Crouzon syndrome, a rare genetic disorder characterized by premature closure of coronal sutures, results in skull and facial deformities along with abnormal brain and ocular development.Here, we report a case of a 27-year-old ethnic han male patient who presented with complex binocular strabismus secondary to Crouzon syndrome. At the time of surgery, extraocular muscles were found to be fibrotic and results of the pathological examination revealed degeneration of muscle fibers, which were replaced by adipose tissue. The entire exome sequencing DNA testing indicated that the patient and his father possessed the fibroblast growth factor receptor 2 (FGFR2) gene c.G812T:p.G271V heterozygous mutation. Binocular strabismus corrective surgery was performed in this patient with a satisfactory outcome.This case demonstrates that Crouzon syndrome patients can show an FGFR2 gene c.G812T:p.G271V mutation and display clinical symptoms such as extraocular muscle fibrosis, exotropia, exophthalmos, and a pointed head deformity.
Abstract Background P53 pathway inactivation plays an important role in the process of breast cancer tumourigenesis. Post-translational protein modification abnormalities have been confirmed to be an important mechanism underlying the inactivation of p53. Numerous deubiquitinating enzymes are aberrantly expressed in breast cancer, and a few deubiquitination enzymes are capable of deubiquitinating and stabilizing p53. Here, we report that OTUD3 is a deubiquitylase of p53 in breast carcinoma. Methods The correlation between the mRNA expression of OTUD3, TP53 and PTEN and the prognosis of BC was assessed with the Kaplan-Meier Plotter tool. OTUD3 protein expression in breast carcinoma was examined by immunohistochemistry and western blotting. The relationship among OTUD3, p53, and p21 proteins was analysed. Half-life analysis and ubiquitylation assay were performed to elucidate the molecular mechanism by which OTUD3 stabilizes p53. The interaction between OTUD3 and p53 in BC cells was verified by a co-immunoprecipitation assay and GST pulldown experiments. MTS proliferation detection, an apoptosis detection kit and colony formation asssy were used to investigate the functional effects of OTUD3 on breast cancer cells. Results OTUD3 downregulation is correlated with a poor prognosis in BC patients. OTUD3 expression is decreased in breast cancer tissues and independent of the histological grade.OTUD3 also inhibits cell proliferation and clone formation and increases the sensitivity of BC cells to apoptosis induced by chemotherapy drugs. A reduction in OTUD3 expression concomitant with decreased p53 abundance is correlated with human breast cancer progression. The ectopic expression of wild-type OTUD3, but not its catalytically inactive mutant, stabilizes and activates p53. Mechanistically, OTUD3 interacts directly with p53 through the amino-terminal OTU region. Finally, OTUD3 protects p53 from Mdm2-mediated ubiquitination and degradation, enabling the deubiquitination of p53 in BC cells. Conclusions In summary, we establish that OTUD3 is a potential therapeutic target for restoring p53 function in breast cancer cells and suggest that the OTUD3-p53 signalling axis plays a critical role in tumour suppression.
Background and Objective: The circulating immune cells mobilized during liver transplantation may play important roles in acute phase graft injury and late phase tumor recurrence. However, the precise mechanism of circulating immune cell mobilization during liver transplantation has not been explored. In the current study, we aim to investigate the impact of acute-phase small-for-size graft injury on mobilization of circulating regulating B cells (Bregs) in the patients with hepatocellular carcinoma (HCC) after liver transplantation and to explore the underlying molecular mechanism therein. Methods: From May 2000 to November 2009, 115 HCC recipients were included in the current analysis. The intragraft gene expression profile together with Bregs intragraft infiltration were compared between the recipients implanted with liver grafts greater (Group 1) and less than 60% (Group 2) of standard liver weight (SLW) by real-time RT-PCR and immunostaining, respectively. Circulating Bregs (CD19+CD24hiCD38hi) were detected by FACS analysis at different time points after liver transplantation. Clinical-pathological data including the incidence of tumor recurrence and metastasis were compared between the two groups. The direct roles of TLR4, CXCL10 and CXCR3 on circulating Bregs mobilization were further investigated in TLR4-/-, CXCL10-/- and CXCR3-/- mice models, respectively. The association of intragraft Bregs infiltration and tumor invasiveness were also examined in the rat liver transplantation for liver cancer model. The role of Bregs on liver tumor growth and invasiveness were further studied in a series of in vitro and in vivo functional experiments. Results: The patients were grouped into Group 1 (>= 60% SLW, n=37) and Group 2 (< 60% SLW, n=78). The numbers of patients beyond Milan criteria [15/37(40.5%) vs 29/49(59.2%), p=0.838] or UCSF criteria [9/37(24.3%) vs 19/60(31.7%), p=1] were similar in the two groups. Much more patients in Group 2 developed tumor recurrence and lung metastasis [19/78(24.4%) vs 3/37(8%), p=0.04]. The level of circulating Bregs was significantly higher in Group 2 (Week1: 7.02 vs 1.31/10ˆ5PBMC, p=0.03; month3: 5.7 vs 1.3/10ˆ5PBMC, p=0.03). There was more intragraft Bregs infiltration in group 2 indicated by CD20/IL10 staining. Intragraft gene expression of TLR4, CXCL10 and CXCR3 were significantly higher in Group 2 at early phase after transplantation. In rat liver transplantation model, there was more Bregs infiltration at early phase after transplantation correlating with late phase invasive tumor growth. Levels of circulating Bregs were significantly lower in the mice model with major hepatectomy and hepatic I/R injury using TLR4-/-, CXCL10-/- and CXCR3-/- mice, respectively. In the in vitro co-culture system, Bregs enhanced liver cancer cell proliferation and migration. In the scid mice liver cancer model, Bregs injection significantly promoted tumor growth. Conclusion: A significantly higher population of circulating Bregs, which are mobilized by small-for-size graft injury, may lead to a higher incidence of tumor recurrence and metastasis after LDLT. TLR4/CXCL10/CXCR3 signaling may play important roles on Bregs mobilization.
Objective To evaluate the effects of Met inhibitor XL-880 on radiosensitivity of breast cancer cells MDA-MB-231.Methods MDA-MB-231 cell lines were assigned to the following treatment groups:control group,radiation group,XL-880 group and combination group.Cell apoptosis,cell cycle distributions and tumorigenicity were investigated by flow cytometry or clonogenic assay.The expression of apoptosis and cell cycle related proteins (p21,Cyclin B1,Bcl-2,Caspase-3 and PARP),and phosphorylation levels of c-Met were measured by Western blot.Results XL-880 combined with radiation significantly decreased the proliferation activity of MDA-MB-231 cells (P ＜ 0.05).Flow cytometry results showed that the rate of G2/M cell were increased with XL-880 (P ＜ 0.05),and the rate were (17.3 ±1.3) ％,(20.0 ± 4.0) ％,(28.5 ± 3.1) ％,(57.0 ± 3.3) ％,respectively.Annexin V/PI double-staining assay showed that XL-880 obviously induced the apoptosis of MDA-MB-231 cells after radiation (P ＜ 0.05),of which the apoptotic rates were (7.3 ±0.9)％,(14.1 ±0.6)％,(35.5 ±4.4)％,(48.2±5.3)％,respectively.XL-880 downregulated the expressions of Cyclin B1 and anti-apoptosis protein Bcl-2,while promoted the expression of apoptosis related protein cleaved Caspase-3 and PARP.Conclusions XL-880 enhance the radiosensitivity of breast cancer cell MDA-MB-231 by inhibiting Met pathway.
Key words:
Breast neoplasms; Radiotherapy; Proto-oncogene proteins c-met
Objective To investigate the effects of fibroblast growth factor 2 (FGF2) reversing transforming growth factor β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of lung cancer cell A549.Methods Cell scratch and Transwell methods were used to examine the migration and invasion ability of cancer cells.The expression changes of key EMT and MET molecular maker E-cadhefin,Vimentin,N-cadherin,and Snail were measured by using immunofluorescence and Western blotting.Signaling pathways were analyzed by using selective inhibitors LY294002 and PD98059.Results The morphological changes and the epithelial and mesenchymal markers induced by TGF-β1 were reversed by FGF2 to control levels.FGF2 also decreased the migration and invasion ability of A459 cells (50.0 ± 5.5,P ＜ 0.01).Signaling pathways analyzed by selective inhibitors showed that the inhibition of MAPK/ERK kinase pathway could substantially block the reversal effects of FGF2,while the activation of MAPK/ERK kinase pathway resultd in Smad2 dephosphorylation.Conclusion These findings indicate that the TGF-β1-induced EMT is reversed by FGF2 through the MAPK/ERK kinase pathway.
Key words:
Lung carcinoma; Transforming growth factor β1 ; Fibroblast growth factor 2; Epithelial-mesenchymal transition
MicroRNAs (miRNAs) have been identified as negative posttranscriptional regulators of target genes and are involved directly in the pathological processes of tumors, including drug resistance. However, the exact function of miR-520h in breast cancer remains poorly understood. The aim of this study was to investigate the molecular mechanisms of miR-520h in paclitaxel resistance in the MCF-7 breast cancer cell line. Ectopic expression of miR-520h could promote the proliferation of breast cancer cells and inhibit paclitaxel-induced cell apoptosis. Inhibiting the expression of miR-520h could enhance the sensitivity to paclitaxel in paclitaxel-resistant MCF-7/Taxol cells. Furthermore, luciferase reporter assays showed that OTUD3 was a direct target of miR-520h. OTUD3 plays a necessary role in the paclitaxel resistance effect of miR-520h, and cotreatment with a miR-520h inhibitor and OTUD3 overexpression significantly enhanced MCF-7 cell sensitivity to paclitaxel. Moreover, miR-520h substantially inhibited the protein expression of PTEN via OTUD3 and subsequently affected downstream p-AKT pathway activity. In a clinical study, we also found that high miR-520h expression was associated with more aggressive pathological characteristic and poor prognosis. Therefore, our findings showed that miR-520h targeted the OTUD3-PTEN axis to drive paclitaxel resistance, and this miR might be an important potential target for breast cancer treatment.
Background: Breast cancer (BRCA) is a highly heterogeneous malignancy. Current molecular typing, such as PAM50, has certain limitations.Methods: DNA methylation-related genes (METcor) and microRNA (miRNA)-related genes (MIRcor) were screened using The Cancer Genome Atlas-Breast Cancer (TCGA-BRCA) samples. Genes were then integrated, and their mRNA expression profiles were clustered to classify BRCA into three subtypes (named Gao typing), including Immune activation-associated subtype (IAA), Proliferation-associated subtype (PA), and Estrogen-associated subtype (EA).Findings: Gao typing can further stratify PAM50 subtypes, particularly in the poorer prognostic Her2-enriched and TNBC subtypes. By screening differentially expressed genes (DEGs) among subtypes and 37 vital prognostic genes were finally obtained through lasso regression analysis. A prognostic risk factor was constructed based on lasso regression coefficients, and a new prognostic model for BRCA was constructed by combining Gao typing, the prognostic risk factor, and clinicopathological parameters. The prognostic model was found to have a robust survival prediction for BRCA patients.Interpretation: For the first time, we developed a novel molecular classification method (Gao typing) for breast cancer based on the co-regulation of DNA methylation and miRNA on mRNA expression profiles.Funding Information: This work was supported by the Chinese National Natural Science Foundation Projects Grant No.81572587, Key projects of Qingdao science and technology plan Grant No.19-6-1-4-nsh and 20-3- 4-40-nsh, Shandong University Qilu Hospital (Qingdao) Flexible Talent Guidance Project Grant No.QDKY2021RX02.Declaration of Interests: None.Ethics Approval Statement: All patients or relatives informed and consented to the use of their samples, this study was also approved by the Ethics Committee of Qilu Hospital of Shandong University (Qingdao) (KYLL-KSqdql2018011).
Transplant rejection is a major cause of corneal transplantation failure. MicroRNAs (miRNAs) are a family of small RNAs that regulates gene expression in a sequence-specific manner. miRNAs have recently been shown to have important roles in human organ transplantation, but reports of miRNAs directly associated with corneal transplantation rejection remain limited. To investigate the role of miRNAs during corneal allograft rejection, we established a mouse penetrating keratoplasty model and used microarrays to screen for differentially expressed miRNAs. Our results revealed that the expression of miR-122 was significantly decreased in the allogeneic group. Consistent with this result, the expression of cytoplasmic polyadenylation element-binding protein-1 (CPEB1), a direct target of miR-122, was significantly increased. Further analysis demonstrated that miR-122 inhibited inflammatory cytokine-induced apoptosis in corneal keratocytes through the downregulation of its target CPEB1. We also found that increased miR-122 expression significantly reduced the risk of corneal transplantation rejection. Thus, our results indicate that miR-122 is an important miRNA associated with corneal graft rejection and can be used as a therapeutic target for the prevention of immune rejection after keratoplasty.
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