Background Triple-negative breast cancer (TNBC) is the most lethal subtype of breast cancer (BC). The circRNA-miRNA‒mRNA axis is a promising biomarker for the early diagnosis and prognosis of BC. However, the critical circRNA mediators involved in TNBC progression and the underlying regulatory mechanism involved remain largely unclear. Methods In this study, we carried out a circRNA microarray analysis of 6 TNBC patients and performed a gene ontology (GO) analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was used to characterize important circRNAs involved in TNBC progression. The interaction between circRNAs and miRNAs was determined by dual luciferase and RNA immunoprecipitation (RIP) assays. Moreover, Transwell, wound healing and Cell Counting Kit-8 (CCK8) assays were performed with altered circRNA or miRNA expression in MDA-MB-231 and BT-549 cells to investigate the roles of these genes in cell invasion, migration and proliferation. Results A total of 78 circRNAs were differentially expressed in TNBC tissues, and the hsa_circ_0045881 level was significantly decreased in TNBC tissues and cells. Lentivirus-mediated hsa_circ_0045881 overexpression in MDA-MB-231 and BT-549 cells significantly reduced cell invasion and migration capacity. Additionally, hsa_circ_0045881 interacted with miR-214-3p in MDA-MB-231 cells. miR-214-3p mimics in MDA-MB-231 and BT-549 cells significantly enhanced cell invasion, migration and proliferation, but the other combinations of inhibitors had opposite effects on cell activity. Conclusions Our data indicated that the circRNA has_circ_0045881 plays key roles in TNBC progression and that hsa_circ_0045881 might act as a sponge for miR-214-3p to modulate its levels in TNBC cells, thereby regulating cell invasion, metastasis and proliferation. hsa_circ_004588 might be a potential prognostic marker and therapeutic target for TNBC.

Background: Testis-specific protein Y-encoded-like 2 (TSPYL2) has been identified as a tumor suppressor on the X chromosome whose hypomethylation is regarded as a biomarker of uterine leiomyomas. Its methylation status in prostate cancer (PCa) re-mains unclear. The purpose of this study was to investigate the methylation status of TSPYL2 in PCa and its role in the migration and proliferation of PCa cells.Methods: Following the required transfection with short hairpin RNA-methyltransferase-like 3 (shRNA-METTL3) and TSPYL2 overexpression plasmid, both quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot assays were adopted to measure the expressions of METTL3 and TSPYL2. The content of N6-methyladenosine (m6A) was measured through RNA immunoprecipitation (RIP). Whether TSPYL2 RNA was methylated in PCa cells was validated by RIP-qPCR. The level of TSPYL2 RNA methylation was determined through Methylated (Me) RIP-qPCR. Scratch and colony formation assays were separately adopted to evaluate migration and proliferation, respectively.Results: TSPYL2 expression was down-regulated (p < 0.001) while METTL3 expression was up-regulated (p < 0.001) in PCa cells (LNCaP and PC3). The expression level of TSPYL2 was down-regulated in prostate adenocarcinoma (PRAD) tumor samples (p < 0.05). m6A content was high in LNCaP and PC3 cells (p < 0.01). TSPYL2 possessed METTL3-related methylation in PCa cells. METTL3 positively regulated TSPYL2 methylation in PCa cells. Both shRNA-METTL3 and TSPYL2 overexpressions decreased the migratory and proliferative abilities of LNCaP and PC3 cells (p < 0.001).Conclusions: TSPYL2-mediated RNA hypomethylation suppresses migration and proliferation of PCa cells.

Background: Bladder cancer (BC) is a highly heterogeneous stem cell disease. Cancer stem cells (CSCs) are the drivers of tumor growth and recurrence, with the ability to self-renew, metastasize, and resist chemotherapy. However, the specific molecular mechanisms of CSCs driving BC recurrence and progression remains unclear.Objective: To explore the underlying molecular mechanism of CSCs driving BC recurrence and progression.Methods: Tumor xenograft model in vivo was established after 4-6-week-old male nude mice were subcutaneously injected with 5×106 of T24 and 5637 cells in 0.1 mL 50% Matrigel. Pearson correlation analysis analyzed the correlation between miR-582-5p and CD81, and which was furtherly verified by dual-luciferase reporter gene assay. Sphere formation assay, flow cytometry, immunohistochemistry (IHC), qRT-PCR and Western blot were carried to examine sphere formation, ALDHhigh populations, the level of genes and proteins. Multivariate analysis was carried to explore the factors associated with recurrence free survival of BC patients.Results: MiR-582-5p was down-regulated in patients with BC, and miR-582-5p overexpression negatively correlated with BC stemness. Mechanically, miR-582-5p negatively targeted to CD81. Functionally, miR-582-5p overexpression inhibited BC stemness and recurrence via targeting CD81.Conclusion: Our study illustrated that miR-582-5p inhibited cell stemness and recurrence via targeting CD81 in BC. Our findings illustrated the specific molecular mechanism of miR-582-5p inhibiting BC progression. MiR-582-5p may serve as the novel biomarker for BC clinical therapeutics and prognosis.

Smoking and Candida albicans (C. albicans) infection are risk factors for many oral diseases. Several studies have reported a close relationship between smoking and the occurrence of C. albicans infection. However, the exact underlying mechanism of this relationship remains unclear. We established a rat infection model and a C. albicans-Leuk1 epithelial cell co-culture model with and without smoke exposure to investigate the mechanism by which smoking contributes to C. albicans infection. Oral mucosa samples from healthy individuals and patients with oral leucoplakia were also analysed according to their smoking status. Our results indicated that smoking induced oxidative stress and redox dysfunction in the oral mucosa. Smoking-induced Nrf2 negatively regulated the NLRP3 inflammasome, impaired the oral mucosal defence response and increased the oral mucosa susceptibility to C. albicans. The results suggest that the Nrf2 pathway could be involved in the pathogenesis of oral diseases by mediating an antioxidative response to cigarette smoke exposure and suppressing host immunity against C. albicans.

OBJECTIVE This study sought to investigate the effect of miR-5191 on proliferation, invasion and metastasis in salivary adenoid cystic carcinoma (SACC). MATERIALS AND METHODS The differential expression level of miR-5191 between 5 primary tumor and adjacent non-neoplastic samples, and in two SACC cell lines was detected by quantitative real-time PCR. Cell proliferation, invasion and migration were performed, followed by luciferase reporter assay and western analysis. The effect of miR-5191 on cell proliferation and apoptosis was evaluated by cell growth and apoptosis assay. The function of miR-5191 in SACC tumorigenesis and metastasis in vivo was investigated by nude mice experiment. The associations between miR-5191/Notch-2 expression and clinicopathological features were analyzed. RESULTS miR-5191 was down-regulated in primary tumor tissues and SACC-LM cells. By targeting Notch-2, miR-5191 expression level affected the migration, invasion and proliferation of SACC cells. Over-expression of miR-5191 inhibited the expression levels of Notch-2, followed by the decreased expression of c-Myc, Bcl-2, Hes-1, Hey-1 and Cyclin D1. In vivo, miR-5191 over-expression suppressed the SACC tumorigenesis and pulmonary metastasis in mice. In SACC patients, higher expression of miR-5191 was related to better prognoses and lower possibility of metastasis. CONCLUSIONS miR-5191 acts as a tumor suppressor in SACC by targeting Notch-2.

KIAA0101, previously identified as PCNA-associated factor, is overexpressed among almost majority of human cancers and has emerged as an important regulator of cancer progression; however, its function in human nasopharyngeal carcinoma (NPC) remain unknown.Integrated bioinformatics approaches were employed to determine the KIAA0101 expressions in the NPC samples.Lentiviral vectors carrying KIAA0101 shRNA were constructed and stable transfected cells were validated by qRT-PCR and western blot.Cellular functions were then evaluated by MTT, colony formation, Brdu staining, and flow cytometry.Mechanistic studies were systematically investigated by UCSC Genome Browser, GEO, UALCAN, QIAGEN, PROMO and JASPAR, ChIP, and the cBioPortal, et al.The results showed that KIAA0101 ranked top overexpressed gene lists in GSE6631 dataset.KIAA0101 was highly expressed in NPC tissues and cell lines.Furthermore, knockdown of KIAA0101 significantly inhibited cell proliferation and DNA replication, promoted apoptosis and cell cycle arrest in vitro.Meanwhile, the mechanistic study revealed that MAP kinase phosphorylation-dependent activation of ELK1 may enhance neighbor gene expressions of KIAA0101 and TRIP4 by binding both promotor regions in the NPC cells.Taken together, our findings indicate that overexpression of KIAA0101 activated by MAP kinase phosphorylation-dependent activation of ELK1 may play an important role in NPC progression.

Cigarette smoke (CS) exposure and Candida albicans (C. albicans) infection are epidemiological risk factors for oral diseases, such as oral leukoplakia (OLK). Smoking-induced inflammation and immune modulation are potentially important mechanisms in the development of diseases, although the biological mechanism of how CS exposure impacts host defenses has not been elucidated. The critical components of host defense, NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome and IL-1β, are required for normal immune function in order to efficiently control infection. This paper studies the molecular mechanism of the immune-suppressive effect of CS on the oral mucosa of animal models. Rats were exposed to intraoral CS to simulate active human smoking and/or to C. albicans for 3 months or 6 months, and their ability to control the infection of C. albicans was examined. The CS and C. albicans co-exposed rats showed early stage lesions of OLK and were more susceptible to C. albicans than those in the C. albicans-exposed group. CS caused a reduced expression of the NLRP3 inflammasome and diminished the secretion of IL-1β and IL-18 maturing by the NLRP3 inflammasome, which were stimulated by C. albicans. CS and immune suppression appear to be closely interwoven at multiple levels. This is the first animal model of active smoking through the mouth, and these data demonstrate that CS suppresses the protective immune response to C. albicans in rats through the NLRP3 inflammasome.

OBJECTIVE:To observe the effects of human umbilical cord mesenchymal stem cells (UC-MSCs) on the proliferation and apoptosis of human ovarian cancer cell SKOV3.
 Methods: Transwell co-culture was used to observe the targeted homing effect of UC-MSCs on ovarian cancer cells. MTT assay was used to detect the inhibitory effect of UC-MSCs conditioned medium on SKOV3 proliferation, and Annexin V-FITC/PI double staining was used to detect the apoptotic rate. Real-time PCR was used to detect the expression levels of Ki-67, Bcl-2 and Bax genes-relevant to proliferation and apoptosis of SKOV3 cells.
 Results: UC-MSCs targeted SKOV3 cells in vitro. MTT assay showed that UC-MSCs conditioned medium significantly inhibited the proliferation of SKOV3 cells (P<0.01). Annexin V-FITC/PI double staining showed that the apoptotic rate in the 75% conditioned medium group was significantly higher than that in the control group (P<0.05). Real-time PCR showed that the expression of proliferation-related gene Ki-67 decreased significantly (P<0.01). The apoptosis-related gene Bcl-2 expression was decreased dramatically (P<0.01), and Bax expression was increased significantly (P<0.01).
 Conclusion: UC-MSCs can target ovarian cancer cells in vitro, inhibit the proliferation of SKOV3 cells by regulating the expression of Ki-67, and promote the apoptosis of SKOV3 cells by regulating the expression of Bcl-2 and Bax.

Background: KIAA0101, previously identified as proliferating cell nuclear antigen (PCNA)-associated factor, is overexpressed among almost majority of human cancers and has emerged as an important regulator of tumorigenesis and cancer progression; however, its expression, function and underlying molecular mechanisms in the development of human nasopharyngeal carcinoma (NPC) remain unknown.Methods: Integrated bioinformatics approaches were employed to determine the differentially expressed genes and KIAA0101 expression levels in the NPC samples compared with normal controls by using gene profiling data from GSE6631, GSE12452 and TCGA-HNSC. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the KIAA0101 mRNA expressions in NPC cell lines. PSCSI-GFP lentiviral vectors carrying KIAA0101 short hairpin RNA (shRNA) were constructed and stable transfected cells were then validated by immunofluorescence, qRT-PCR and western blot for further investigating the effect of KIAA0101 on malignant phenotypes of NPC cells. The effect of KIAA0101 on cell proliferation and apoptosis were evaluated by immunofluorescence, MTT, colony formation and flow cytometry. LinkedOmics was also employed to explore the primary biological process and KEGG pathways correlated with KIAA0101. Flow cytometry and Brdu staining were used to investigate the role of KIAA0101 in the process of cell cycle and DNA replication. Mechanistic studies were systematically investigated by UCSC Genome Browser, GEO, UALCAN, transcriptional factors prediction by QIAGEN, PROMO and JASPAR, ChIP, qRT-PCR, and the cBioPortal for Cancer Genomics, et al.Results: The result showed that KIAA0101 ranked top overexpressed gene lists in GSE6631 expression data. KIAA0101 was highly expressed in the NPC or HNSC tissues and NPC cell lines, and higher expression of KIAA0101 was correlated with tumor grade. Furthermore, knockdown of KIAA0101 significantly inhibited NPC cell proliferation and DNA replication, promoted apoptosis and cell cycle arrest in vitro. Meanwhile, the mechanistic study revealed that MAP kinase phosphorylation-dependent activation of the transcription factor ELK1 enhanced neighbor gene expressions of KIAA0101 and TRIP4 by binding both promotor regions in the NPC cells.Conclusions: Taken together, our findings indicate that overexpression of KIAA0101 regulated by MAP kinase phosphorylation-dependent activation of ELK1 plays an important role in NPC development and progression. Targeting KIAA0101 or ELK1 may provide the promising therapeutic potential for NPC treatment.Funding Statement: This work was supported by the Natural Science Foundation of Jiangsu Province (No. BK20150556), Six Talent Peaks Project in Jiangsu Province (No. YY-076), Project funded by China Postdoctoral Science Foundation (No. 2016M603065, No. 2018T111164) and China Jiangsu Planned Projects for Postdoctoral Research Funds (No. 1601159B).Declaration of Interests: The authors declare that they have no competing interests.Ethics Approval Statement: The study protocol was approved by the Ethics Committee of Medical school of Nanjing University. Data collections and processions were performed according to policies of GEO and TCGA project.

Background T lymphocytes play an important role in contact hypersensitivity. This study aims to explore the immunosuppressive activity of SBF-1, an analog of saponin OSW-1, against T lymphocytes in vitro and in vivo. Methods Proliferation of T lymphocytes from lymph nodes of mice was determined by MTT assay. Flow cytometry analysis was performed to assess T cell activation and apoptosis. Levels of cytokines were determined by PCR and ELISA. BALB/c mice were sensitized and challenged with picryl chloride and thickness of left and right ears were measured. Results SBF-1 effectively inhibited T lymphocytes proliferation induced by concanavalin A (Con A) or anti-CD3 plus anti-CD28 at a very low dose (10 nM) but exhibited little toxicity in non-activated T lymphocytes at concentrations up to 10 μM. In addition, SBF-1 inhibited the expression of CD25 and CD69, as well as he phosphorylation of AKT in Con A-activated T cells. SBF-1 also induced apoptosis of activated T cells. In addition, SBF-1 also downregulated the induction of the T cell cytokines, IL-2 and IFN-γ in a dose-dependent manner. Furthermore, SBF-1 significantly suppressed ear swelling and inflammation in a mouse model of picryl chloride-induced contact hypersensitivity. Conclusions Our findings suggest that SBF-1 has an unique immunosuppressive activity both in vitro and in vivo mainly through inhibiting T cell proliferation and activation. Its mechanism appears to be related to the blockage of AKT signaling pathway.

Objective: The aim of this study was to investigate the effects of Candida albicans on the production of defense effector molecules by human oral mucosal epithelial cells in vitro. Design: Immortalized human oral mucosal epithelial (Leuk-1) cells and C. albicans strain 5314 were cocultured at different cell-to-C. albicans ratios. The viability of Leuk-1 cells was determined by MTT and RTCA measurements. The secretory levels of multiple defense effector molecules were determined by Enzyme-linked immunosorbent assay (ELISA). Results: Our results indicated that C. albicans significantly decreased the secretion of IgG, cystatin C, lactoferrin, and TGF-beta 1 in a dose-dependent manner and remarkably reduced the production of IgA independent of the cell to-C. albicans ratio. However, C. albicans clearly enhanced the secretion of IgM, galectin-3, P-selectin, granzyme B and perforin. Conclusion: These results suggest that C. albicans may exert a regulatory role in the defense response of oral mucosal epithelial cells by altering secretory levels of defense effector molecules.

The aim of this study was to investigate the clinical outcomes of orbital blowout fracture repair by using the three-dimensional (3D) printing-assisted fabrication of individual titanium mesh. Clinical and radiologic data were analyzed for 12 patients with orbital floor and/or medial wall fractures. Lower eyelid incision was used to expose the fractures. Preoperative computed tomographic data were input into an imaging software to rebuild a 3D orbit and mirror the unaffected side into the affected side to replace the demolished orbit. A resin model of the reshaped orbit was generated and used to develop an individual titanium mesh for repairing the fractured orbital. The surgical results were assessed by value of enophthalmos and a comparison of preoperative and postoperative orbital volume difference. All patients had a successful treatment outcome without any complications. Clinical significant enophthalmos were not observed after treatment, and diplopia were solved within 2 weeks postoperative. No extraocular muscle limitation was observed. Postoperative computed tomography scans demonstrated appropriate positioning of titanium mesh and there was no implant displacement. The postoperative orbital volume and enophthalmos difference between the 2 eyes decreased significantly than preoperative (P< 0.001). Three-dimensional printing-assisted fabrication of individual titanium mesh is appropriate for use in orbital blowout fracture.

Objective: This study aimed to investigate the effects of cigarette smoke (extract) on autophagy and apoptosis in oral mucosa epithelial cells. Methods: The effects of cigarette smoke extract (CSE) on autophagy and apoptosis in oral epithelial cells were studied in vivo and in vitro. Leuk-1 cells were administered cigarette smoke extract or chloroquine (CQ) and rapamycin (RAPA) at different concentrations. Immunoblotting, immunofluorescence, Western blotting and flow cytometry were used to detect autophagy-related protein and apoptosis levels, screen the optimal concentration and stimulation time, and verify the effect of CSE stimulation on autophagy and apoptosis in leuk-1 cells. Meanwhile, autophagy expression in epithelial cells from the local oral tissues of mice who had smoked for 5 months was detected. Results: Under CS stimulation, LC3-II and Beclin-1, the key proteins of leuk-1 autophagy, were upregulated in a concentration- and time-dependent manner. In addition, CS significantly upregulated the expression of Cleaved caspase-3 (C-casp3), a protein involved in apoptosis. However, under stimulation with CQ, autophagy in leuk-1 cells was inhibited and the level of C-casp3 and the apoptosis rate were increased. The autophagy activator RAPA significantly reduced the level of C-casp3 and apoptosis rate in leuk-1 cells. Conclusion: The results of this study indicate that CS can simultaneously activate autophagy and apoptosis in mouse and human oral epithelial cells, that autophagy inhibition can aggravate the CSE-triggered apoptosis of oral epithelial cells, and that autophagy induction can inhibit the CSE-triggered apoptosis of oral epithelial cells. Autophagy is suggested to play a protective role in the CSE-induced apoptosis of oral epithelial cells. Further studies are needed to explore the concrete mechanisms underlying the regulatory effects of CS-induced apoptosis and to gain in-depth insight into the complex interactions between apoptosis and autophagy.

Odontoblasts forms dentin as the main hard tissue of the tooth. Autophagy is a highly conserved homeostasis involved in odontoblast differentiation. DSPP (Dentin sialophosphoprotein) and Cbfα1 (Core binding factor α1) are typical factors in odontoblasts of the dentin-pulp complex. However, the relationship between Cbfα1 and DSPP as well as the role of Cbfα1 involved in the autophagy during odontoblast differentiation are poorly understood. In this study, we found the stimulation with FGF2 in MDPC-23 cells resulted in the suppression of autophagy and promotion of cell differentiation. We demonstrated that both mRNA and protein levels of Cbfα1 and DSPP were elevated upon FGF2 stimulation coupled with the inhibition of autophagy in MDPC-23 cells. Further results revealed that overdose of Cbfα1 increased the expression of DSPP, and also suppressed autophagy in MDPC-23 cells. Using luciferase reporter assay and EMSA, we illustrated that Cbfα1 upregulated DSPP expression through binding to the specific sites 5′-TACCTCA (-3950 bp to -3944 bp) and 5′-ACCACA (-3106 bp to -3101 bp) in DSPP promoter. Furthermore, ChIP assay results revealed that FGF2 treatment enhanced this binding of Cbfα1 to DSPP promoter. Signaling pathway inhibitor screening further revealed Cbfα1 suppressed autophagy involved in Wnt I signaling. Moreover, knockdown of Cbfα1 using siRNA silencing approach reversed the FGF2-mediated suppression of autophagy in MDPC-23 cells. Thus, these findings illustrate a novel regulatory mechanism by which Cbfα1 transcriptionally enhances DSPP expression, and also provide an insight into the inhibitory action of Cbfα1 in autophagy thereby facilitating odontoblast differentiation upon FGF2 stimulation.

Objective: This study aimed to study the roles of Muramyl Dipeptide (MDP) which is an agonist of NOD2 in Dendritic Cells (DCs) exposed to Porphyromonas gingivalis lipopolysaccharide ( P. gingivalis - LPS) on maturation and antigen-presenting functions of DCs to provide experimental evidences to explore the possible mechanism of DCs in periodontitis. Methods: Flow cytometry was used to detect CD11c, MHC-II, CD80, CD86, and CD40 expression on DCs and ELISA was used to detect IL-12, IFN-γ, IL-10, and IL-13 secreted by DCs which were stimulated by MDP and P. gingivalis -LPS, respectively or in synergism. RT-PCR analysis was used to detect NOD2, TLR2, and TLR4 mRNA expression in DCs stimulated by MDP and P. gingivalis -LPS, respectively or in synergism. CCK8 was used to assess CD4+ T cells proliferation after co-cultured with DCs stimulated by MDP and P. gingivalis -LPS, respectively or in synergism and ELISA was used to detect IL-2, IFN-γ, IL-10 and IL-13 secreted by these T cells. Results: MDP had weak ability to stimulate DCs maturation but MDP could promote DCs maturation stimulated by P. gingivalis -LPS. MDP was NOD2 agonist to DCs and P. gingivalis - LPS was TLR2 but not TLR4 agonist to DCs. MDP could facilitate TLR2 mRNA expression in DCs exposed to P. gingivalis - LPS. The ability of MDP to promote DCs secreting cytokines was far below P. gingivalis- LPS but MDP could promote the functions of Th2 cell-promoting DCs induced by P. gingivalis -LPS. MDP could promote CD4+T cells proliferation primed by DCs exposed to P. gingivalis -LPS and elevate the ability of DCs exposed to P. gingivalis -LPS to prime Th0 cells to Th2 cells. Conclusion: Intracellular NOD2 in DCs could be activated by MDP and this activation could promote maturation and the ability to prime Th0 cells to Th2 cells of DCs exposed to P. gingivalis -LPS.

Frequent local recurrence and metastasis are generally involved in human liposarcoma, but the management is a challenge. There is an urgent need for improved effective therapy. In the present study, we reported that SBF-1, a steroidal glycoside, inhibited the growth of cultured highly malignant human liposarcoma SW872-S cells in vitro and in vivo. SBF-1 down-regulated the phosphorylation of protein kinase B (AKT) and thus reduced cell adhesion to fibronectin and laminin. Then we found that SBF-1 inhibited the expression of oxysterol binding protein (OSBP) in SW872-S cells, indicating that OSBP may be involved in malignant liposarcoma cell survival. Cancer cell growth and AKT phosphorylation were inhibited significantly upon knockdown of OSBP in SW872-S cells in vitro. Taken together, these results suggest that SBF-1 causes an apparent loss of OSBP function in SW872-S cells, resulting in growth inhibition. Based on our findings, OSBP serves as a potential therapeutic target for human liposarcoma.

This study intended to investigate the role of microRNA-27b (miR-27b) in proliferation of oral squamous cell carcinoma (OSCC) cells and to explore the potential molecular mechanism. Cell proliferation was detected by MTT assay. The expression levels of miR-27b, Frizzled7 (FZD7), cyclin D1 and c-myc were detected by quantitative real time polymerase chain reaction (qRT-PCR). The protein expression level of FZD7 was detected by western blot analysis. The relationship between miR-27b and FZD7, and the activity of Wnt signaling pathway were determined using luciferase reporter assay. The miR-27b expression in OSCC cell lines was significantly decreased compared with control. Overexpression of miR-27b remarkably inhibited OSCC cell proliferation. Additionally, miR-27b could target and inhibit FZD7 expression and decrease the activity of Wnt signaling pathway.miR-27b could inhibit OSCC cell proliferation through inhibiting FZD7 and FZD7-mediated Wnt signaling pathway.

PDGF-BB/PDGFR-ββ signaling plays very crucial roles in the process of many diseases such as liver fibrosis. However, drug candidates with selective affinities for PDGF-B/PDGFR-β remain deficient. Here, we identified a natural cyclopeptide termed destruxin A5 that effectively inhibits PDGF-BB-induced PDGFR-β signaling. Interestingly and importantly, the inhibitory mechanism is distinct from the mechanism of tyrosine kinase inhibitors because destruxin A5 does not have the ability to bind to the ATP-binding pocket of PDGFR-β. Using Biacore T200 technology, thermal shift technology, microscale thermophoresis technology and computational analysis, we confirmed that destruxin A5 selectively targets the PDGF-B/PDGFR-β interaction interface to block this signaling. Additionally, the inhibitory effect of destruxin A5 on PDGF-BB/PDGFR-ββ signaling was verified using in vitro, ex vivo and in vivo models, in which the extent of liver fibrosis was effectively alleviated by destruxin A5. In summary, destruxin A5 may represent an efficacious and more selective inhibitor of PDGF-BB/PDGFR-ββ signaling.

PURPOSE:To investigate the effect of growth factor receptor-bound 7 (Grb7) on oral squamous cell carcinoma growth and tumor xenografts.METHODS:Cal27 and hNOK cells were cultivated, real-time PCR and Western blotting were used to detect the expression of Grb7 in hNOK and Cal27. Cal27 was transfected with Grb7 siRNA for 48 h, cell proliferation was assayed using MTT. Flow cytometry was used to determine the cell cycle and apoptosis. Western blot was used to evaluate the expression of caspase3, Bax, bcl-2, Cyclin D1, Rb, E2F1, ERK and FOXM1. Grb7 siRNA and negative control were designed and injected subcutaneously into the mice, tumor volume and weight were measured. Statistical analysis was performed using SPSS 11.0 software package.RESULTS:Grb7 was highly expressed in Cal27 compared with hNOK. Depletion of Grb7 significantly inhibited cell proliferation, blocked G1/S phase transition, promoted cell apoptosis. Knockdown of Grb7 suppressed the expression of Cyclin D1 and Rb, upregulated E2F1 expression. Moreover, c-caspase 3 and Bax was also reduced after inhibition of Grb7. ERK/FOXM1 signaling pathway was also inhibited by Grb7. In addition, the volume and weight of tumor xenografts were reduced by siGrb7.CONCLUSIONS:Depletion of Grb7 inhibits cell proliferation, promotes apoptosis and reduces tumor xenografts through ERK/FOXM1 pathway.