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@#Objective To investigate the effects of the long non-coding RNA LOXL1 antisense RNA 1 （LOXL1-AS1） on the apoptosis and inflammatory factor expression of human brain microvascular endothelial cells （HBMECs） induced by oxygen-glucose deprivation （OGD）. Methods HBMECs were divided into control group （normal culture）， OGD group （OGD injury）， OGD+si-NC group （transfection with si-NC plus OGD injury）， OGD+si-LOXL1-AS1 group （transfection with si-LOXL1-AS1 plus OGD injury）， OGD+miR-NC group （transfection with miR-NC plus OGD injury）， OGD+miR-761 group （transfection with miR-761 mimic plus OGD injury）， OGD+si-LOXL1-AS1+negative control group （transfection with si-LOXL1-AS1 and anti-miR-NC plus OGD injury）， and OGD+si-LOXL1-AS1+miR-761 inhibitor group （transfection with si-LOXL1-AS1 and miR-761 inhibitor plus OGD injury）. The expression of LOXL1-AS1 and miR-761 was measured by RT-qPCR. Cell viability was measured using cell counting kit-8. Cell apoptosis was determined by flow cytometry. The expression of B-cell lymphoma/leukemia-2 （Bcl-2） protein and Bcl-2-associated X （Bax） protein was measured by Western blotting. The levels of interleukin （IL）-6， IL-1β， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay. Dual luciferase reporter assay was used to detect the complementary binding of LOXL1-AS1 and miR-761. Results Compared with the control group， the OGD group showed significant increases in LOXL1-AS1 expression， the cell apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels and significant decreases in the cell survival rate and Bcl-2 expression （all P<0.05）. After inhibiting LOXL1-AS1， the OGD+si-LOXL1-AS1 group showed significant decreases in LOXL1-AS1 expression， the apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels and significant decreases in the survival rate and Bcl-2 expression， compared with the OGD group and the OGD+si-NC group （all P<0.05）. LOXL1-AS1 targeted the expression of miR-761. After overexpressing miR-761， the OGD+miR-761 group showed significant increases in the survival rate and Bcl-2 expression and significant decreases in the apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels， compared with the OGD+miR-NC group （all P<0.05）. Compared with the OGD+si-LOXL1-AS1+negative control group， the OGD+si-LOXL1-AS1+miR-761 inhibitor group showed significantly decreased survival rate and Bcl-2 expression and significantly increased apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels （all P<0.05）. Conclusion Inhibiting LOXL1-AS1 expression can up-regulate miR-761 to promote the survival of OGD-induced HBMECs and suppress the cells' apoptosis and expression of inflammatory factors.
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Objective @#To observe the dynamic expression of recombinant lysyl oxidase like protein 1 ( LOXL1) in the lysine oxidase family in the liver of C57BL/6 mice infected with Schistosomajaponicum and explore its role in hepatic fibrosis．@*Methods@#Mice were infected subcutaneously with cercariae of S．japonicum，and sacrificed with euthanasia in 6，9 and 12 weeks after infection．The sera and liver tissues were collected．The levels of liver fibrosis in mice was dynamically evaluated by HE and Sirius red staining，and the serum transaminases were detected．The dynamic expression levels of collagen type Ⅰ ( Col1) ，LOXL1 and α-smooth muscle actin( α-SMA) in liver tissues were determined respectively by Western blot and qPCR. Finally，the dynamic levels of soluble and insoluble collagens were detected. @*Results@#The result of HE and Sirius red staining showed that hepatic fibrosis levels increased at 6 weeks，peaked at 9 week，and decreased at 12 week in response to S．japonicum infection．Western blot and q-PCR showed that the expression levels of LOXL1，Col1 α1，Col3 α1 and α-SMA was significantly up regulated and reached maximum at the 9th week in response to S．japonicum infection．Soluble collagen protein levels reached maximum at the 9th week．and decreased at 12 week，however insoluble collagen protein levels continued to increase．@*Conclusion@#There may be a correlation between LOXL1 and fiber cross-linking in the process of hepatic fibrosis in S．japonicum，and it plays a role in promoting hepatic fibrosis.
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Abstract Introduction: Lysyl oxidase-like 4 is an amine oxidase from the lysyl oxidase family that was previously shown to be overexpressed in head and neck cancer and upregulated in response to hypoxia. The possible role of lysyl oxidase-like 4 as a tumor marker in advanced stage larynx cancer was investigated. Objective: To investigate the expression of lysyl Oxidase-Like 4 protein in advanced stage laryngeal cancer and elucidate its possible role as a tumor marker, predictor of treatment response and prognosticator. Methods: Diagnostic specimens of 72 patients treated for stage III-IV laryngeal squamous cell carcinoma were evaluated for lysyl oxidase-like 4 expression by immunohistochemistry. Results: Lysyl oxidase-like 4 expression was correlated with advanced tumor stage (p = 0.041) and better differentiation (p = 0.025) but was independent of tumor diameter (p = 0.456). Response to induction chemotherapy or the need for salvage laryngectomy were not affected by lysyl oxidase-like 4 expression (p = 0.999, p = 0.070 respectively). Increased lysyl oxidase-like 4 expression was associated with better 2 year overall survival in both univariate (p = 0.036) and multivariate analyses (p = 0.014). Conclusion: Lysyl oxidase-like 4 expression emerges with advancing stages, is lost with worsening differentiation, and may have tumor suppressive properties in larynx cancer.
Resumo Introdução: A proteína tipo-lisil oxidase-4 é uma amina oxidase da família lisil oxidase cuja superexpressão em câncer de cabeça e pescoço e up-regulação em resposta à hipóxia foram previamente demonstradas. O possível papel da proteína tipo-lisil oxidase-4 como um marcador tumoral no câncer de laringe em estágio avançado foi investigado. Objetivos: Investigar a expressão da proteína tipo-lisil oxidase-4 no câncer de laringe em estágio avançado e elucidar seu possível papel como marcador tumoral, preditor da resposta ao tratamento e do prognóstico. Método: Amostras diagnósticas de 72 pacientes tratados para carcinoma espinocelular da laringe em estágio III-IV foram avaliadas quanto à expressão da proteína tipo-lisil oxidase-4 por imuno-histoquímica. Resultados: A expressão de proteína tipo-lisil oxidase-4 foi correlacionada com o estágio avançado do tumor (p = 0,041) e melhor diferenciação (p = 0,025), mas foi independente do diâmetro do tumor (p = 0,456). A resposta à quimioterapia de indução ou a necessidade de laringectomia de resgate não foram afetadas pela expressão da proteína tipo-lisil oxidase-4 (p = 0,999, p = 0,070 respectivamente). O aumento da expressão da proteína tipo-lisil oxidase-4 foi associado a melhor sobrevida global de 2 anos nas análises univariada (p = 0,036) e multivariada (p = 0,014). Conclusão: A expressão da proteína tipo-lisil oxidase-4 surge com o avanço dos estágios e desaparece com pioria da diferenciação e pode ter propriedades supressoras de tumor no câncer de laringe.
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Lysyl oxidase like 4 (LOXL4) is one member of the LOX protein family and is a secreted copper-dependent amine oxidase involved in the assembly and maintenance of extracellular matrix (ECM). LOXL4 is up-regulated in human liver cancer, gastric cancer, breast cancer, cervical cancer, head and neck squamous cell carcinoma, esophageal carcinoma and colorectal cancer, but down-regulated in human bladder and lung cancer. It also inhibits tumor growth in bladder and lung cancer, suggesting that L0XL4 has a dual role of promoting or inhibiting tumors in different types of human malignant tumors. L0XL4 in tumor cell exosomes promotes cell matrix adhesion and cell migration by activating the FAK/Src pathway, which is dependent on its amine oxidase activity through a hydrogen peroxide-mediated mechanism. Exosome-mediated L0XL4 can also promote tumor cell proliferation and immune escape by activating the PI3K/Akt signaling pathway. L0XL4 can be transported to macrophages via exosomes in tumor cells, where it further activates the immunosuppression function of cells and the expression of programmed death ligand 1 (PD-L1) via STAT1- and STAT3-mediated signaling pathways. It then will trigger the immunosuppressive function of macrophages and promote the immune escape of tumor cells. In addition, LOXL4 can also exert the tumor suppressive function by activating p53 and inhibiting the Ras/ERK pathway. This paper mainly reviews the structure and the function of LOXL4, and the relationship between LOXL4 and the pathogenesis and development of human malignant tumors. We then further explore the application of LOXL4 in the study of malignant tumors, laying a theoretical foundation for its future utilization in the clinical diagnosis and treatment, and screening of prognostic markers of human malignant tumors.
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Abstract Objectives Diabetes has been strongly associated with periodontal diseases. The periodontal ligament (PDL) has an abundant extracellular matrix (ECM). Lysyl oxidases (LOXs) are closely associated with various diseases caused by abnormal ECM functions, however, the role of LOXs in periodontal diseases induced by diabetes remains unclear. Methodology In this study, 8-week-old Zucker diabetic fatty rats were used to establish a type 2 diabetes mellitus (T2DM) model. After 9 and 16 weeks, hematoxylin and eosin (H&E), Masson's trichrome, and immunohistochemical staining were performed. Results After 9 weeks, loose collagen fibers were found in the interradicular area of the diabetic group, in opposition to the control group. There were no significant differences in LOX expression between the diabetic and control groups (p>0.05). However, after 16 weeks, the diabetic group presented a disordered arrangement of the PDL, showing decreased collagen content and significantly increased lysyl oxidase-like protein 3 (LOXL3) expression when compared with the control group (p<0.05). This suggests that LOXL3 plays a significant role in periodontal histopathological changes in diabetic rats. Conclusion Our study showed elevated LOXL3 expression in the PDL of diabetic rats after 16 weeks, suggesting that LOXL3 may be involved in the occurrence and development of periodontal histopathological changes in diabetic rats. LOXL3 could be further used as an indicator for the early diagnosis of diabetic periodontitis in T2DM patients in clinical settings.
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Objective To investigate the expression and clinical significance of S100A14 and LOXL2 in papillary thyroid carcinoma tissues. Methods Paraffin blocks from 90 cases of thyroid lesion were collected to make tissue microarray. The expression of S100A14 and LOXL2 in 30 cases of nodular goiter (NG), 30 cases of classic papillary thyroid carcinoma (CPTC) and 30 cases of high invasion papillary thyroid carcinoma were detected by immunohistochemistry. The correlation of S100A14 and LOXL2 expression with the clinicopathological characteristics of papillary thyroid carcinoma patients was investigated. Results S100A4 and LOXL2 were highly expressed in papillary thyroid carcinoma. There was a gradually increasing trend from the expression in NG group, CPTC group to high invasive group. The positive rate of S100A14 in papillary thyroid carcinoma with lymph node metastasis was higher than that without lymph node metastasis (P < 0.05). There was significant difference of S100A14 and LOXL2 expression between CPTC group and high invasion group (P < 0.05). S100A14 expression was positively correlated with LOXL2 expression (r=0.332). Conclusion The high expression of S100A14 and LOXL2 may be related to the malignant biological behavior and invasiveness of papillary thyroid carcinoma, and S100A14 may promote lymph node metastasis.
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@#The lysyl oxidase family has five family members which are; Lysyl Oxidase (LOX), Lysyl Oxidase Like-1 (LOXL1), Lysyl Oxidase Like-2 (LOXL2), Lysyl Oxidase Like-3 (LOXL3), and Lysyl Oxidase Like-4 (LOXL4). These are amine oxidases which are copper (Cu) dependent. The main function of these secreted enzymes is covalently crosslinking extracellular collagens and elastins, making the extracellular matrix (ECM) stable. Association with LOX family enzymes has been found in various diseases including tumours, suggesting that it may be involved in the pathogenesis of the lesions. To add to the complexity, some of the LOX family members have been linked with tumour suppression while the other members were associated with tumour promotion, progression and metastasis. Thus, this review will explore further insight into the role of LOX family in tumour formation.
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BACKGROUND/AIMS: Lysyl oxidase-like 2 (LOXL2), a collagen-modifying enzyme, has been implicated in cancer invasiveness and metastasis. METHODS: We evaluated the expression of LOXL2 protein, in addition to carbonic anhydrase IX (CAIX), keratin 19, epithelial cell adhesion molecule, and interleukin 6, in 105 resected hepatocellular carcinomas (HCCs) by immunohistochemistry. RESULTS: LOXL2 positivity was found in 14.3% (15/105) of HCCs, and it was significantly associated with high serum α-fetoprotein levels, poor differentiation, fibrous stroma, portal vein invasion, and advanced TNM stage (p < 0.05 for all). Additionally, LOXL2 positivity was significantly associated with CAIX (p=0.005) and stromal interleukin 6 expression (p=0.001). Survival analysis of 99 HCC patients revealed LOXL2 positivity to be a poor prognostic factor; its prognostic impact appeared in progressed HCCs. Furthermore, LOXL2 positivity was shown to be an independent predictor of overall survival and disease-specific survival (p < 0.05 for all). Interestingly, co-expression of LOXL2 and CAIX was also an independent predictor for overall survival, disease-specific survival, disease-free survival, and extrahepatic recurrence-free survival (p < 0.05 for all). CONCLUSIONS: LOXL2 expression represents a subgroup of HCCs with more aggressive behavior and is suggested to be a poor prognostic marker in HCC patients.
Sujets)
Humains , Carbonic anhydrases , Carcinome hépatocellulaire , Survie sans rechute , Cellules épithéliales , Matrice extracellulaire , Immunohistochimie , Interleukine-6 , Kératine-19 , Métastase tumorale , Veine porte , PronosticRésumé
ABSTRACT A 89-year-old Black female with a 6-year history of advanced open-angle glaucoma was referred to the Glaucoma Service of the Ophthalmology Department - Federal University of São Paulo (UNIFESP). Best-corrected visual acuity was 20/400 in the right eye and 20/60 in the left eye. Pseudoexfoliation material was observed at the iris border, angle, and the anterior lens surface. Anterior biomicroscopy revealed exfoliation material forming an evident peripheral zone and a central disc separated by a clear intermediate zone on the anterior lens surface OU. Gonioscopy showed an open-angle Sampaolesis's line and whitish material deposits OU. Fundus examination revealed a cup-to-disc ratio of 1.0 OU with peripapillary atrophy. Genetic analysis for single nucleotide polymorphisms of the lysyl oxidase-like 1 gene linked to exfoliation syndrome identified two such single nucleotide polymorphisms, rs1048661 and rs216524.
RESUMO Uma mulher negra de 89 anos com um histórico de seis anos de glaucoma avançado de ângulo aberto avançado foi encaminhada ao Serviço de Glaucoma do Departamento de Oftalmologia da Universidade Federal de São Paulo (UNIFESP). A acuidade visual melhor corrigida era 20/400 no olho direito e 20/60 no olho esquerdo. Material pseudo-exfoliativo foi observado na borda iriana, ângulo e superfície anterior do cristalino. A biomicroscopia de segmento anterior demonstrou material exfoliativo formando uma zona periférica evidente e um disco central separado por uma zona intermediária livre na cápsula anterior do cristalino. A gonioscopia mostrou uma linha de Sampaolesi de ângulo aberto e depósitos esbranquiçados. O exame de fundo de olho revelou disco óptico com escavação total em ambos os olhos com atrofia peripapilar. A análise genética para polimorfismos de nucleotídeo único do gene semelhante à lysyl oxidase-like 1 ligado à síndrome de esfoliação identificou dois desses polimorfismos de nucleotídeo único, rs1048661 e rs216524.
Sujets)
Humains , Femelle , Sujet âgé de 80 ans ou plus , Glaucome capsulaire/génétique , Amino-acid oxidoreductases/génétique , Glaucome capsulaire/imagerie diagnostique , Prédisposition génétique à une maladie , Polymorphisme de nucléotide simple , , Fréquence d'allèleRésumé
Objective To explore the clinical application of the expression of LOXL2 mRNA and Tenascin-C mRNA in tissues for the disease with the bile duct cancer.Methods The serum and clinical data in 35 cases of patients with the bile duct cancer (cancer group) and 28 cases of patients with normal bile duct tissue (control group) were collected,used the real-time fluorescent quantitative PCR (real-time-PCR,RT-PCR) technology to detect the expression of LOXL2 mRNA and TenascinC mRNA in tissues toobserve the relationship between the changes and the bile duct cancer for the two markers.Results The expression of LOXL2 mRNA and Tenascin-C mRNA in tissues in the cancer group were 1.27±0.18 and 1.39±0.19,which of ones in the control group were 0.20±0.06 and 0.23±0.06.In the cancer group,the expression of LOXL2 mRNA and Tenascin-C mRNA in tissues respectively with comparision to those in the control group were significantly higher,the differences had statistical significance(t=52.18,56.87,P＜0.01),which of ones in the cancer group was positively related (r=0.687,P＜0.01).Conclution The expression of LOXL2 mRNA and Tenascin-C mRNA in tissues may be a molecular targets for the disease with the bile duct cancer in the early diagnosis and judgment of progression in the courses of this disease.
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Objective To investigate the expression of LOXL2 protein (lysyl oxidase like-2 protein) and epithelial-mesenchymal transition (EMT) related markers in cholangiocarcinoma tissues and its relation with the malignant features. Methods The expression of LOXL2、E-cadherin and Vimentin protein in 48 cases of cholangiocarcinoma tissues was detected by immunohistochemistry and compared with the clinicopathological data of cholangiocarcinoma. Results The positive expression rate in cholangiocarcinoma was 71% ( 34/48 ) for LOXL2 and 46% ( 22/48 ) for Vimentin, the absent expression rate was 52% (25/48) for E-cadherin. The positive expression rate of LOXL2 was significantly associated with the absent expression of epithelium markers E-cadherin ( r = 0. 394, P < 0. 05 ) and the positive expression of fibroblast markers Vimentin ( r = 0. 406, P < 0. 05 ). There was no correlation between the expression of LOXL2 and patients gender, age, and cancer differentiation, but a significant correlation with tumor metastasis was found ( P < 0. 05 ). Conclusions LOXL2 protein overexpression in cholangiocarcinoma may accelerate invasion of cholangiocarcinoma through induced EMT.