MiR-195 inhibits migration, invasion and epithelial-mesenchymal transition (EMT) of endometrial carcinoma cells by targeting SOX4

MicroRNAs (miRNAs) have been identified as potential biomarkers for endometrial carcinoma (EC) diagnosis, prognosisand therapy. The purpose of the present study was to investigate the detailed role and molecular mechanism of miR-195 inEC metastasis. qRT-PCR assay was performed to assess the expression of miR-195 and SRY-related high-mobility groupbox 4 (SOX4) mRNA in EC tissues and cells. The levels of N-cadherin, Vimentin, E-cadherin and SOX4 protein weredetermined by western blot. SOX4 protein expression in EC tissues was also determined by Immunohistochemistry (IHC)experiment. Transwell assay was used to analyze cell migration and invasion abilities. Dual-luciferase reporter assay andRNA Immunoprecipitation (RIP) assay were performed to confirm the targeted interaction between miR-195 and SOX4.Our data supported that miR-195 was downregulated and SOX4 was upregulated in EC tissues and cell lines. Upregulationof miR-195 inhibited migration, invasion and epithelial-mesenchymal transition (EMT) of EC cells. Moreover, SOX4 was adirect target of miR-195. MiR-195 overexpression-mediated anti-migration, anti-invasion and anti-EMT effects wereantagonized by SOX4 restoration in EC cells. In conclusion, our study suggested that miR-195 inhibited the migration,invasion and epithelial mesenchymal transition (EMT) of EC cells at least partly by targeting SOX4. Our study provided anovel underlying mechanism for EC metastasis and a promising therapeutic target for EC management.

Long Non-Coding RNA TUG1 Promotes Proliferation and Inhibits Apoptosis of Osteosarcoma Cells by Sponging miR-132-3p and Upregulating SOX4 Expression

PURPOSE: Long non-coding RNA taurine upregulated gene 1 (TUG1) is reported to be a vital regulator of the progression of various cancers. This study aimed to explore the exact roles and molecular mechanisms of TUG1 in osteosarcoma (OS) development. MATERIALS AND METHODS: Real-time quantitative PCR was applied to detect the expressions of TUG1 and microRNA-132-3p (miR-132-3p) in OS tissues and cells. Western blot was performed to measure protein levels of sex determining region Y-box 4 (SOX4). Cell viability was assessed using XTT assay. Cell apoptosis was evaluated using flow cytometry and caspase-3 activity detection assays. Bioinformatics analysis and luciferase reporter experiments were employed to confirm relationships among TUG1, miR-132-3p, and SOX4. RESULTS: TUG1 was highly expressed in human OS tissues, OS cell lines, and primary OS cells. TUG1 knockdown hindered proliferation and induced apoptosis in human OS cell lines and primary OS cells. Moreover, TUG1 inhibited miR-132-3p expression by direct interaction, and introduction of miR-132-3p inhibitor partly abrogated the effect of TUG1 knockdown on the proliferation and apoptosis of OS cells. Furthermore, SOX4 was validated as a target of miR-132-3p. Further functional analyses revealed that miR-132-3p inhibited proliferation and induced apoptosis of OS cells, while this effect was greatly abated following SOX4 overexpression. Moreover, TUG1 knockdown suppressed proliferation and promoted apoptosis by upregulating miR-132-3p and downregulating SOX4 in primary OS cells. CONCLUSION: TUG1 facilitated proliferation and suppressed apoptosis by regulating the miR-132-3p/SOX4 axis in human OS cell lines and primary OS cells. This finding provides a potential target for OS therapy.

Propofol inhibits proliferation, migration, and invasion but promotes apoptosis by regulation of Sox4 in endometrial cancer cells

Propofol is an intravenous sedative hypnotic agent of which the growth-inhibitory effect has been reported on various cancers. However, the roles of propofol in endometrial cancer (EC) remain unclear. This study aimed to explore the effects of propofol on EC in vitro and in vivo. Different concentrations of propofol were used to treat Ishikawa cells. Colony number, cell viability, cell cycle, apoptosis, migration, and invasion were analyzed by colony formation, MTT, flow cytometry, and Transwell assays. In addition, the pcDNA3.1-Sox4 and Sox4 siRNA plasmids were transfected into Ishikawa cells to explore the relationship between propofol and Sox4 in EC cell proliferation. Tumor weight in vivo was measured by xenograft tumor model assay. Protein levels of cell cycle-related factors, apoptosis-related factors, matrix metalloproteinases 9 (MMP9), matrix metalloproteinases 2 (MMP2) and Wnt/β-catenin pathway were examined by western blot. Results showed that propofol significantly decreased colony numbers, inhibited cell viability, migration, and invasion but promoted apoptosis in a dose-dependent manner in Ishikawa cells. Moreover, propofol reduced the expression of Sox4 in a dose-dependent manner. Additionally, propofol significantly suppressed the proportions of Ki67+ cells, but Sox4 overexpression reversed the results. Furthermore, in vivo assay results showed that propofol inhibited tumor growth; however, the inhibitory effect was abolished by Sox4 overexpression. Moreover, propofol inhibited Sox4 expression via inactivation of Wnt/β-catenin signal pathway. Our study demonstrated that propofol inhibited cell proliferation, migration, and invasion but promoted apoptosis by regulation of Sox4 in EC cells. These findings might indicate a novel treatment strategy for EC.

SOX4 gene and lung development / 国际儿科学杂志

SOX4 is a member of the group C subfamily of the SOX transcription factors and has a criti-cal role during embryogenesis and in controlling cell fate.SOX4 can participate in TGF-β,Wnt signaling ways to mediate cell differentiation,proliferation,migration.SOX4 can facilitate epithelial-mesenchymal transition by di-rectly regulating EZH2 expression.It is one of the 64 genes that constitute a general signature in all human canc-ers.SOX4 is widely expressed in lung and establishes the lung structure in lung development.Here,we provide an overview of role of SOX4 in cell and lung development.

Expression of SOX4 and EZH2 in breast cancer and their clinicopathologic significance / 临床与实验病理学杂志

Purpose To explore the expression of two epithelial-mesenchymal transition (EMT) regulators,Sry-related high mobility group box 4 (SOX4) and enhancer of zeste homolog 2 (EZH2),in breast cancer tissues and their clinicopathologic significances.Methods The expression of SOX4 and EZH2 was detected by immunohistochemistry of EnVision twostep in paraffin-embedded tissue sections from 241 cases of breast cancer and 126 cases of paracancerous breast tissues.The relationship between their expression and clinicopathologic characteristics was statistically analyzed,and their correlation was also analyzed.Results In breast cancer tissues and paracancerous breast tissues,the high expression rates of SOX4 were 82.2％ and 57.1％ respectively,while the high expression rates of EZH2 were 80.1％ and 47.6％ respectively.Their expression in breast cancer tissues was higher than that in paracancerous breast tissues (P ＜ 0.05).High SOX4 expression was correlated with lymph node metastasis,high TNM stage,and overexpression of HER-2 (P ＜ 0.05),while high EZH2 expression was correlated with high histological grade,lymph node metastasis,ER negativity,EGFR positivity and high poliferation index of Ki67 (P ＜ 0.05).There was a positive correlation between SOX4 and EZH2 expression in breast cancer tissues (rs =0.256,P ＜0.001).Conclusion EMT regulators SOX4 and EZH2 might be responsible for the metastasis of breast cancer,which are upregulated in breast cancer tissues.They may act as potential biomarkers to predict the metastasis of breast cancer.

Repressive effect of miRNA-363 via SOX4 on human osteosarcoma cell growth and apoptosis / 中国病理生理杂志

AIM:To detect the expression of miRNA-363 and SOX4 in osteosarcoma tissues and to investigate the effect of miRNA-363 on the viability and apoptosis of human osteosarcoma cell line MG-63.METHODS: Real-time PCR was used to detect the expression level and the relationship of miRNA-363 and SOX4 mRNA in the osteosarcoma tis-sues and the corresponding paratumorous tissues collected from 63 patients.The expression levels of miRNA-363 and SOX4 in osteosarcoma cell line MG-63 after transfected with miRNA-363 mimics were measured .The cell viability was measured by CCK-8 assay.Flow cytometry was used to monitor the changes of cell cycle and apoptosis .The changes of SOX4 and miRNA-363 expression levels in the MG-63 cells after transfection with SOX4 siRNA or pcDNA/SOX4 was detect by real-time PCR.RESULTS:The expressed level of miRNA-363 was lower , and the expression level of SOX 4 was higher in the osteosarcoma tissues than those in the adjacent normal tissues .A significantly negative correlation between the expression levels of miRNA-363 and SOX4 was observed .The expression of miRNA-363 in the MG-63 cells after transfection with miRNA-363 mimics was significantly up-regulated, while the expression of SOX4 in the MG-63 cells was significantly down-regulated , with significant difference as compared with the cells transfected with miRNA-NC and control cells .The viability of MG-63 cells was inhibited, the cell cycle was arrested in G0/G1 phase, and the cell apoptosis was increased by transfec-tion with miRNA-363 mimics.The relative protein expression levels of SOX 4 in SOX4 siRNA group and pcNDA/SOX4 group were significantly different from those in negative control group , but the relative expression levels of miRNA-363 had no significant difference .Over-expression of SOX4 restored the viability of the MG-63 cells reduced by miR-363.CON-CLUSION:The expression level of miRNA-363 is low in human osteosarcoma tissue .miRNA-363 may inhibits the viabili-ty of osteosarcoma cell line MG-63 and promotes cell apoptosis in vitro via inhibiting the SOX4 expression.

Expressions and roles of SOX families in glioma / 国际肿瘤学杂志

Glioma is a common malignant tumor of nervous system.Many genes of SOX family are closely related to the genesis and development of glioma,among which SOX2,SOX4,SOX7,SOX9 gene are all abnormal in glioma.SOX2 gene is highly expressed in glioma and has positive correlation with the malignant grade of glioma.SOX2 gene coordinates with many factors and promotes the genesis and development of glioma.SOX4 gene plays as both a oncogene and a tumor suppressor gene in brain glioma,and can regulate a variety of factors.SOX7,as a tumor suppressor gene,shows low expression in glioma,and suppresses the genesis of brain glioma by regulating the related factors and signaling pathways.SOX9 gene is highly expressed in glioma tissues,and promotes tumorigenesis and metastasis of glioma by coordinating with a variety of factors,and can be used as an important risk factor for the prognosis of glioma patients.

Expressions and roles of SOX families in glioma / 国际肿瘤学杂志

Glioma is a common malignant tumor of nervous system.Many genes of SOX family are closely related to the genesis and development of glioma,among which SOX2,SOX4,SOX7,SOX9 gene are all abnormal in glioma.SOX2 gene is highly expressed in glioma and has positive correlation with the malignant grade of glioma.SOX2 gene coordinates with many factors and promotes the genesis and development of glioma.SOX4 gene plays as both a oncogene and a tumor suppressor gene in brain glioma,and can regulate a variety of factors.SOX7,as a tumor suppressor gene,shows low expression in glioma,and suppresses the genesis of brain glioma by regulating the related factors and signaling pathways.SOX9 gene is highly expressed in glioma tissues,and promotes tumorigenesis and metastasis of glioma by coordinating with a variety of factors,and can be used as an important risk factor for the prognosis of glioma patients.

Expression of transcription factors SOX4 and C/EBPα in chronic myeloid leukemia and their significances / 白血病·淋巴瘤

Objective To investigate the expression of SOX4 and C/EBPα mRNA in chronic myeloid leukemia (CML) and their clinical significances. Methods Bone marrow samples from 68 cases of CML including 57 newly diagnosed patients and 11 patients treated with imatinib were collected, and peripheral blood mononuclear cells from 30 healthy people were collected as healthy control. The expression of SOX4 and C/EBPαmRNA and protein levels were detected by RT-PCR and Western blot, respectively. The relations between the expression of SOX4 and C/EBPα and the influences of imatinib on SOX4 and C/EBPα were analyzed. Results The expression level of SOX4 mRNA was increased in newly diagnosed CML patients compared with that of normal control group (6.545 5±1.495 2 vs. 0.059 6±0.018 8, t=3.139, P=0.002 3), but the expression level of C/EBPαmRNA was significantly decreased (0.238 8±0.033 8 vs. 0.810 5±0.056 2, t=9.240, P0.05). The expression level of SOX4 mRNA in 5 patients treated with imatinib was decreased (0.120 6 ±0.044 9 vs. 0.557 9±0.144 8, t=2.885, P=0.020 4), and the expression level of C/EBPαmRNA was increased (0.330 3±0.042 4 vs. 0.150 5±0.046 5, t=2.855, P=0.021 3). The expression level of SOX4 mRNA in 6 patients who developed blast phase during the treatment of imatinib was increased (0.469 9±0.123 0 vs. 0.050 2±0.036 6, t=2.370, P=0.039 3), and the expression level of C/EBPα mRNA was decreased (0.197 9 ±0.064 7 vs. 0.378 7±0.042 9, t=2.327, P=0.042 3). The expression of SOX4 mRNA was negatively correlated with C/EBPα mRNA (t=-0.554 6, P=0.002 8). Conclusions In newly diagnosed CML, the expression level of SOX4 is increased, C/EBPα is decreased compared with that of healthy control, and both have negative correlation. In the patients in blast phase after imatinib treatment, SOX4 gene is up-regulated, and C/EBPα is down-regulated. C/EBPα-SOX4 axis may play a role in the occurrence and development of CML. SOX4 may be a new molecular target for the treatment of CML.

Over-expression of Sox4 and β-catenin is associated with a less favorable prognosis of osteosarcoma / 华中科技大学学报（医学）（英德文版）

The purpose of this study was to examine the association of the expression of Sox4 and β-catenin with the prognosis of osteosarcoma. A total of 108 cases of conventional osteosarcoma were involved in this study and 28 cases of osteochondroma served as controls. The expression of Sox4 and β-catenin was detected by using immunohistochemical staining and Western blotting. The results showed that Sox4 and β-catenin were over-expressed in 67 (62.03%) and 62 (57.41%) of 108 osteosarcoma cases, while in only 3 (10.71%) and 5 (17.86%) of 28 controls, respectively (P<0.05 for all). The expression of Sox4 and β-catenin was associated with the distant metastasis, pathological grade and Enneking stage of patients with osteosarcoma (P<0.05 for all). The mean overall survival time and the 5-year-survival rate in osteosarcoma patients with Sox4 and β-catenin over-expressed were significantly reduced as compared with those in Sox4 and β-catenin low-expression group (P<0.05 for all). Cox multifactor regression analysis revealed that the distant metastasis, Enneking stage, and the expression of Sox4 and β-catenin were independent risk factors of patients with osteosarcoma (P<0.05 for all). The findings indicated that overexpression of Sox4 and β-catenin is associated with a poor prognosis of osteosarcoma.

Progress of SOX4 in leukemia / 国际儿科学杂志

Leukemia is a polygenic malignant proliferative disease in the hematopoietic system.Its possible causative genes and gene therapy are one hotspot of the current researches.Sex-determining region Y-related high-mobility-group box transcription factor 4(SOX4) is a member of the group C subfamily of the SOX transcription factors,playing a critical role in the occurrence and development of many tumors such as lung cancer,breast cancer,hepatic carcinoma,gastric cancer and medulloblastoma.Recently,researches have found that SOX4 is significant in the occurrence,treatment and prognosis evaluation of leukemia.This paper reviews the structure and functions of SOX4,its expression and mechanism in leukemia,and its influence on leukemia treatment and prognosis.

Effect and Mechanism of Ouabain in Regulating Proliferation of Human Esophageal Carcinoma Cells / 胃肠病学

Background:Esophageal cancer is a common gastrointestinal cancer with poor prognosis,and effective chemotherapy is lacking currently. Studies have shown that cardiac glycosides can inhibit tumor cells growth,but its mechanism has not been fully clarified. Aims:To investigate the effect and mechanism of ouabain in regulating proliferation of human esophageal carcinoma cells. Methods:OE19 human esophageal carcinoma cells were treated with ouabain,and cells in control group were treated with DMSO. Cell proliferation was assessed by cell counting method. mRNA expressions of Sox2,Sox4,Sox7,Sox9 and Sox10 were determined by real-time PCR. Protein expression of Sox4 was determined by Western blotting. Gene expressions of phospho-histone3( ph3),a cell proliferation marker and Sox4 were detected by immunofluorescence staining. Results:Ouabain( ≥ 40 nmol/ L)could significantly inhibit OE19 cells proliferation. mRNA and protein expressions of Sox4 were significantly decreased in OE19 cells in ouabain(40 nmol/ L)group than those in control group(P 0. 05). Gene expressions of ph3 and Sox4 in nucleus of OE19 cells were decreased in ouabain (40 nmol/ L)group than those in control group. Conclusions:Ouabain is effective in inhibiting human esophageal carcinoma cells proliferation,the underlying mechanism might be related with down-regulation of Sox4 expression and the subsequent cell cycle modulation.

The identification of human microRNA335′s predicated target gene CCL1 1,CCL26 and SOX4 / 重庆医学

Objective To identify the targeted-regulating relationship between human MicroRNA335 (hsa-miR-335 )and CCL1 1,CCL26 and SOX4.Methods The potential fragments of hsa-miR-335 target genes CCL1 1,CCL26 and SOX4 were predicted by the bioinformatics analyzing tools online.The 3′untranslated regions(3′UTR)of the CCL1 1,CCL26 and SOX4 were connected to the eukaryotic expression vectors pMIR REPORT.The constructs of pMIR-REPORT-CCL1 13′UTR,pMIR-REPORT-CCL26 3′UTR,pMIR-REPORT-SOX4 3′UTR and positive control were co-transfected with Pre-miRTM miRNA335 Precursor or negative control into 293 T7/1 7 cell line by lipofectamine 2000,respectively.Both Firefly and Renilla luciferase activity were detected by dual luciferase reporter assay system.Results Compared with the negative control group,luciferase assay revealed that has-miR-335 could significantly diminish luciferase activity from SOX4 reporter vector (P 0.05).Conclusion The results suggested that hsa-miR-335 targeted regu-lated SOX4,but not targeted CCL1 1 and CCL26.

Research progress of the abnormal expression of Sox genes in breast cancer / 国际外科学杂志

Sox gene family is composed of a class of SRY gene,encoding a series of transcription factors.In the ontogeny,sox genes involved in a variety of development,such as sex determination and differentiation,neural development,cartilage formation.In recent years,researchers found that the abnormal expression of sox gene was related with the development of breast cancer,such as,the overexpression of Sox2 and Sox4 was related with breast cancer; Sox7 and Sox17 in breast cancer could act as tumor suppressor gene,the downregulation of which could activate Wnt/β-catenin signaling pathway.

Mechanism and abnormal expression of sox4 gene in tumor / 国际肿瘤学杂志

Recent studies have revealed that sox4 gene expresses abnormally in many kinds of tumor tissues and it probably involves in tumorigenesis,development and metastasis of cancer.Regulating proliferation,differentiation,apoptosis and other process may participate in the work mechanisms of sox4 gene.Therefore,further studies about the relationship between sox4 gene and tumor would provide new ideas of exploring special diagnosis markers and novel targets for tumor therapy.

Inhibition of Melanoma Cell Proliferation by Targeting Wnt/β-catenin Pathway through Sox4 RNA Interference / 华中科技大学学报（医学）（英德文版）

The effect of siRNA-mediated Sox4 gene silencing on Wnt/β-catenin signaling pathway of human malignant melanoma cell line A375 was investigated.Two types of dsRNA targeting Sox4 were constructed and transfected into A375 cells,and untreated cells and cells transfected with scramble RNA were used as blank control and negative control respectively.The expression levels of mRNA and protein of Sox4,Wnt3a,β-catenin and Wnt/β-catenin signaling target gene Survivin were detected after real-time PCR and Western blot respectively.MTT assay was used to measure cell proliferation after Sox4 knockdown.β-catenin/TCF transcription reporter assay was used for assessing Wnt/β-catenin signaling pathway activity.Our results showed that the two types of Sox4 siRNA were transfected into A375 cells successfully.As compared with untreated cells,Sox4 siRNAs had no significant influence on Wnt3a expression,and Sox4 siRNAs led to the decrease of β-catenin at protein level.Wnt/β-catenin signaling pathway activity was inhibited significantly.As a target of Wnt/β-catenin signaling,Survivin was decreased at both mRNA and protein levels,and cell proliferation was attenuated.Our study suggests that Sox4 may play an important role in Wnt/β-catenin signaling pathway in human malignant melanoma cells by regulating β-catenin protein level,indicating that Sox4 is involved in the progression of malignant melanoma thromgh Wnt/β-catenin signaling pathway.

Role of menin in regulation of SOX4 gene expression / 上海第二医科大学学报

Objective To explore the role of menin in regulation of SOX4 gene expression. Methods Reporter gene vectors with SOX4 promoter were constructed,and the influence of menin on SOX4 gene promoter activity was analyzed by dual-luciferase reporter gene assay system.Real-time PCR was employed to detect the expression of SOX4 gene in mef cells of MEN1-/-mice and wild-type mef cells. Results Compared with wild-type mef cells,the SOX4 gene promoter activity was significantly higher in mef cells of MEN1-/-mice.After MEN1 gene was transfected into mef cells of MEN1-/-mice,the SOX4 gene promoter activity significantly descreased.The expression of SOX4 gene in mef cells of MEN1-/-mice was 2.5 time higher than that in wild-type mef cells. Conclusion Menin can inhibit the expression of SOX4 gene.

Involvement of Sox-4 in the cytochrome c-dependent AIF-independent apoptotic pathway in HeLa cells induced by delta12-prostaglandin J2

delta12-Prostaglandin (PG) J2 is known to elicit an anti-neoplastic effects via apoptosis induction. Previous study showed delta12-PGJ2-induced apoptosis utilized caspase cascade through cytochrome c-dependent pathways in HeLa cells. In this study, the cellular mechanism of delta12-PGJ2- induced apoptosis in HeLa cells, specifically, the role of two mitochondrial factors; bcl-2 and apoptosis-inducing factor (AIF) was investigated. Bcl-2 attenuated delta12-PGJ2-induced caspase activation, loss of mitochondrial transmembrane potential (delta psi m), nuclear fragmentation, DNA laddering, and growth curve inhibition for approximately 24 h, but not for longer time. AIF was not released from mitochondria, even if the delta psi m was dissipated. One of the earliest events observed in delta12-PGJ2-induced apoptotic events was dissipation of delta psi m, the process known to be inhibited by bcl-2. Pre-treatment of z-VAD- fmk, the pan-caspase inhibitor, resulted in the attenuation of delta psi m depolarization in delta12-PGJ2- induced apoptosis. Up-regulation of Sox-4 protein by delta12-PGJ2 was observed in HeLa and bcl-2 overexpressing HeLa B4 cell lines. Bcl-2 overexpression did not attenuate the expression of Sox-4 and its expression coincided with other apoptotic events. These results suggest that delta12-PGJ2 induced Sox-4 expression may activate another upstream caspases excluding the caspase 9-caspase 3 cascade of mitochondrial pathway. These and previous findings together suggest that delta12-PGJ2-induced apoptosis in HeLa cells is caspase-dependent, AIF-independent events which may be affected by Sox-4 protein expression up-regulated by delta12-PGJ2.

Sox-4 is a positive regulator of Hep3B and HepG2 cells' apoptosis induced by prostaglandin (PG)A2 and 12-PGJ2

We reported earlier that expression of Sox-4 was found to be elevated during prostaglandin (PG) A2 and delta(12)-PGJ(12) induced apoptosis in human hepatocarcinoma Hep3B cells. In this study, the role of Sox-4 was examined using human Hep3B and HepG2 cell lines. Sox-4 induction by several apoptotic inducer such as A23187 (Ca(2+) ionophore) and etoposide (topoisomerase II inhibitor) and Sox-4 transfection into the cells were able to induce apoptosis as observed by the cellular DNA fragmentation. Antisense oligonucleotide of Sox-4 inhibited the induction of Sox-4 expression and blocked the formation of DNA fragmentation by PGA(2) and delta(12)-PGJ(12) in Hep3B and HepG2 cells. Sox-4-induced apoptosis was accompanied with caspase-1 activation indicating that caspase cascade was involved in this apoptotic pathway. These results indicate that Sox-4 is involved in Hep3B and HepG2 cells apoptosis as an important apoptotic mediator.