Dysregulated miR-645 affects the proliferation and invasion of head and neck cancer cell.

BACKGROUND AND PURPOSE: Dysregulated miRNAs play an important role in many malignant tumors. However, elucidating the roles of miRNAs in cancer biology, especially in epithelial cancers, remains an ongoing process. In this study, we identified the differentially expressed miR-645 in the progressing of head and neck squamous cell carcinoma (HNSCC) and investigated its biological function. METHODS: The association between clinicopathological parameters and the expression levels of the candidated miRNAs were analyzed by using the Kaplan-Meier survival analysis. The cell growth, invasion and migration potential, and clone formation were observed to detect the functions of the miRNAs in HNSCC cells. RESULTS: In the 34 HNSCC tissues with lymph node metastasis, the expression level of miR-645 was 0.54 ± 0.12, and the expression level was 0.22 ± 0.05 in the 28 tissues with non lymph node metastasis (p = 0.017). In patients with HNSCC, higher level of miR-645 expression significantly correlates with worse overall survival (p = 0.04). Ectopic expression of miR-645 promoted cell invasion and migration. CONCLUSIONS: miR-645 play a key role in cell invasion and metastasis and their expression correlates with overall survival in the patients with HNSCC.

Extracellular vesicles secreted by human periodontal ligament induced osteoclast differentiation by transporting miR-28 to osteoclast precursor cells and further promoted orthodontic tooth movement.

BACKGROUND: Osteoclast differentiation plays a key role in orthodontic tooth movement (OTM). We aimed to explore the role of human periodontal ligament (hPDL) extracellular vesicles (EVs) in osteoclast differentiation and OTM. METHODS: The hPDL cells were exposed to 4.0 g/cm2 compression force (CF) and the hPDL-EVs were collected. The peripheral blood mononuclear cells were isolated, purified, and induced osteoclast differentiation. The OTM rat model was established through excess orthodontic force. Dual-luciferase reporter gene assay verified the targeting effect of miR-28 on RUNX1. In addition, tartrate-resistant acid phosphase (TRAP) staining, immunofluorescence, western blot, and quantitative real-time PCR were also carried out. RESULTS: CF pretreated hPDL-EVs promoted osteoclast differentiation and down-regulated RUNX1 levels in in vitro and in vivo experiments. The addition of CF-hPDL-EVs also elevated tooth movement in OTM rats. Besides, miR-28 was significantly up-regulated in CF-pretreated hPDL-EVs. In addition, RUNX1 was negatively regulated by miR-28. Moreover, the addition of CF-lenti-miR-28 inhibitor-Evs down-regulated the expression of osteoclast marker genes and the number of TRAP positive (+) multinucleated cells (MNCs) in vitro. Furthermore, in vivo experiments confirmed that CF-lenti-miR-28 inhibitor-Evs injection down-regulated the number of TRAP (+) MNCs and inhibited tooth movement of OTM rats. CONCLUSION: CF-treated hPDL-EVs promoted osteoclast differentiation by transporting miR-28 and inhibiting the expression of RUNX1, which provides new insight into the specific mechanism of hPDL-Evs affecting osteoclast differentiation.

Erratum: HIF-1α or HOTTIP/CTCF Promotes Head and Neck Squamous Cell Carcinoma Progression and Drug Resistance by Targeting HOXA9.

[This corrects the article DOI: 10.1016/j.omtn.2019.12.045.].

HIF-1α or HOTTIP/CTCF Promotes Head and Neck Squamous Cell Carcinoma Progression and Drug Resistance by Targeting HOXA9.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most frequently diagnosed cancer worldwide. However, the clinical outcomes remain unsatisfactory. The aim of this study is to unravel the functional role and regulatory mechanism of HOXA9 in HNSCC. A cohort of 25 HNSCC tumor tissues and normal tissue counterparts was collected. qRT-PCR and western blotting were performed to determine the levels of HOXA9 and epithelial-mesenchymal transition (EMT)-related markers. Cell Counting Kit-8 (CCK-8) and colony formation assays were conducted to monitor cell viability and cytotoxicity. Transwell and wound healing assays were used to determine cell migration and invasion. Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) staining was performed to detect cell apoptosis. Bioinformatic analysis, electrophoretic mobility shift assay and chromatin immunoprecipitation (ChIP) assays were performed to investigate the direct binding between HIF-1α or CCCTC binding factor (CTCF) and HOXA9. Glutathione S-transferase (GST) pull-down and RNA pull-down assays were used to validate the interaction between CTCF and HOTTIP. HOXA9 was upregulated in HNSCC tissues and cells. Knockdown of HOXA9 inhibited cell proliferation, migration, invasion, and chemoresistance but promoted apoptosis in CAL-27 and KB cells. Knockdown of HOXA9 also regulated EMT-related marker via targeting YAP1/ß-catenin. Silencing of HOTTIP or CTCF exerted similar tumor-suppressive effects in HNSCC. Mechanistically, HIF-1α or CTCF transcriptionally regulated HOXA9, and HOTTIP/CTCF cooperatively regulated HOXA9 in KB cells. HIF-1α or HOTTIP/CTCF transcriptionally modulates HOXA9 expression to regulate HNSCC progression and drug resistance.

Knockdown of DANCR reduces osteoclastogenesis and root resorption induced by compression force via Jagged1.

LncRNA DANCR has been proven to be involved in osteoblast differentiation. This study aims to investigate the role of DANCR in osteoclast formation and root resorption in periodontal ligament (PDL) cells induced by compression force (CF). Rat orthodontic tooth movement (OTM) model was established. The molecules expressions in the areas of root resorption form OTM model were measured. The number of osteoclasts was measured using Tartrate-resistant acid phosphatase (TRAP) staining. The bone resorption was detected using pit formation assay. We showed that the expression of DANCR and Jagged1 protein was increased in rat OTM model and human periodontal ligament (hPDL) cells treated with CF, and CF increased the production of Jagged1, RANKL, and IL-6 from the hPDL cells. Moreover, DANCR could positively regulate Jagged1 protein expression. Knockdown of DANCR could change the promotion effect of CF on osteoclastogenesis and bone resorption in vitro and in vivo experiments, while overexpression of Jagged1 reversed si-DANCR effect. Taken together, knockdown of DANCR reduced osteoclast formation and root resorption induced by CF via Jagged1.

Lactoferricin B reverses cisplatin resistance in head and neck squamous cell carcinoma cells through targeting PD-L1.

Head and neck squamous cell carcinoma (HNSCC) ranks among the top most common cancers with a poor prognosis. The mechanism of chemoresistance is still not well known. This study is to investigate the programmed death-ligand 1 (PD-L1) expression in HNSCC, and test the effect of lactoferricin B (LfcinB) on chemoresistance and its mechanism. We analyzed 510 HNSCC patients in TCGA database and investigated how CD274 expression was related to patient prognosis. PD-L1 was verified from HNSCC samples at local hospital with immunohistochemistry. PD-L1 expression in the acquired cisplatin-resistant HNSCC cells was examined by PCR and WB in order to test PD-L1-induced chemoresistance. LfcinB inoculation in cisplatin-resistant HNSCC cells and in the nude mice was introduced to test the effect of LfcinB on targeting cisplatin resistance and its mechanism. High CD274 mRNA (>125 FPKM) from TCGA database had a significantly reduced 5-year survival rate, and a lower 5-year survival rate in the chemotherapy and radiotherapy-treated patients (P < .05). PD-L1 overexpression was further supported from analysis of 40 HNSCC specimens. PD-L1 and IL-6 in the established cisplatin-resistant HNSCC cells were shown significantly higher (P < .05). IL-6 and PD-L1 expression were partially inhibited by the anti-IL-6/STAT3 antibody. LfcinB displayed a direct cytotoxic effect on cisplatin-resistant HNSCC cells and HNSCC xenografts of cisplatin-resistant cells in the nude mice displayed significant reduction in tumor volume after LfcinB injection (P < .05). Besides, the increase of IL-6 and PD-L1 in cisplatin-resistant HNSCC cells was abolished in vitro by LfcinB (P < .05). PD-L1 expression in HNSCC cells correlates with poor prognosis and chemoresistance, and LfcinB might provide therapeutic potential in HNSCC patients through modulating IL-6 and PD-L1.

Hemangiopericytoma of the thoracic spine: a case report.

Hemangiopericytoma (HPC) has been described to be aggressive and potentially a malignant tumour. We report a rare case of a 63-year-old Chinese male who presented with primary intradural extramedullary HPC of the thoracic spine. The main presenting complaint was gradual progression of back pain, associated with paraparesis and sensory deficit of lower limbs. He had MRI thoracolumbar with contrast which showed T9 lesion compressing on spinal cord and oedema, he was then operated upon and histopathology report confirmed a thoracic spine HPC. A T8/9 laminectomy and excision of intradural extramedullary lesion was performed, tumour section was sent for frozen section study, and more tissue was sent for paraffin studies and additional immunohistochemical staining. Surgical resection is most commonly performed, radiotherapy remains debatable. In this report, we discussed another rare case of primary spinal HPC to be added into the literature.

Icaritin induces mitochondrial apoptosis by up-regulating miR-124 in human oral squamous cell carcinoma cells.

AIM OF THE STUDY: The present study is aimed to investigate the apoptosis-inducing effect of icaritin in human oral squamous cell carcinoma (OSCC) cells and the associated mechanisms. MATERIALS AND METHODS: KB and SCC9 cell lines were used as model cell lines. Effect of icaritin on apoptosis was analyzed by flow cytometry. The effect of icaritin on mitochondrial apoptotic pathway was demonstrated by loss of mitochondrial membrane potential and release of cytocrome C from mitochondria. MiR-124 mimic and miR-124 inhibitor were used to manipulate the expression of miR-124 in OSCC cells. SiRNA targeting Sp1 and DNMT1 as well as Sp1 and DNMT1 overexpressing vector were utilized to confirm their roles in the apoptosis-inducing effect of icaritin in OSCC cells. Activation of relevant signaling pathway by icaritin and effect of icaritin on expression of targeting molecules were determined by western blots or qRT-PCR. RESULTS: Our results showed that icaritin inhibited tumor cell viability in a dose- and time-dependent manner, and induced cell apoptosis via intrinsic mitochondrial pathway by upregulating miR-124. Moreover, our results showed that the icaritin exerted regulatory effect on miR-124 through suppressing Sp1/DNMT1 signaling. CONCLUSION: Our data provide the first experimental evidence that icaritin induces mitochondrial apoptosis in OSCC cells by upregulating miR-124 and suggest a new mechanism to explain its anti-tumor effects.

14-3-3ζ regulates immune response through Stat3 signaling in oral squamous cell carcinoma.

Ectopic expression of 14-3-3ζ has been found in various malignancies, including lung cancer, liver cancer, head and neck squamous cell carcinoma (HNSCC), and so on. However, the effect of 14-3-3ζ in the regulation of interactions between tumor cells and the immune system has not been previously reported. In this study, we aimed to investigate whether and how 14-3-3ζ is implicated in tumor inflammation modulation and immune recognition evasion. In oral squamous cell carcinoma (OSCC) cell lines and cancer tissues, we found that 14-3-3ζ is overexpressed. In OSCC cells, 14-3-3ζ knockdown resulted in the up-regulated expression of inflammatory cytokines. In contrast, 14-3-3ζ introduction attenuated cytokine expression in human normal keratinocytes and fibroblasts stimulated with interferon-γ (IFN-γ) and lipopolysaccharide (LPS). Furthermore, supernatants from 14-3-3ζ knockdown OSCC cells dramatically altered the response of peritoneal macrophages, dendritic cells and tumor-specific T cells. Interestingly, Stat3 was found to directly interact with 14-3-3ζ and its disruption relieved the inhibition induced by 14-3-3ζ in tumor inflammation. Taken together, our studies provide evidence that 14-3-3ζ may regulate tumor inflammation and immune response through Stat3 signaling in OSCC.

Reduced cytotoxicity of silver ions to mammalian cells at high concentration due to the formation of silver chloride.

Silver-containing antimicrobial agents are used in various medical products. However, their toxicity to mammalian cells has not been sufficiently evaluated. Numerous studies have unveiled evidence of significant antimicrobial properties associated with Ag ions. In cell culture media or human body fluids, the free Ag(+) has rich opportunities to complex with Cl(-). Surprisingly, studies on the toxicity of solid form AgCl(s) to mammalian cells are quite limited. In this study, we evaluated the cytotoxicity of Ag ions and silver chloride colloids on red blood cells and human mesenchymal stem cells (hMSCs). The adverse effects of silver chloride on red blood cells and hMSC were viewed by SEM and LIVE/DEAD viability staining, respectively. Among different tested chemical forms of silver, AgCl was identified to be the least cytotoxic. Moreover, a decline in the cytotoxicity of AgCl at significantly high concentrations was observed. We attributed the reduced cytotoxicity to aggregated AgCl which limited the bioavailability of free Ag(+) ions.

[Wrist joint reconstruction with vascularized fibular head graft after resection of distal radius giant cell tumor].

OBJECTIVE: To observe the effectiveness of wrist joint reconstruction with vascularized fibular head graft after resection of distal radius giant cell tumor. METHODS: Between March 2000 and March 2009, 31 cases of distal radius giant cell tumor were treated with extended resection and vascularized fibular head graft for repairing defects of the distal radius, and reconstructing wrist joint. There were 14 males and 17 females with an average age of 37.2 years (range, 15-42 years). The disease duration ranged from 1 month to 2 years and 3 months with an average of 8 months. The size of tumor was 6.5 cm x 3.5 cm-8.0 cm x 4.5 cm. The range of motion (ROM) of wrist joint was as follows: extension 5-15 degrees (mean, 10.7 degrees), flexion 9-21 degrees (mean, 14.2 degrees), radial inclination 0-10 degrees (mean, 8.6 degrees), and ulnar inclination 0-15 degrees (mean, 7.9 degrees). The ROM of forearm was as follows: pronation 15-50 degrees (mean, 28.7 degrees) and supination 10-25 degrees (mean, 16.5 degrees). The histopathological examination revealed that there were 5 cases of stage I, 17 of stage II, and 9 of stage III. RESULTS: All patients achieved primary healing of incision and were followed up 1-9 years with an average of 4.5 years. The X-ray films showed that bone healing time was 12-16 weeks with an average of 13 weeks. No tumors recurrence was observed. The ROM of wrist joint was as follows at 1 year after operation: extension 20-50 degrees (mean, 29.0 degrees), flexion 30-50 degrees (mean, 35.0 degrees), radial inclination 10-20 degrees (mean, 16.5 degrees), and ulnar inclination 20-25 degrees (mean, 23.5 degrees). The ROM of forearm was as follows: pronation 40-90 degrees (mean, 68.3 degrees) and supination 30-80 degrees (mean, 59.6 degrees). There were significant differences in the ROM between before operation and after operation (P < 0.05). According to the Krimmer et al wrist score, the results were excellent in 17 cases, good in 12, and fair in 2. CONCLUSION: Wrist joint reconstruction with vascularized fibular head graft can restore function of wrist joint. The operation is proved to be safe and effective in treating distal radius giant cell tumor.

[Diagnosis and choice of surgical methods on traumatic temporomandibular joint ankylosis].

PURPOSE: The purpose of this study was to explore the diagnosis and operative methods for traumatic temporomandibular joint ankylosis. METHODS: Fourteen patients (8 females, 6 males; aged from 7 to 23 years, median age 17.5 years) with traumatic TMJ ankylosis of 1 to 15 years' duration, with a maximal mouth opening(MMO) from 0 to 1.0cm(average:0.46cm) preoperatively were included in this study. They were further divided into two types(I and II) based on false articulation medial to the analysis as shown by coronal computed tomography (CT) coronoid scan. 7 TMJs were treated with medial arthroplasty(MA), in which the lateral fusional bone was removed and the medial false articulation was retained for type I, 6 TMJs were treated with natural positional arthroplasty(NPA) in which the bony fusion between the condyle and glenoid fossa was separated and shaved, the disc was retained and repositioned for type II. Follow-up was carried out and the surgical effect was assessed based on clinical and imaging findings. Student's t test was conducted for comparison with SPSS15.0 software package. RESULTS: There was 9 TMJs from 7 patients for type I and 5 TMJs from 5 patients for type II as shown by CT. The mean postoperative MMO was 3.12cm, and the condylar shape was smooth on CT scan. There was significant difference between preoperative and postoperative MMO, P<0.01. There was no significant difference between MA and NPA, P>0.05. CONCLUSIONS: It is concluded that type I is more common in traumatic TMJ ankylosis than type II. The surgical effect of MA and NPA is good, with less injury, and is worthy of wide application.

Reconstruction of large limb bone defects with a double-barrel free vascularized fibular graft.

BACKGROUND: The use of a free, vascularized fibular graft is an important technique for the reconstruction of large defects in long bones. The technique has many advantages in strong, tubular bones; a more reliable vascular anatomy with a large vascular diameter and long pedicle is used, minimizing donor-site morbidity. Due to limitations in both fibular anatomy and mechanics, they cannot effectively be used to treat large limb bone defects due to their volume and strength. METHODS: From 1990 to 2001, 16 clinical cases of large bone defects were treated using vascularized double-barrel fibular grafts. Patients were evaluated for an average of 10 months after surgery. RESULTS: All the patients achieved bony union; the average bone union took 10 months post surgery, and no stress fractures occurred. Compared with single fibular grafts, the vascularized double-barrel fibular grafts greatly facilitate bony union and are associated with fewer complications, suggesting that the vascularized double-barrel fibular graft is a valuable procedure for the correction of large bone defects in large, long bones in addition to enhancing bone intensity. CONCLUSIONS: The vascularized double-barrel fibular graft is superior to the single fibular graft in stimulating osteogenous activity and biological mechanics for the correction of very large bone defects in large, long bones. Free vascularized folded double-barrel fibular grafts can not only fill up large bone defects, but also improve the intensity margin. Therefore, this study also widens its application and enlarges the treatment targets. However, in the case of bone deformability, special attention should be paid to bone fixation and protection of donor and recipient sites.

[Study on distribution and drainage of lymphatic vessels of tongue].

OBJECTIVE: To investigate the distribution and drainage of lymphatic vessels of tongue, and to provide anatomical evidence for treatment of tongue cancer. METHODS: Indirect lymphatic injection was employed, combined with clearing method with winter green oil and corrosive cast technique, to study the distribution of lymphatic vessels of tongue. Anatomical methods were used to detect the sentinel lymph nodes in different region of tongue. RESULTS: The lymphatic vessels of dorsal mucosa composed of lymphocapillary vessels and anstomosing side branches were present by superficial and deep capillary networks. The distribution of lymphatic networks extend from tip to base and from one board to another, and was not influenced by the sulcus tenninalis and median lingual sulcus. Lymphatic vessels in the muscular portion communicated with lymphocapillary network of dorsal and ventral mucosa, which made the lymphatic vessels of tongue to be an integrity network structure. These characters of distribution influenced the lymphatic drainage of tongue. The results showed principal sentinel lymph nodes (SLNs) for anterior part of tongue were submental lymph nodes, submandibular lymph nodes and juguloomohyoid lymph nodes, for lateral part and middle part of tongue were submandibular lymph nodes, jugulodigastric lymph nodes and thyroid lymph nodes, and for root part of tongue were jugulodigastric lymph nodes. SLNs for every injection region were all presented at bilatral neck, but the frequency of stained SLNs at homolateral neck was more than that at contralateral neck. CONCLUSION: The lymphatic vessels of tongue arranged like a network, which made the lymphatic drainage at various ways and made the distribution of sentinel lymph nodes to be bilateral and dispersive.