Effect of simulated microgravity on erythroid differentiation of K562 cells and the mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of simulated microgravity on erythroid differentiation of K562 cells and explore the possible mechanism.</p><p><b>METHODS</b>The fourth generation rotating cell culture system was used to generate the simulated microgravity environment. Benzidine staining was used to evaluate the cell inhibition rate, and real-time quantitative PCR (qRT-PCR) was used to detect GATA-1, GATA-2, Ets-1, F-actin, β-Tubulin and vimentin mRNA expressions. The changes of cytoskeleton were observed by fluorescence microscopy, and Western blotting was employed to assay F-actin, β-tubulin and vimentin protein expression levels.</p><p><b>RESULTS</b>Benzidine staining showed that simulated microgravity inhibited erythroid differentiation of K562 cells. K562 cells treated with Hemin presented with increased mRNA expression of GATA-1 and reduced GATA-2 and Ets-1 mRNA expressions. Simulated microgravity treatment of the cells resulted in down-regulated GATA-1, F-actin, β-tubulin and vimentin mRNA expressions and up-regulated mRNA expressions of GATA-2 and Ets-1, and reduced F-actin, β-tubulin and vimentin protein expressions. Exposure to simulated microgravity caused decreased fluorescence intensities of cytoskeletal filament F-actin, β-tubulin and vimentin in the cells.</p><p><b>CONCLUSION</b>Simulated microgravity inhibits erythroid differentiation of K562 cells possibly by causing cytoskeleton damages to result in down-regulation of GATA-1 and up-regulation of GATA-2 and Ets-1 expressions.</p>

Association of two SNPs in 3'UTR of ETS1 gene with systemic lupus erythematosus in a northern Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association between 2 single nucleotide polymorphisms (SNPs) of ETS1 gene and susceptibility to systemic lupus erythematosus (SLE) in a northern Chinese Han population.</p><p><b>METHODS</b>Two SNPs within the ETS1 gene mapped to 11q23 were selected based on HapMap data. Genotyping was conducted with Taqman method in 231 patients with SLE and 474 healthy controls from Qilu Hospital, Shandong and analyzed with PLINK1.07 software. Haplotypes were analyzed with SHEsis software.</p><p><b>RESULTS</b>A statistically significant difference was detected in the distribution of rs1128334 and rs4937333 genotypes between the two groups (all P< 0.01). For rs1128334, the frequency of the minor allele was 0.291 and 0.428 in controls and cases, respectively. For rs4937333, the minor allele frequency was 0.381 and 0.476 in controls and cases respectively. An A-C haplotype was found to be strongly associated with increased risk for SLE, while another haplotype G-C may reduce this risk.</p><p><b>CONCLUSION</b>Our study has suggested that rs1128334 and rs4937333 are strongly associated with the risk for SLE in northern Chinese Han population.</p>

Expression of matrix metalloproteinase-1, E26 transformation-specific-1 in laryngeal carcinoma tissue and the clinical significance / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression of matrix metalloproteinase-1 (MMP-1), E26 transformation-specific-1 (Ets-1) in laryngeal carcinoma and to discuss their relevance and the roles in carcinogenesis and development of laryngeal carcinoma.@*METHOD@#Immunohistochemical technique was used to detect the expression of MMP-1 and Ets-1 protein in 34 tissues of laryngeal carcinoma and 34 para-carcinoma tissues, 15 cases of vocal cord polyps, and the use of the pathological image analysis software, we analysis the results of immunohistochemical semi-quantitatively.@*RESULT@#(1) The expression of MMP-1 and Ets-1 protein in laryngeal carcinoma tissues is obviously higher than that in para-carcinoma and in vocal cord polyps respectively (P 0.05). (2) The expression of MMP-1, Ets-1 protein isn't related to patients' age and sex, tumor size,clinical classification (P > 0.05), but related to pathological grade, clinical stage and lymph nodes metastasis (P < 0.05). (3) There is a positive correlation between the expression of MMP-1 and Ets-1 in laryngeal carcinoma.@*CONCLUSION@#Overexpression of MMP-1 and Ets-1 was observed in laryngeal carcinoma. The high expression of MMP-1 and Ets-1 may contribute to the carcinogenesis and development of laryngeal carcinoma,which is a important value to judge the malignant degree and progress of laryngeal carcinoma.

Expression of natural killer cell development-associated transcription factors in nasal NK/T-cell lymphomas occurring in Chinese population / 中华病理学杂志

<p><b>OBJECTIVE</b>To evaluate the expression and prognostic significance of T-bet and its cofactors EOMES, ETS-1 and MEF [which are transcription factors and responsible for development of natural killer (NK) cells] in the extranodal NK/T-cell lymphoma, nasal type (EN-NK/T-NT).</p><p><b>METHODS</b>The expression status of T-bet, EOMES, ETS-1 and MEF in 40 cases of EN-NK/T-NT occurring in Chinese population was studied by immunohistochemistry and in-situ hybridization (ISH). The clinical relevance was also evaluated. The control cases included 18 cases of peripheral T-cell lymphoma, 10 cases of B-cell lymphoma, 5 cases of normal spleen, 5 cases of normal thymus and 10 cases of nasal mucosal tissues affected by chronic inflammation.</p><p><b>RESULTS</b>The expression levels of T-bet mRNA and protein were high in EN-NK/T-NT (82.5% and 100%, respectively) and in peripheral T cell lymphoma (17/18 and 72.2%, respectively). There was no expression in B-cell lymphoma. The expression of EOMES (80.0% by ISH), ETS-1 (82.5% by ISH) and MEF (62.5% by ISH) was high in EN-NK/T-NT, but not in the control group. The frequency of co-expression of T-bet and EOMES (75%, 30/40) was significantly higher than that of the other genes. Follow-up study showed that the mean and median survival of the 19 cases of EN-NK/ T-NT was 33 months and 10 months, respectively. The five-year survival rate was 10.5%. Statistical analysis showed that only treatment modalities significantly affected the patients' overall survival; and none of the four transcription factors had significant impact on survival. The expression rates of T-bet, EOMES, ETS-1 or MEF had no significant difference between the 9 alive and the 10 dead cases.</p><p><b>CONCLUSIONS</b>The expression of T-bet correlates with the lymphoma types. It is mainly expressed in peripheral NK and T-cell lymphomas. The important functional gene engaged in NK cells development is highly expressed in EN-NK/T-NT. They may play a crucial role in pathogenesis and aggressive biologic behavior.</p>

Expression of bone-related genes in bone marrow MSCs after cyclic mechanical strain: implications for distraction osteogenesis / 国际口腔科学杂志·英文版

<p><b>AIM</b>Understanding the response of mesenchymal stem cells (MSCs) to mechanical strain and their consequent gene expression patterns will broaden our knowledge of the mechanobiology of distraction osteogenesis.</p><p><b>METHODOLOGY</b>In this study, a single period of cyclic mechanical stretch (0.5 Hz, 2,000 microepsilon) was performed on rat bone marrow MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity was examined. The mRNA expression of six bone-related genes (Ets-1, bFGF, IGF-II, TGF-beta, Cbfa1 and ALP) was detected using real-time quantitative RT-PCR.</p><p><b>RESULTS</b>The results showed that mechanical strain can promote MSCs proliferation, increase ALP activity, and up-regulate the expression of these genes. A significant increase in Ets-1 expression was detected immediately after mechanical stimulation, but Cbfa1 expression became elevated later. The temporal expression pattern of ALP coincided perfectly with Cbfa1.</p><p><b>CONCLUSION</b>The results of this study suggest that mechanical strain may act as a stimulator to induce differentiation of MSCs into osteoblasts, and that these bone-related genes may play different roles in the response of MSCs to mechanical stimulation.</p>

Relationship between Ets-1 expression and angiogenesis, clinicopathological features and survival of patients with gastric carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of Ets-1 in gastric carcinoma, para-cancerous tissue and metastatic lymph nodes, and to determine the relationship between Ets-1 expression and clinicopathological features, angiogenesis and survival of patients with gastric carcinoma.</p><p><b>METHODS</b>Gastric carcinoma tissue microarray was used to determine Ets-1 protein expression by SP immunohistochemical staining in 189 advanced gastric cancer, 54 papacancerous tissues, 41 metastatic lymph nodes and 32 control tissues.</p><p><b>RESULTS</b>The positive rates for Ets-1 expression of the carcinoma, paracancerous and control tissues were 71.4%, 29.6% and 18.8%, respectively, with a significant difference among the three groups (P < 0.01). In the cancer tissues, the positive rate of Ets-1 protein expression was significantly associated with depth of invasion and lymph node metastasis (P < 0.01), but not associated with degree of differentiation, Lauren's histological type, sex, age, and size of tumor (P > 0.05). The positive rates for Ets-1 expression of the 41 gastric cancer and 41 metastatic lymph nodes were significantly different (P < 0.05). In metastatic lymph nodes, the positive rate for Ets-1 expression was higher. The MVD in Ets-1 positive tumors was higher than that in the Ets-1 negative tumors, with a significant difference (P < 0.05). Kaplan-Meier survival analysis showed that the survival time of Ets-1-negative patients was longer than that of Ets-1-positive patients (P < 0.05). Cox regression analysis showed that Ets-1 expression was not an independent prognostic factor of gastric carcinoma.</p><p><b>CONCLUSION</b>A higher expression of Ets-1 is involved in carcinogenesis, development, invasion, and metastasis of gastric cancer. Ets-1 plays an important role in angiogenesis in gastric cancer. Ets-1 is a useful marker for predicting the outcome for patients with gastric carcinoma, though it is not an independent prognostic indicator.</p>

Relationship between the expressions of KaI1, nm23, ETS-1, VEGF and microvascular density and clinical significance in nasopharyngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the expressions of KAI1, nm23, ETS-1, vascular endothelial growth factor (VEGF) and microvascular density (MVD) and lymph node metastasis and prognosis in nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>The Envision immunohistochemical method was used to detect the expressions of KAI1, nm23, ETS-1 and VEGF in 50 cases of non-keratinizing carcinoma (NKC) with cervical lymph node metastasis, 30 cases of NKC without cervical lymph node metastasis at the primary diagnoses and 30 cases of non-tumor nasopharyngeal tissues (NP). The microvascular density was counted by immunostaining with CD34.</p><p><b>RESULTS</b>(1) The expression rates of KAI1 and nm23 protein in NKC with cervical lymph node metastasis group and without cervical lymph node metastasis group and NP group increased successively , the difference being significant (P < 0.05); The expression rates of ETS-1 and VEGF protein in NKC with cervical lymph node metastasis group and without cervical lymph node metastasis group and NP group increased successively, the difference being significant (P < 0.05). (2) In 80 NKC cases, the MVD was respectively lower in KAI1 and nm23 protein positive groups than those in the negative groups (P < 0.05); the MVD was respectively higher in ETS-1 and VEGF protein positive groups than those in the negative groups (P < 0.05 ). (3) There was significant difference between the MVD, the number of NKC without cervical lymph node metastasis cases in the single expression of KAI1 or nm23 protein and in common expression of KAI1 and nm23 protein (P < 0.05), in the same as between the single expression of ETS-1 or VEGF protein and in common expressions of ETS-1 and VEGF protein (P < 0.05). (4) There was positive correlation between the expressions of KAI1 and nm23 protein (P < 0.01), as well as between the expressions of ETS-1 and VEGF protein (P < 0.01). (5) the 5-year survival rates of the patients correlated with cervical lymph node metastasis and the expressions of KAI1, nm23, ETS-1 and VEGF proteins in NKC (P < 0.05).</p><p><b>CONCLUSIONS</b>The expressions of KAI1, nm23, ETS-1 and VEGF proteins were highly related to MVD in NPC,cervical lymph node metastasis and prognosis. They might be considered to be reference indicator for evaluating the cervical lymph node metastasis and prognosis of NPC.</p>

Cross-talk between c-Jun/Ets1 involved in EB virus-encoded latent membrane protein 1 regulates expression of matrix metalloproteinase-9 in nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate effect of AP-1 and Ets binding site adjacent to matrix metalloproteinase-9 (MMP-9) promoter on activation of MMP-9 transcription of nasopharyngeal carcinoma cells transfected with EBV-encoded latent membrane protein 1 (LMP1), and to ascertain if cross-talk between c-Jun and Ets1 is involved in LMP1-regulating expression of MMP-9.</p><p><b>METHODS</b>Site-directed mutagenesis technique was used to establish a series of mutants, including MMP-9-CAT-Ets(-540)mt, MMP-9-CAT-AP-1(-533)mt and MMP-9-CAT-AP-1(-533)/Ets(-540)mt. After the mutants were transfected into LMP1-expressing NPC HNE2 cells regulated by Tet-on system (pTet-on-LMP1 HNE2), CAT activity of these mutants were assayed with induction of LMP1. With blockade of c-Jun or Ets1 antisense oligonucleotides, the activity of MMP-9 induced by LMP1 was assayed with gelatin zymography.</p><p><b>RESULTS</b>The CAT activity of MMP-9-Ets(-540)mt-CAT, MMP-9-AP-1(-533)mt-CAT, MMP-9-AP-1(-533)/Ets(-540) mt-CAT decreased significantly compared to MMP-9-CAT wt. After blockade with c-Jun or Ets1 antisense oligonucleotides, activity of MMP-9 induced by LMP1 decreased significantly, especially with combined blockade of c-Jun and Ets1.</p><p><b>CONCLUSION</b>The results suggest that transcription factor AP-1 and Ets play an crucial role in activation of MMP-9 transcription induced by LMP1, and cross-talk between c-Jun/Ets1 is involved in expression of MMP-9 mediated by LMP1.</p>

In vitro study of the effects of estrogen receptor beta expression on the biological behavior of a human breast cancer cell line / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study effects of estrogen receptor beta (ER beta) on the biological behavior of a human breast cancer cell line MDA-MB-435.</p><p><b>METHODS</b>Human ER beta cDNA was introduced into MDA-MB-435 cells by stable transfection. Effects of ER beta expression on cell proliferation and invasion were investigated by MTT, flow cytometry and transwell techniques. Cyclin A, cyclin E, cyclin D1, p21, MMPs, Ets-1, VEGF and b-FGF were detected by RT-PCR and/or Western blot or gelatin zymography.</p><p><b>RESULTS</b>ER beta was shown to be able to significantly increase the proliferation and invasion of MDA-MB-435 cells in an estradiol-independent manner. The S phase distribution of the cells with ER beta overexpression was 46.8%, significantly higher than that of wild type (29.9%) and mock transfected cells (27.6%) (P = 0.01). In ER beta transfected cells, the expression of p21 decreased by 33.3% at mRNA level (P = 0.03) and by 47.4% at protein level (P = 0.02), respectively. The expression of MMP-9 increased by 91.3% at mRNA level (P < 0.01) and its activity was up-regulated by 67.3% (P = 0.02). Furthermore, the mRNA and protein levels of Ets-1 increased 62.2% (P = 0.01) and 51.0% (P = 0.01), respectively. No significant difference was observed in the mRNA levels of cyclin A, cyclin E, cyclin D1, MMP-1, MMP-2, MMP-7, VEGF and b-FGF among these cells.</p><p><b>CONCLUSION</b>ER beta can enhance proliferation and invasion of breast cancer cells. Down-regulation of p21 and up-regulation of MMP-9 and Ets-1 may be involved in its mechanisms.</p>

EB virus encoded latent membrane protein 1 mediated expression of transcription factor Ets-1 in nasopharyngeal carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To elucidate the expression of tanscription factor Ets-1 mediated by EB virus encoded latent membrane protein 1 (LMP1) in nasopharyngeal carcinoma (NPC) cells.</p><p><b>METHODS</b>LMP1-expressing NPC HNE2 cells regulated by Tet-on system (pTet-on-LMP1 HNE2) were used. Expression of LMP1 and Ets-1 was observed after induction with Doxycycline (Dox). Expression of Ets-1 mRNA and protein was detected by RT-PCR and Western blot, respectively. The phosphorylation level of Ets-1 protein was examined by co-immunoprecipitation. The DNA binding activity of Ets-1 was detected by electrophoretic-mobility shift assay (EMSA).</p><p><b>RESULTS</b>After induction with Dox in pTet-on-LMP1 HNE2 cells, to some extent, the expression of Ets-1 mRNA and protein, its phosphorylation level and DNA binding activity were increased with enhancement of LMP1 expression.</p><p><b>CONCLUSION</b>LMP1 induces transcriptional activation and expression of Ets-1 which may contribute to the development of NPC.</p>

Expression and significance about VEGF, KDR, MMP-1, and transcription factor Ets-1 in human cervical carcinoma / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the expression of VEGF, KDR, MMP-1, and its transcription factor Ets-1 on the interstitial neovasculogenesis in human cervical carcinoma on molecular level, which may provide further theoretic bases on judgement of prognosis and explain interregularity between neovasculagenetic factors.</p><p><b>METHODS</b>VEGF, KDR, MMP-1, and Ets-1 were detected in 87 cervical carcinomas by in situ hybridization and immunohistochemistry. The survival curves for patients with detected tumors were compared with the curves for those without tumors. Multivariate analyses were performed with Pearson analysis.</p><p><b>RESULTS</b>VEGF mRNA and its protein were mainly expressed in cytoplasms of tumor cells, positive rate was 78.6% (68/87) and 70.4% (61/87) respectively. KDR mRNA and its protein were mainly expressed in vascular endothelial cells, as correlation coefficient between KDR and VEGF was 0.892. Over expression of MMP-1 was seen in both endothelial cells and tumor cells, especially in hyperplasia of endothelial cells. Ets-1 was mainly expressed in both endothelial cells and tumor cells, correlation coefficient between Ets-1 and KDR, MMP-1, was 0.900 and 0.873, respectively. Four factors of above were highly related with tumor differentiation, lymph nodes metastasis as well as 5 years survival rate.</p><p><b>CONCLUSIONS</b>VEGF, KDR, MMP-1, and Ets-1 are important factors on neovasculogenesis in cervical carcinoma, which may be considered to be reference indicator for determining biology behavior of cervical carcinoma, and may provide further theoretic bases on antineovasculagenetic therapy.</p>

Ets oncogene family.

First member of Ets gene family was discovered a decade ago by studying avian erythroblastosis virus, E26. This virus encodes a tripartite protein gag-myb-ets with a molecular weight of 135 kDa. Subsequently, a series of cellular Ets genes were isolated (Ets-1, Ets-2, Erg, Elk-1, Sap-1, PEA-3, PU.1, Fli-1, Pok/Yan, Etv-1 etc.). These genes share sequence homology to E26 Ets gene (v-ets or viral ets). Ets genes are highly conserved in phylogenetically divergent species from Drosophila to man. Mammalian Ets genes are located on different chromosomes. Ets gene products are transcriptionally active sequence-specific DNA binding proteins and are differentially regulated. Ets genes are involved in certain chromosomal translocations leading to the formation of chimeric fusion proteins that are associated with certain leukemias and soft tissue cancers. Ets genes also have a role in T-cell development and molecular and genetic analysis of Down Syndrome patients have implicated the human Ets-2 and Erg genes in the disease. Down Syndrome afflicted patients have immunologic and thymic disorders as well as a greater risk for leukemic disease. Thus, Ets genes having homology to viral oncogenes, may be instrumental in regulating cellular growth and differentiation, as well as organismal development. Alteration of these genes and their products may cause deregulation of normal cell growth and differentiation and result in a disease.