Effect of the regulator of G-protein signaling 2 on the proliferation and invasion of oral squamous cell carcinoma cells and its molecular mechanism / 华西口腔医学杂志

OBJECTIVES@#This study aims to investigate the effect of the regulator of G-protein signaling 2 (RGS2) on the proliferation and invasion of oral squamous cell carcinoma (OSCC) cells and its potential molecular mechanism. Metho⁃ds The expression status and clinical significance of RGS2 in head and neck squamous cell carcinomas and matched adjacent normal tissues were evaluated using TCGA database. Three OSCC cell lines (i.e., SCC-9, Cal27, and Fadu) were overexpressed with RGS2, and the effect of RGS2 on cell proliferation and invasion was determined using the Transwell, clone formation, and cell counting kit (CCK)-8 assays. Moreover, the yeast two-hybrid scree-ning and co-immunoprecipitation (Co-IP) assays were conducted to detect the correlation of RGS2, four and a half LIM domains protein 1 (FHL1), and damage DNA-binding protein 1 (DDB1).@*RESULTS@#The expression level of RGS2 in OSCC was significantly lower than that in matched adjacent normal tissues (@*CONCLUSIONS@#RGS2 plays an important role in the inhibition of OSCC proliferation and invasion. The structure stability of RGS2 is competitively regulated by FHL1 and DDB1.

LIM domain only 1: an oncogenic transcription cofactor contributing to the tumorigenesis of multiple cancer types / 中华医学杂志(英文版)

The LIM domain only 1 (LMO1) gene belongs to the LMO family of genes that encodes a group of transcriptional cofactors. This group of transcriptional cofactors regulates gene transcription by acting as a key "connector" or "scaffold" in transcription complexes. All LMOs, including LMO1, are important players in the process of tumorigenesis. Unique biological features of LMO1 distinct from other LMO members, such as its tissue-specific expression patterns, interacting proteins, and transcriptional targets, have been increasingly recognized. Studies indicated that LMO1 plays a critical oncogenic role in various types of cancers, including T-cell acute lymphoblastic leukemia, neuroblastoma, gastric cancer, lung cancer, and prostate cancer. The molecular mechanisms underlying such functions of LMO1 have also been investigated, but they are currently far from being fully elucidated. Here, we focus on reviewing the current findings on the role of LMO1 in tumorigenesis, the mechanisms of its oncogenic action, and the mechanisms that drive its aberrant activation in cancers. We also briefly review its roles in the development process and non-cancer diseases. Finally, we discuss the remaining questions and future investigations required for promoting the translation of laboratory findings to clinical applications, including cancer diagnosis and treatment.

La leptina promueve la expresión de Hic-5 y la formación de puntos de actina por la vía dependiente de FAK-Src en células epiteliales mamarias MCF10A / Leptin induced Hic-5 expression and actin puncta formation by the FAK/Src-dependent pathway in MCF10A mammary epithelial cells

Resumen Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial- Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.

Abstract Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.

Relationship between Testin expression and clinicopathological characteristics in nasopharyngeal carcinoma patients / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression of Testin gene, and analyze its possible relationship with the clinicopathological features of human nasopharyngeal carcinoma.@*METHOD@#The expression of Testin in nasopharyngeal carcinoma tissues were detected by immunohistochemistry methods, semi-quantitative reverse transcription polymerase chain reaction and Western blot. The correlations of Testin to clinicopathologic features of nasopharyngeal carcinoma were analyzed.@*RESULT@#The positive expression rate of Testin in NPC biopsy tissue was 37.8% (17/ 45), while it was 88.9% (40/45) in the normal tissue; The expression of Testin mRNA was significantly lower than that in the normal tissue (P 0.05); but it had significant correlation with lympho node metastasis, distant metastasis and differentiation degree.@*CONCLUSION@#The decreased expression of Testin gene may play an importmant role in the development of nasopharyngeal carcinoma. And thus Testin gene might be a novel candidate of tumor-suppressor. It may be an objective marker for prognostic factor and malignant level for nasopharyngeal carcinoma.

Inhibitory effect of KyoT2 overexpression on proliferation and migration of airway smooth muscle cells in mice with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of KyoT2 on the proliferation and migration of airway smooth muscle cells (ASMCs) in mice with asthma.</p><p><b>METHODS</b>Ovalbumin (OVA) was used to establish the asthmatic model of airway remodeling in BALB/c mice. ASMCs were isolated and cultured, and primarily cultured ASMCs were used as the control group. The expression of KyoT2 in ASMCs was measured in the control and asthma groups. After the ASMCs from asthmatic mice were transfected with pCMV-Myc (empty vector group) or pCMV-Myc-KyoT2 plasmid with overexpressed KyoT2 (KyoT2 expression group) for 48 hours, RT-PCR and Western blot were used to measure the mRNA and protein expression of KyoT2, the MTT assay and BrdU assay were used to measure the proliferation of ASMCs, and Transwell assay was used to measure the migration of ASMCs. Western blot was used to determine the effect of KyoT2 overexpression on the protein expression of RBP-Jκ, PTEN, and AKT.</p><p><b>RESULTS</b>Compared with the control group, the asthma group had significantly downregulated expression of KyoT2 in ASMCs, and the KyoT2 expression group had significantly upregulated expression of KyoT2 in ASMCs (P<0.05). Compared with the empty vector group, overexpressed KyoT2 significantly inhibited cell proliferation and migration, downregulated the expression of RBP-Jκ and AKT, and upregulated the expression of PTEN.</p><p><b>CONCLUSIONS</b>Overexpressed KyoT2 can inhibit the proliferation and migration of ASMCs through the negative regulation of RBP-Jκ/PTEN/AKT signaling pathway.</p>

Expression and significance of HGAL and LMO2 in follicular lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To evaluate the diagnostic value of HGAL and LMO2 expression and compare with CD10 and bcl-6 in follicular lymphoma (FL).</p><p><b>METHODS</b>63 cases of FL were collected from Guangdong General Hospital. The expression of HGAL, LMO2, CD10 and bcl-6 was assessed by immunohistochemistry.</p><p><b>RESULTS</b>The expression rates of HGAL, LMO2, CD10 and bcl-6 were 98.4% (62/63), 82.5% (52/63), 82.5% (52/63) and 87.3% (55/63), respectively. The expression rate of HGAL was higher than those of LMO2, CD10 and bcl-6, but the differences were not significant (P>0.05). There was no significant difference in HGAL, LMO2 and bcl-6 expression among FL1, FL2 and FL3 cases. The CD10 expression rate of FL1-3A cases was significantly higher than that of FL3B cases(P<0.01).</p><p><b>CONCLUSIONS</b>HGAL and LMO2, especially HGAL, can be used in FL particularly high grade FL as useful germinal center marker.</p>

Mutation analysis of large tumor suppressor genes LATS1 and LATS2 supports a tumor suppressor role in human cancer

In recent years, human cancer genome projects provide unprecedented opportunities for the discovery of cancer genes and signaling pathways that contribute to tumor development. While numerous gene mutations can be identified from each cancer genome, what these mutations mean for cancer is a challenging question to address, especially for those from less understood putative new cancer genes. As a powerful approach, in silico bioinformatics analysis could efficiently sort out mutations that are predicted to damage gene function. Such an analysis of human large tumor suppressor genes, LATS1 and LATS2, has been carried out and the results support a role of hLATS1//2 as negative growth regulators and tumor suppressors.

Expression and clinical significance of LIMD-1 gene in adult patients with acute leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>The reduced expression or expression deletion of LIM domain-containing protein 1(LIMD1) gene associates with the occurrence of various solid tumors, while its role in adult acute leukemia (AL) has been rarely reported, this study was to detect LIMD1 expression in adult patients with AL and to evaluate its correlation with the different clinical and laboratorial data.</p><p><b>METHODS</b>The expression levels of LIMD1 were determined by real-time quantitative PCR (RT-qPCR), LIMD1 transcripts were measured by using a relative quantification with GAPDH as a housekeeping gene, and the relationship between its expression levels and clinical parameters (sex, age, subtype, leukocyte count, leukemic blasts) was investigated by statistical analysis.</p><p><b>RESULTS</b>The LIMD1 expression was reduced in de novo AL patients as compared with normal controls and complete remission patients(P < 0.01). In univariate analysis, LIMD1 associated with age and leukocyte count (P = 0.011, 0.035 respectively). LIMD1 decreased along with increasing age and leukocyte count in de novo AL patients, the LIMD1 expression levels in de novo AL patients with age ≥ 60 years old were lower than that in group of patients <60 years, and which were significantly lower in the leukocyte count ≥ 100×10(9)/L compared to leukocyte count < 100×10(9)/L. there was no statistically significant association between LIMD1 expression and sex, subtype, and leukemic blasts of patients.</p><p><b>CONCLUSIONS</b>The LIMD1 gene may be involved in the pathogenesis and progression of adult AL, and may be used as an indicator of prognosis evaluation.</p>

Ribotrap analysis of proteins associated with FHL3 3'untranslated region in glioma cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To screen the proteins associated with four-and-a-half LIM domains 3 (FHL3) 3' untranslated region (3'UTR) in glioma cells.</p><p><b>METHODS</b>Western blot was adopted to detect the regulatory effect of poly(C)-binding protein 2 (PCBP2) on FHL3. Biotin pull-down and sliver staining were employed to screen and verify the candidate binding proteins of FHL3 3'UTR. Then liquid chromatography-tandem mass spectrometry (LC-MS/MS) and molecule annotation system were used to identify and analyze the candidate binding proteins. Immuno- precipitation was conducted to study the interaction between PCBP2 and polypyrimidine tract-binding protein 1 (PTBP1), a binding protein identified by LC-MS/MS.</p><p><b>RESULTS</b>PCBP2 could bind to FHL3 mRNA 3'UTR-A and inhibited the expression of FHL3 in T98G glioms cells. 22 candidate binding proteins were identified. Among them, there were 11 RNA binding proteins, including PCBP2. PTBP1 associated with FHL3 mRNA 3'UTR and interacted with PCBP2 protein.</p><p><b>CONCLUSIONS</b>PCBP2 and PTBP1 can both associate with FHL3 mRNA 3'UTR through forming a protein complex.</p>

Prognostic value of clinical characteristics and immunophenotypic biomarkers in 115 patients with primary central nervous system lymphoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Clinical outcome in patients with primary central nervous lymphoma (PCNSL) is variable and poorly predictable. This study investigated the association of clinical features and immune markers with prognosis of patients with PCNSL.</p><p><b>METHODS</b>One hundred and fifteen newly diagnosed PCNSL patients at the study institution were considered eligible for this study. Clinical characteristics and biochemical assay data were collected. Immunohistochemical staining of Cyclin D3, Cyclin E, Foxp1, and LMO2 were performed. All cases were followed-up regularly.</p><p><b>RESULTS</b>The common sites of involvement were frontal lobe (54.8%) and thalamus (16.5%). Diffuse large B-cell lymphoma composed of 96.5% of the cases. The median overall survival was 22 (4 - 41) months, and the 5-year survival rate was 22.8%. Age > 65 years, serum globulin > 40 g/L, large size of tumor, lymphocyte count ≥ 1 × 10(9)/L, and expression of Cyclin D3 and Cyclin E were associated with poor prognosis of PCNSL. Expressions of Foxp1, LMO2, and CD44 were not related to the survival. Expression of Cyclin E, large tumor size, and high serum globulin were independent prognostic factors for PCNSL.</p><p><b>CONCLUSIONS</b>PCNSL prognosis is relatively poor. Age, high tumor burden, higher lymphocyte count, expression of Cyclin D3, and Cyclin E are inferior prognostic factors for PCNSL.</p>

Recombinant human bone morphogenetic protein-2 down-regulates LASP1 and up-regulates ferritin during osteogenic differentiation of beagle dog bone marrow mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify the proteins involved in osteogenic differentiation of beagle dog bone marrow mesenchymal stem cells (BMSCs) and explore its possible regulation mechanism of osteogenic differentiation of BMSCs.</p><p><b>METHODS</b>Cultured beagle dog BMSCs were induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) for 7 days. The differentially expressed proteins between cells with osteogenic differentiation and control cells were identified by proteomics analysis based on two-dimensional gel electrophoresis. Q-PCR and Western blotting were used to verify the interested protein of LASP1, ferritin light chain and heavy chain.</p><p><b>RESULTS</b>Osteogenic differentiation was induced successfully in the BMSCs. Twenty differentially expressed proteins were identified by proteomic analysis, including 9 down-regulated and 11 up-regulated ones. Q-PCR and Western blotting demonstrated a significant reduction of LASP1 expression and significant up-regulation of ferritin in the BMSCs after a 7-day induction with rhBMP-2.</p><p><b>CONCLUSION</b>LASP1, which plays an important role in the regulation of the activity of the cytoskeleton, and ferritin, an important molecule in cellular iron homeostasis, can be critical in the osteogenic differentiation of beagle dog BMSCs induced by rhBMP-2.</p>

Expression of microRNA-223 in lymphocytic leukemia cells and its action mechanism / 中国实验血液学杂志

This study was aimed to investigate the expression level and mechanism of microRNA-223 and LMO2 in acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) cells and the mechanism. MicroRNA-223 mimics was transfected to increase the expression of MicroRNA-223 in the lymphocytes sorted by ficoll separation from the bone marrow mononuclear cells (BMMNC) of ALL and CLL patients. MicroRNA-223 inhibitor was transfected to decrease the expression of the MicroRNA-223 in the lymphocytes of normal controls. Then the expression of the MicroRNA-223 and LMO2 in transfected lymphocytes before and after cultivating for 72 hours were detected by RT-PCR, the apoptosis and cell cycle of these cells were measured by flow cytometery. The results indicated that before the transfection, the expression of MicroRNA-223 in ALL and CLL cells was (433.11 ± 144.88), which was significantly lower than that in the normal lymphocyte (949.59 ± 267.39); the expression of LMO2 was (807.10 ± 238.41), which was significantly higher than that in the normal lymphocytes (455.32 ± 176.83) (P < 0.05); after the transfection, the expression of MicroRNA-223 was (571.86 ± 142.00) in ALL and CLL cells, which was significantly higher than that before transfection (P < 0.05), but the expression of LMO2 was significantly lower than that before transfection (651.97 ± 230.12) (P < 0.05); in the normal control the expression of MicroRNA-223 obviously decreased (646.32 ± 172.93) (P < 0.05), the expression of LMO2 was significantly increased (541.27 ± 158.86.2) (P < 0.05). After transfection, the cell cycle G1/G2 phase and apoptosis changed in ALL and CLL cells. Before transfection the cell ratio in cell cycle G1/G2 phase was (94.75 ± 3.15)%, the cell ratio in S phase was (5.14 ± 3.12)%; after transfection the cell ratio in cell cycle G1/G2 phase was (97.03 ± 2.08)% and obviously increased (P < 0.05), the cell ratio in S phase was (2.97 ± 2.08)% and significantly decreased (P < 0.05). Before transfection the apoptosis rate was (54.47 ± 8.72)%, and obviously was higher than that after transfection (60.48 ± 8.81)%. And in the normal control, the cell ratio in G1/G2 phase was significantly higher than that after transfection [(96.73 ± 2.26)%, (94.55 ± 2.77)%, P < 0.05)], and the cell ratio in S phase was significantly increased [(3.25 ± 2.26)%, (5.45 ± 2.77)% (P < 0.05)]. The apoptotic rate in the ALL and CLL patients was significantly higher than that after the transfection [(54.47 ± 8.72)% vs (60.48 ± 8.81)%, respectively (P < 0.05)]. The apoptotic rate in the normal control was significantly lower than that after the transfection [(59.02 ± 10.20)%, (51.96 ± 10.20)%, respectively (P < 0.05)]. It is concluded that the expression of MicroRNA-223 decreases, and the expression of LMO2 increases in lymphocytic leukemia cells which leads to the lymphocytes over-proliferation and abnormal apoptosis, thus may be one of pathogenesis in lymphocytic leukemia.

Effect of LMO2 overexpression in hemangioblast on its proliferation and hematopoietic differentiation / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate function of the Lim-only protein(LMO2) in hemangioblast generated from murine embryonic stem cells differentiation to hematopoietic cells.</p><p><b>METHODS</b>The hemangioblast-specific expression vector with lmo2 or green fluorescence protein gene was constructed, respectively. The murine embryonic stem cells were transfected by the hemangioblast-specific expression vectors. The neomycin-resistance ES cell clones were obtained after having been screened by G418. The cell clones were spontaneously differentiated into embryo bodies(EB) containing hemangioblast.Expression of the hematopoietic genes was investigated by real-time reverse transcription-ploymerase chain reaction during EB differentiation.For the EB cells, blast-cloning forming cells analysis and blood-colony forming unit analysis were then performed, respectively. The numbers of the blasts were counted during hematopoietic differentiation.</p><p><b>RESULTS</b>The hemangioblast-specific expression vector with lmo2 or green fluorescence protein was transfected into ES cells.The neomycin-resistance ES cells generated EBs from 2.5 days to 10 days.Real time reverse transcription-ploymerase chain reaction analysis indicated that overexpression of lmo2 increased the expression of hematopoietic genes(gata1, tal1, Β-h1, and Β-major globin) during EB formation.Blast-cloning forming cells analysis showed that the numbers of the blasts generated by ES/lmo2 was 2-or 3-fold than those in the controls.The total numbers of the blood-colony forming unit or the numbers of the erythrocyte colony-forming unit generated by ES/lmo2 were 2.5 times or 3 times, respectively, when compared with the controls.</p><p><b>CONCLUSION</b>LMO2 enhances the proliferation and differentiation of hemangioblasts.</p>

Expression of LIM mineralization protein-1 in the apical papilla and dental pulp tissues of human teeth / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To examine the expression of LIM mineralization protein-1 (LMP-1) in the apical papilla and dental pulp tissues of human immature permanent teeth and to investigate the role of LMP-1 in the development and maturation of pulp-dentin complex.</p><p><b>METHODS</b>Forty-eight healthy premolars in need of extraction for orthodontic treatment were obtained with 24 immature permanent teeth and 24 mature permanent teeth. After extraction, the apical papilla was detached from the dental pulp in the immature permanent tooth and the dental pulp of mature permanent tooth was rapidly removed. The samples were divided into 3 groups: group 1, apical papilla of immature permanent teeth (root formed 2/3 of its full length); group 2, dental pulp tissues of immature permanent teeth; group 3, dental pulp tissues of mature permanent teeth. There were 24 samples for each group.Half of them were used for reverse transcriptien-PCR (RT-PCR) detection, and the other half were used for Western blotting detection. Band intensities were quantified using Meta Morph software 6.2.6 and subsequently normalized by dividing the band gray value of the target gene by the intensity of its corresponding β-actin. Two-sample t test was used to analyze the difference of expression intensity between group 1 and group 2 as well as group 2 and group 3 with SPSS 13.0 software package.Statistical significance was established as P < 0.05.</p><p><b>RESULTS</b>As indicated by RT-PCR, LMP-1 expressed in the apical papilla, dental pulp of immature permanent teeth and the dental pulp of mature permanent teeth were 0.25 ± 0.09, 0.46 ± 0.24 and 0.31 ± 0.10 respectively. The expression intensity of LMP-1 mRNA in the dental pulp tissues of human immature permanent teeth were significantly higher than that in the apical papilla tissues(t = 2.92) and that in the dental pulp tissues of human mature permanent teeth (t = 2.31) (P < 0.05). As indicated by Western blotting, LMP-1 expressed in the three groups were 0.33 ± 0.08, 0.82 ± 0.10 and 0.52 ± 0.19 respectively. The expression intensity of LMP-1 protein in the dental pulp tissues of human immature permanent teeth were significantly higher than that in the apical papilla tissues(t = 3.33) and that in the dental pulp tissues of human mature permanent teeth (t = 3.11) (P < 0.05).</p><p><b>CONCLUSIONS</b>LMP-1 were positively expressed in all the samples including apical papilla of immature permanent teeth, dental pulp of immature and mature permanent teeth at the level of both mRNA and protein, but with different intensity. LMP-1 could play an important role in the development and maturation of pulp-dentin complex.</p>

Effects of recombinant gene lentivirus containing LIM mineralization protein-1 on proliferation effect and expression of bone marrow mesenchymal stem cells in rats / 中国骨伤

<p><b>OBJECTIVE</b>To explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.</p><p><b>METHODS</b>Six clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.</p><p><b>RESULTS</b>1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).</p><p><b>CONCLUSION</b>There is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.</p>

Reduced expression of PDLIM4 gene correlates with good prognosis in acute myeloid leukemia / 中国实验血液学杂志

This study was aimed to investigate the expression pattern of gene PDLIM4 (PDZ and LIM domain 4) and analyze its clinical correlation with the patients suffered from acute myeloid leukemia (AML). The expression pattern of PDLIM4 in AML was detected by using EvaGreen real-time quantitative PCR (RQ-PCR). The results showed that the PDLIM4 transcript significantly decreased in 94 AML patients, compared with 21 controls (P < 0.01). The decrease of PDLIM4 transcript was found in 42 (45%) AML patients. PDLIM4 low-expression occurred among the subtypes of M1/M2/M3 more frequently than that in M4/M5/M6 (56% vs 20%, P < 0.01). AML patients with PDLIM4 low-expression had an overall survival (OS) higher than that in AML patients without PDLIM4 low-expression (P < 0.05). Analysis with receiver operating characteristic curve (ROC) displayed that PDLIM4 expression possesses the diagnostic value to differentiate AML from controls, with ROC curve area of 0.865 (95% CI: 0.801-0.930). It is concluded that reduced PDLIM4 expression is a common and favorable event for the good prognosis in AML, and can be used as a potential diagnostic biomarker of cancer.

Effect of Pinch-3 gene interference of glomerular podocytes on cell morphology and cell traction force / 生物医学工程学杂志

Pinch-3 protein is an important constituent of cell membranes, which directly affects the cell morphology and mechanical properties. We observed and compared the change of morphology and cell traction force of glomerular podocytes before and after Pinch-3 gene inhibition by gene interference technology in this experiment. We found that a number of pores appeared on the cell surface, and the cell projected area were increased at the same time, with an approximate average about an increase of 40% after Pinch-3 gene inhibition. The results showed that the cell traction force of glomerular podocytes was significantly reduced, with an approximate average decrease of 40%, the maximum value of the cell traction force was reduced and the distribution of cell traction force became dispersive. All this suggested that after Pinch-3 gene inhibition, some pores created on the cell surface influenced the physical properties of glomerular podocytes and then affected the cell projected area and influenced the formation and distribution of cell traction force of the glomerular podocytes as well.

Regulation of E-cadherin promoter activity by LMO2 impact on the progression and metastasis of prostate cancer / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the abnormal expression of the proto-oncogene LMO2 affect the progression and metastasis mechanism of prostate cancer.</p><p><b>METHODS</b>A series of reporter gene expression vectors carrying different lengths and point mutations of E-cadherin promoter were constructed. These plasmids were separately co-transfected with LMO2 into Lncap cells and the luciferase activity was detected after 24 h.</p><p><b>RESULTS</b>The overexpression of LMO2 could significantly inhibit the activity of luciferase reporter gene of E-cadherin promoter about 50%. Truncated and point mutation study showed that this was mainly through E-box sites in the promoter region -204/-198.</p><p><b>CONCLUSION</b>The proto-oncogene LMO2 can affect the progression and metastasis mechanism of prostate cancer by transcriptional inhibition of E-cadherin.</p>

Hypermethylation of testis derived transcript gene promoter significantly correlates with worse outcomes in glioblastoma patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Glioblastoma is the most common and lethal cancer of the central nervous system. Global genomic hypomethylation and some CpG island hypermethylation are common hallmarks of these malignancies, but the effects of these methylation abnormalities on glioblastomas are still largely unclear. Methylation of the O6-methylguanine-DNA methyltransferase promoter is currently an only confirmed molecular predictor of better outcome in temozolomide treatment. To better understand the relationship between CpG island methylation status and patient outcome, this study launched DNA methylation profiles for thirty-three primary glioblastomas (pGBMs) and nine secondary glioblastomas (sGBMs) with the expectation to identify valuable prognostic and therapeutic targets.</p><p><b>METHODS</b>We evaluated the methylation status of testis derived transcript (TES) gene promoter by microarray analysis of glioblastomas and the prognostic value for TES methylation in the clinical outcome of pGBM patients. Significance analysis of microarrays was used for genes significantly differently methylated between 33 pGBM and nine sGBM. Survival curves were calculated according to the Kaplan-Meier method, and differences between curves were assessed using the log-rank test. Then, we treated glioblastoma cell lines (U87 and U251) with 5-aza-2-deoxycytidines (5-aza-dC) and detected cell biological behaviors.</p><p><b>RESULTS</b>Microarray data analysis identified TES promoter was hypermethylated in pGBMs compared with sGBMs (P < 0.05). Survival curves from the Kaplan-Meier method analysis revealed that the patients with TES hypermethylation had a short overall survival (P < 0.05). This abnormality is also confirmed in glioblastoma cell lines (U87 and U251). Treating these cells with 5-aza-dC released TES protein expression resulted in significant inhibition of cell growth (P = 0.013).</p><p><b>CONCLUSIONS</b>Hypermethylation of TES gene promoter highly correlated with worse outcome in pGBM patients. TES might represent a valuable prognostic marker for glioblastoma.</p>

Association of prostate cancer with PDLIM5, SLC22A3 and NKX3-1 in Chinese men / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the association of prostate cancer (PCa) with PDLIM5 (rs17021918, T), SLC22A3 (rs9364554, C) and NKX3-1 (rs1512268, A) in Chinese men.</p><p><b>METHODS</b>We included 124 PCa patients and 138 normal controls in this study, compared the alleles and genotypes of PDLIM5 (rs17021918, T) , SLC22A3 (rs9364554, C) and NKX3-1 (rs1512268, A) of the two groups, and explored the association of each of the genes with the age, body mass index (BMI), Gleason score, PSA level and tumor stage of the patients. We analyzed the gene-gene interaction using the multifactor dimensionality reduction method (MDR).</p><p><b>RESULTS</b>There were no statistically significant differences in the frequency distribution of the risk alleles and genotypes of PDLIM5, SLC22A3 and NKX3-1 between the case and control groups (P > 0.05), nor were the three gene loci significantly associated with the age, Gleason score, PSA level and pathological grade of the PCa patients (CP < 0.05). MDR analysis showed no interaction between PDLIM5 and NKX3-1, but tree-diagram analysis revealed a possible synergistic action of the two polymorphism loci.</p><p><b>CONCLUSION</b>PCa might not be associated with PDLIM5 (rs17021918,T), SLC22A3 (rs9364554,C) and NKX3-1 (rs1512268,A) in Chinese men. However, PDLIM5 and NKX3-1 might have a synergistic action on the risk PCa.</p>