ABSTRACT
Collagen type IV α1 chain (COL4A1) is a collagen protein that acts as a tumor-promoting factor in several types of cancer. However, the role and the potential mechanisms involving COL4A1 in oral squamous cell carcinoma (OSCC) remain unclear. Using reverse transcription-quantitative PCR and western blotting, the expression levels of COL4A1 and (nidogen-1) NID1 in OSCC cells were assessed. Cell Counting Kit-8, EdU staining and colony formation assays were used to evaluate cell proliferation. Cell migration and invasion were assessed using wound healing and Transwell invasion assays, respectively. The expression levels of proteins involved in epithelial-mesenchymal transition (EMT) were assessed using western blotting. In addition, the association between COL4A1 and NID1 was analyzed using TNMplot and the STRING database and verified by co-immunoprecipitation analysis. COL4A1 expression was found to be significantly increased in OSCC cells. Knockdown of COL4A1 expression decreased SCC-4 cell proliferation, migration and invasion, as well as the progression of EMT. In addition, COL4A1 was shown to be significantly positively associated with NID1 in OSCC and to bind to NID1. NID1 overexpression reversed the inhibitory effects of COL4A1 knockdown on cell proliferation, migration and invasion as well as on the progression of EMT in OSCC cells. In summary, the present findings demonstrated that COL4A1 promoted cell proliferation and migration as well as the progression of EMT in OSCC cells by binding to NID1, highlighting a potential avenue for therapeutic management of OSCC.
ABSTRACT
It has been reported that protein arginine methyltransferase 5 (PRMT5) serves a significant role in osteogenic differentiation and inflammatory response. Nevertheless, its role in periodontitis as well as its underlying mechanism remain to be elucidated. The aim of the present study was to explore the role of PRMT5 in periodontitis and whether PRMT5 could reduce liposaccharide (LPS)-induced inflammation of human periodontal ligament stem cells (hPDLSCs) and promote osteogenic differentiation through STAT3/NF-κB signaling. In the current study, the expression levels of PRMT5 were determined in LPS-induced hPDLSCs by reverse transcription-quantitative PCR and western blot analysis. ELISA and western blot analysis were employed to assess the secretion and expression levels of inflammatory factors, respectively. The osteogenic differentiation and mineralization potential of hPDLSCs were evaluated using alkaline phosphatase (ALP) activity assay, Alizarin red staining and western blot analysis. Additionally, western blot analysis was applied to determine the expression levels of the STAT3/NF-κB signaling pathway-related proteins. The results showed that the expression levels of PRMT5 were significantly enhanced in LPS-induced hPDLSCs. Additionally, PRMT5 knockdown reduced the contents of IL-1ß, IL-6, TNF-α, inducible nitric oxide synthase and cyclooxygenase-2. PRMT5 depletion also enhanced ALP activity, improved the mineralization ability and upregulated bone morphogenetic protein 2, osteocalcin and runt-related transcription factor 2 in LPS-induced hPDLSCs. Furthermore, PRMT5 knockdown inhibited inflammation and promoted the osteogenic differentiation of hPDLSCs via blocking the activation of the STAT3/NF-κB signaling pathway. In conclusion, PRMT5 inhibition suppressed LPS-induced inflammation and accelerated osteogenic differentiation in hPDLSCs via regulating STAT3/NF-κB signaling, thus providing a potential targeted therapy for the improvement of periodontitis.
ABSTRACT
OBJECTIVE: To investigate the effect of bradykinin (BK) on cisplatin (DDP)-induced cardiotoxicity at the cellular level and its cytological mechanism. METHODS: The toxic effects of DDP on GP-H1 cells, and the effects of BK on DDP cardiomyocyte survival rate, DDP-induced malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD), reactive oxygen species (ROS), mitochondria membrane potential (MMP) and apoptosis were explored. RESULTS: DDP at different concentrations inhibited GP-H1 cells at 12 h after administration, and the inhibitory effect was more prominent at 24 h after administration and continued until 72 h after administration. The severity of GP-H1 cell damage induced by DDP was reduced by 0.1 µM, 1 µM, and 10 µM BK. After GP-H1 cells were treated with DDP, ROS levels increased and MMP levels decreased, while BK intervention inhibited these effects. At 24 h after DDP treatment, Bax/bcl-2 increased in GP-H1 cells, and the expressions of Caspase-3, p-NF-κB, p-p38 and p-Smad2 decreased. After intervention with BK, it was shown that Bax/Bcl-2 was significantly reduced, and the expressions of Caspase-3, p-NF-κB, p-p38 and p-Smad2 decreased. Bax/Bcl-2 and the expressions of Caspase-3, p-NF-κB, p-p38 and p-Smad2 of GP-H1 cells were basically not affected by BK alone. CONCLUSION: The protective effect of BK on DDP-induced GP-H1 cell damage in guinea pig is related to the activation of PI3K/Akt/NO signaling pathway by BK, which reduces oxidative stress levels in cardiomyocytes and also acts as an anti-apoptotic agent.
ABSTRACT
BACKGROUND: We aimed to evaluate the efficacy of mineral trioxide aggregate (MTA) in the treatment of endodontic disease. METHODS: Oversell, 384 patients with endodontic disease treated in Xuzhou Stomatological Hospital, Xuzhou, China, from June 2015 to June 2017 were selected, and randomly divided into four groups with 96 cases per group. The repair effects of MTA, zinc phosphate cement (ZPC), silver amalgam and light-curing calcium hydroxide (LCH) on the teeth and dental pulp of patients in the four groups were compared. Meanwhile, the ill symptoms of the patients were observed to confirm whether they could be alleviated. Besides, whether the repair effects were related to ages of patients, perforation diameters of diseased teeth and repair materials was discussed. RESULTS: The success rates of MTA group, ZPC group, LCH group and silver amalgam group were 90.6%, 68.7%, 70.8% and 52.1%, respectively. The success rate of MTA group was significantly higher than that of ZPC group, silver amalgam group and LCH group. When the success rates of four groups were compared, the differences were statistically significant (P=0.0072). The patient's age, repair material and perforation diameter were positively correlated with MTA repair effect (P=0.003, P=0.002, P=0.01). The patients' teeth in each group were repaired with different materials, and the reexamination was conducted 4 weeks later. Three patients in the silver amalgam group were found to have gingival swelling. CONCLUSION: The therapeutic effect of MTA was significant in the treatment of endodontic disease, and it is worthy of clinical application.
ABSTRACT
The aim of this study was to investigate and discuss the toxic effect of four kinds of dental restorative materials on fibroblast HGF-1 and their impacts on the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) genes. One hundred and ninety-two patients (240 diseased teeth), who received dental restoration in the Department of Stomatology of Xuzhou Stomatology Hospital from March 2014 to March 2015, were selected and randomly divided into four groups; namely, silver amalgam group, glass-ionomer cement group, nichrome group and novel nano-composite resin group, with 60 teeth in each group. The diseased teeth were restored. The fibroblast HGF-1 was incubated in the water extracts from the four kinds of materials and ordinary cell culture fluid (negative control). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting were used to measure the levels of Bcl-2 and Bax. A flow cytometer was applied to detect cell apoptosis. RT-qPCR results showed that compared with those in the negative control group, the mRNA expression levels of Bcl-2 in the cells of silver amalgam group, glass-ionomer cement group and nichrome group were decreased, while those of Bax were upregulated (P<0.05). The mRNA expression of Bcl-2 in glass-ionomer cement group was the highest among these three groups; the mRNA expression of Bax in nichrome group was the highest of all groups. The western blotting results revealed the same tendency as those of RT-qPCR. The results via the flow cytometer showed that cell apoptosis in nichrome group, silver amalgam group and glass-ionomer cement group was increased significantly (P<0.05) compared with that in the negative control group. The novel nano-composite resin has no obvious toxic effect on cells, and its clinical application effect is better than that of traditional dental restorative materials, which is worthy of application and generalization in clinical practice.
ABSTRACT
OBJECTIVE: To establish an efficient and stable method for protein extraction of Streptococcus mutans. METHODS: The collected bacteria were treated by freeze-thaw and ultrasonic (method 1), ultrasonic (method 2), boiling (method 3), boiling and ultrasonic (method 4), respectively. The index such as state of bacteria broken, concentration of extracted protein and SDS-PAGE of protein were employed to evaluate the effects of above four methods. RESULTS: Beside the method 3, the other three methods could break the bacteria effectively, of which ultrasonic was the key factor. The pattern of SDS-PAGE which treated by method 1, method 2 and method 4 was almost same, but method 1 resulted in the best abundance. There was significantly difference among the four protein concentration extracted by four methods (P < 0.05). All methods exhibited good stability and reproducibility. CONCLUSION: Method of freeze-thaw and ultrasonic resulted in an efficient proteins extraction of Streptococcus mutans which demonstrated good stability and reproducibility and easy to handle.