA levels and various laboratory and clinical parameters. RESULTS: PA levels in the infected group were significantly lower than those in the non-infected group at 1 d of admission. PA levels in the infected group showed an overall increasing trend at different time points, and PA was negatively correlated with pain intensity and positively correlated with mouth opening(P＜0.05). The diagnostic sensitivity was 90.91% and the specificity was 92.97% for PA≤19.85 mg/dL, which can be used as the best diagnostic threshold. The diagnostic efficacy can be improved when combined with hs-CRP and WBC. Logistic regression analysis showed that low PA level was an independent risk factor for patients requiring intensive care after surgery (P＜0.05). CONCLUSIONS: PA is an effective tool for the early diagnosis and evaluation of the efficacy of oral and maxillofacial interstitial infections, and can be used as a reference indicator to assess prognosis.

[Artesunate inhibited human tongue squamous cell carcinoma CAL27 cells in vitro via STAT3 signaling pathway].

PURPOSE: To investigate the anticancer effect of artesunate(ART) on human tongue squamous cell carcinoma (CAL27) cells and its possible mechanism. METHODS: CAL27 cells was pretreated with different doses of ART. Then, CCK-8 and colony forming methods were used for cell viability analysis, cell apoptosis was detected by flow cytometry, and cell migration and invasion capacity were determined by scratch test and Transwell chamber method. In addition, the expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) mRNA was measured by qPCR, and Western blot was used to detect the expression of MMP-9 and VEGF and activation of STAT3 signal in CAL27 cells treated with ART at various concentrations. SPSS 19.0 software package was used for statistical analysis of the data. RESULTS: Artesunate significantly inhibited proliferation(P＜0.01), invasion(P＜0.01)and migration(P＜0.01) of CAL27 cells，and induced apoptosis of CAL27 cells (P＜0.01) in a dose-dependent manner. ART not only significantly reduced the expression of MMP-9 and VEGF mRNA in CAL27 cells in a dose-dependent manner(P＜0.05), but also inhibited the protein expression of p-STAT3, MMP-9 and VEGF. CONCLUSIONS: Artesunate can inhibit the proliferation, migration and invasion of CAL27 cells, which may exert antitumor effects by inhibiting the STAT3 signaling pathway.

Role of miR-21 in the growth and metastasis of human salivary adenoid cystic carcinoma.

Aberrant microRNA (miRNA/miR) expression has been reported in various cancer types. miR­21, which is considered to be a proto-oncogene and is frequently overexpressed in certain cancer types, has been implicated in tumorigenesis. The aim of the present study was to investigate the effect of miR­21 degradation on tumor progression and its potential mechanisms in human salivary adenoid cystic carcinoma (SACC) development. Results of reverse transcription­quantitative polymerase chain reaction analysis indicated that SACC cells with high metastatic potential (SACC­LM cells) exhibited a significantly higher expression of miR­21 compared with SACC cells with a lower metastatic potential (SACC­83 cells). In addition, following transfection of SACC­LM cells with miR­21 inhibitor, cell viability was reduced, which may be a result of reduced cell proliferation and metastasis, and the induction of apoptosis, as determined by Cell Counting Kit­8, wound healing, Matrigel invasion and flow cytometry assays. Furthermore, bioinformatics analysis indicated that programmed cell death 4 (PDCD4), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and B­cell lymphoma (Bcl)­2 are potential target genes of miR­21. Therefore, western blotting was performed to investigate the expression of these proteins, and the results demonstrated that miR­21 expression level was negatively associated with PDCD4 and PTEN protein expression, and positively associated with Bcl­2 protein expression, in SACC­LM cells, indicating that miR­21 may promote SACC progression via PDCD4, PTEN and Bcl­2. In conclusion, the present study indicates that miR­21 may be a novel target for SACC therapy and provide a novel basis for the clinical treatment of SACC.

Effect of survivin downregulation by simvastatin on the growth and invasion of salivary adenoid cystic carcinoma.

Simvastatin, an inhibitor of 3­hydroxy­3-methylglutaryl­coenzyme A reductase, is been used in the clinic due to its pleiotropic effects, such as breast cancer, prostate cancer, pancreatic cancer. Simvastatin has recently been demonstrated to serve a potential role in the prophylaxis and therapeutics of a number of human cancers. The majority of reports concerning simvastatin treatment in the majority of human cancers have demonstrated that survivin is significantly decreased as a result and has been implicated in tumorigenesis. However, only a limited number of studies have investigated the use of simvastatin for the treatment of salivary gland adenoid cystic carcinoma (SACC). Therefore, this agent is a candidate for further investigation. The aim of the present study was to investigate the effects of simvastatin on the proliferation, invasion and apoptosis of the human salivary adenoid cystic carcinoma cell line, SACC­83, as well as survivin expression in the cells. The Cell Counting kit­8 assay results revealed that simvastatin inhibited the proliferation of SACC­83 cells in a dose­dependent (10 to 50 µM) and time­dependent (24 to 48 h) manner when compared with the untreated cells. Flow cytometry analysis indicated that simvastatin increased the percentage of cells in early and late apoptosis. Invasion assays revealed that simvastatin treatment inhibited the invasiveness of SACC­83 cells in a dose­dependent manner. In addition, simvastatin downregulated survivin expression in SACC­83 cells. In conclusion, simvastatin significantly inhibited the proliferation and invasion of SACC­83 cells, induced apoptosis, and reduced the expression of survivin, which suggests that simvastatin may be a novel target for SACC therapy.

Effect of simvastatin and microRNA-21 inhibitor on metastasis and progression of human salivary adenoid cystic carcinoma.

Salivary adenoid cystic carcinoma (SACC) is a common malignancy of the salivary glands. Epithelial-mesenchymal transition (EMT) and P53 signaling pathway are associated with SACC metastasis and progression. Although simvastatin (SIM) is effective against the growth of many cancer types, its side effects limit its use. microRNA-21 (miR-21) is highly expressed in a variety of tumors and has a role in promoting tumor development. Therefore, the aim of the present study was to evaluate the effect of SIM in combination with miR-21 inhibitor (miR-21i) against lung metastatic SACC cells (SACC-LM). Our results showed that miR-21i was effective in reducing the resistance of SACC-LM to SIM, resulting in SACC-LM acquisition of epithelial traits, cell migration and invasion reduction, growth inhibition and induction of apoptosis. The expression of proteins associated to metastasis and tumor progression were regulated by the combined use of SIM and miR-21i. Thus, our findings demonstrated that such combination was effective in inhibiting SACC-LM progression, suggesting that multi-target therapy against SACC might represent a potentially successful approach in clinical treatment.

MiR-34a inhibits oral cancer progression partially by repression of interleukin-6-receptor.

Previous reports revealed that a significant decrease of miR-34a in oral cancer. But the role of miR-34a in oral cancer needs further research. In the present study, we will investigate the effect of miR-34a on oral cancer cell phenotypes. First, it was verified that miR-34a expression was lower in oral cancer tissues compared with their normal controls, so did the oral cancer cells. Next, it was showed that miR-34a overexpression in oral cancer cells could inhibit cell proliferation, G1 phase arrest, metastasis and epithelial mesenchymal transition. It was predicted that interleukin-6-receptor (IL6R) was a potential target gene of miR-34a by bioinformatics analysis and identified by luciferase assay. It was further showed that miR-34a inhibited oral cancer progression via IL6R. Collectively, our findings suggested that miR-34a may function as a tumor suppressor in oral cancer by targeting IL6R.

[Effects of dexamethasone and vitamin B12 on EGF, TGFalpha, and TGFbeta1 gene expressions of A/J mouse palatal cells in vitro].

OBJECTIVE: To investigate the molecular mechanism of congenital cleft palate induced by Dexamethasone (DEX) and the counteractive effect of Vitamin B12. METHODS: The RT-PCR was used to detect the independent and joint effects of DEX and Vitamin B12 on the gene expressions of A/J embryonic mouse palatal cells. RESULTS: DEX stimulated the gene expressions of EGF and TGFbeta1, while the expression of TGFalpha left unchanged in the palatal cells. Vitamin B12 had no direct effects on the gene expressions of EGF, TGFalpha, and TGFbeta1; however, it counteracted the effects of DEX on the palate cells. CONCLUSION: DEX induces cleft palate through increasing some gene expressions of growth factors, which can be inhibited by Vitamin B12.

[Influence of dexamethasone on fusion of embryonic palatal medial edge epithelium in mouse palatal shelves in vitro].

OBJECTIVE: To study the effect of dexamethasone on the differentiation and proliferation of type A mouse palatal medial edge epithelial cells when there is type A mouse embryonic palatal mesenchymal cells. METHODS: The mouse palatal shelves were harvested from a female mouse of gestation day 14 by microsurgical dissection and cultured in vitro. The differentiation was investigated through microscope and transmission electron microscope under condition of the palatal shelves fusion. RESULTS: Dexamethasone promoted the palatal medial edge epithelium differentiated into squamause epithelium and affected normal development and obstructed the fusion of mouse palatal shelves. CONCLUSION: The results of histological observation indicate that dexamethasone promotes the proliferation of palatal meseuchymal cells and inhibits the normal differentiation of palatal medial edge epithelial cells, which results in cleft palate.

[A cephalometric study of the craniofacial morphology in adult patients with unoperated cleft palate.].

PURPOSE: To evaluate the effects of cleft palate on the craniofacial morphology in adult patients with unoperated cleft palate by cephalometric analysis. METHODS: X-ray cephalometry was used for the study of craniofacial morphology carried out in 7 male and 8 female patients with unoperated cleft palate. Pointing measurements were done with the methods of Downs and Steiner. The data of the cephalometric measurements were compared with that of Guangxi native normal adults by t test. RESULTS: The unoperated cleft palate patient,s SNA,ANS-Ptm(FH) and L1-MP were smaller or shorter than those of normal group while Po-NB was longer. There was significant difference between the two groups (P<0.05 or 0.01). CONCLUSIONS: The deformities of adult patients with unoperated cleft palate were mainly considered to be that the anterior and posterior length of maxilla show hypoplastic while the vertical height of the maxilla the length of anterior cranial fossa and mandible was normal. But the retrusion of alveolar mandible led to chin protrusion.