Neural and Neural Crest-Derived Stem Cell Distribution in the Adult Human Dental Pulp

SUMMARY: Despite comprehensive studies and reports about the properties of dental pulp stem cells (DPSCs) in vitro, we still need to confirm whether these in vitro characteristics coincide with the nature of DPSCs in situ. The anatomical location of DPSCs populations in the dental pulp has yet to be investigated. Moreover, the mesenchymal DPSCs have been much more studied than the neural crest-derived DPSCs. In this study, well-recognized neural/neural crest stem cell markers NCAM1, Nestin, SNAIL/SLUG, SOX9, and S100 are being investigated by immunohistochemistry to localize the precise location of these populations of DPSCs within the human adult dental pulp.All previously mentioned markers were expressed in the dental pulp, and their intensity and location of expression were reported.

A pesar de estudios e informes exhaustivos sobre las propiedades de las células madre de la pulpa dental (DPSC) in vitro, todavía necesitamos confirmar si estas características in vitro coinciden con la naturaleza de las DPSC in situ. La ubicación anatómica de las poblaciones de DPSC en la pulpa dental aún no se ha investigado. Además, las DPSC mesenquimales han sido mucho más estudiadas que las DPSC derivadas de la cresta neural. En este estudio, se están investigando mediante inmunohisto química marcadores de células madre de la cresta neural/ neural NCAM1, Nestin, SNAIL/SLUG, SOX9 y S100 para localizar la ubicación precisa de estas poblaciones de DPSC dentro de la pulpa dental humana adulta. Todos los marcadores mencionados anteriormente se expresaron en la pulpa dental y se informó su intensidad y ubicación de expresión.

Circ_0075825 promotes gastric cancer progression via adsorbing miR-432-5p to modulate SOX9

Abstract Methods Circ_0075825 expression in adjacent tissues and GC tissues was evaluated by bioinformatics method and quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR). How circ_0075825 regulated GC cell growth, migration, invasion, and apoptosis were investigated by cell counting kit-8 assay, transwell assay and flow cytometry. The targeted interplays among circ_0075825, and miR-432-5p and Sex-Determining Region Y-related high-mobility group box 9 (SOX9) were explored by bioinformatics analysis and luciferase reporter gene assay. The regulatory effects of circ_0075825 and miR-432-5p on SOX9 protein expression were probed by western blot. Results Circ_0075825 expression was raised in GC tissues and cell lines. Circ_0075825 overexpression promoted the proliferative, migrative and invasive abilities of GC cells, while inhibiting apoptosis, while depletion of circ_0075825 suppressed the malignant biological behaviors of GC cells. SOX9 was identified as one of the direct target genes of miR-432-5p, and circ_0075825 repressed the expression of miR-432-5p, to induce the expression of SOX9. Furthermore, miR-432-5p overexpression counteracted the promoting effect of circ_0075825 on the malignancy of GC cells. Conclusion Circ_0075825 promotes GC progression via sponging miR-432-5p to regulate SOX9 expression level, and it may be a novel therapeutic target for treating GC.

Regulatory role of the transcriptional coactivator Mediator 1 in skin hair regeneration and its mechanisms / 中华皮肤科杂志

Objective:To investigate the effect of the transcriptional coactivator Mediator 1 (Med1) on mouse hair regeneration, and to explore potential mechanisms.Methods:Med1 flox/flox C57BL/6J mice were mated with K14-Cre mice, and the mice with epidermis-specific knockout of Med1 gene, namely K14-Cre-expressing Med1 flox/flox mice (knockout group) , were obtained by using the Cre-Loxp system, while Med1 flox/flox mice without K14-Cre expression served as control group. Mice in the two groups (3 mice in each group) were raised together for 8 weeks followed by dorsal hair removal. Hair regeneration was observed for 12 consecutive days after hair removal. After 12 days, all mice in the two groups were sacrificed, their depilated and non-depilated dorsal skin tissues were resected, and total RNA was extracted from the tissues. Real-time quantitative PCR was performed to determine the mRNA expression of hair keratin genes, vitamin D receptor/β-catenin pathway-related genes, and genes associated with maintenance of hair follicle stem cell proliferation and quiescence. Paraffin-embedded sections of depilated and non-depilated mouse skin tissues were prepared, and immunofluorescence staining was conducted to determine the number of stem cells in the hair follicle bulge. Two-independent-sample t test was used for comparisons between two groups. Results:From days 0 to 12 after depilation, hair regeneration was delayed in the depilated skin area in the knockout group compared with the control group. Real-time quantitative PCR showed significantly decreased mRNA relative expression levels of hair keratin genes Ha1 and Krt2-16, vitamin D receptor/β-catenin pathway-related genes S100a3, Dlx3 and Tubb3, and genes associated with maintenance of hair follicle stem cell proliferation and quiescence including Lhx2, Sox9 and Nfatc1 in the depilated skin tissues in the knockout group (22.09 ± 12.32, 2.07 ± 0.20, 0.02 ± 0.01, 12.36 ± 2.12, 1.75 ± 0.46, 0.39 ± 0.02, 4.42 ± 0.76, 0.44 ± 0.07, respectively) compared with the control group (70.53 ± 9.46, 7.76 ± 0.49, 0.05 ± 0.01, 26.16 ± 2.96, 2.60 ± 0.14, 0.71 ± 0.09, 11.93 ± 0.42, 0.75 ± 0.04, respectively; t = 5.40, 18.64, 3.89, 6.57, 3.04, 6.10, 15.03, 6.18, respectively, all P < 0.05) . Immunofluorescence staining showed that the number of CD34 +K15 + hair follicle stem cells in the hair follicle bulge in both depilated and non-depilated skin tissues was significantly lower in the knockout group than in the control group. Conclusion:Med1 gene knockout may down-regulate the expression of downstream genes of the vitamin D receptor/β-catenin pathway and genes associated with maintenance of hair follicle stem cell proliferation and quiescence (Sox9, Nfatc1 and Lhx2) , and reduce the number of hair follicle stem cells, leading to hair follicle differentiation disorder and hair regeneration delay.

Correlation between the expressions of miR-524-5p and SOX9 in advanced gastric cancer and their influences on chemotherapy efficacy and prognosis / 国际肿瘤学杂志

Objective:To investigate the correlation between the expressions of miR-524-5p and sex determining region Y box protein 9 (SOX9) in advanced gastric cancer and their influences on the efficacy and prognosis of chemotherapy.Methods:A total of 82 patients diagnosed as advanced gastric cancer who received DCF (docetaxel + cisplatin + fluorouracil) chemotherapy in 910th Hospital of Joint Logistics Support Force of Chinese People′s Liberation Army from January 2015 to December 2017 were selected as the research objects. The expression levels of miR-524-5p and SOX9 in gastric cancer tissues were detected by fluorescence quantitative PCR. The correlation between the expressions of miR-524-5p and SOX9 was analyzed, the total effective rates of chemotherapy in patients with different miR-524-5p and SOX9 expression levels were compared, and the predictive value of miR-524-5p combined with SOX9 on the efficacy of chemotherapy was analyzed. Overall survival (OS) and progression-free survival (PFS) were analyzed.Results:The expressions of miR-524-5p and SOX9 were not related to gender or age of patients with advanced gastric cancer (all P>0.05), but were related to the degree of differentiation ( χ2=3.577, P=0.001; χ2=5.654, P<0.001) and distant metastasis ( χ2=2.466, P=0.016; χ2=5.218, P<0.001) of patients with advanced gastric cancer. There was a negative correlation between the expressions of miR-524-5p and SOX9 in advanced gastric cancer ( r=-0.348, P=0.001). According to the median expressions of miR-524-5p and SOX9 in gastric cancer tissues, patients were divided into high expression and low expression groups, miR-524-5p≥0.64 was high expression ( n=41), <0.64 was low expression ( n=41), SOX9≥1.84 was high expression ( n=41), and <1.84 was low expression ( n=41). The total effective rate of advanced gastric cancer patients with high expression of miR-524-5p was 58.54% (24/41), which was higher than that of patients with low expression of miR-524-5p (24.39%, 10/41), and there was a statistically significant difference ( χ2=9.484, P=0.002). The total effective rate of advanced gastric cancer patients with high expression of SOX9 was 21.95% (9/41), which was lower than 60.97% (25/41) of patients with low expression of SOX9, and there was a statistically significant difference ( χ2=12.863, P<0.001). Receiver operating characteristic curve analysis showed that the expressions of miR-524-5p and SOX9 had predictive value for the efficacy of chemotherapy, and the area under the curve was 0.753 (95% CI: 0.644-0.861, P<0.001) and 0.660 (95% CI: 0.540-0.780, P=0.014) respectively. The combination of miR-524-5p and SOX9 had predictive value for the efficacy of chemotherapy, and the area under the curve was 0.768 (95% CI: 0.667-0.868, P<0.001). The median PFS and OS of patients with high expression of miR-524-5p were 8 months and 14 months, which were longer than those of patients with low expression of miR-524-5p (6 months, 9 months), and there were statistically significant differences ( χ2=21.160, P<0.001; χ2=29.730, P<0.001). The median PFS and OS of patients with high expression of SOX9 were 7 months and 10 months, which were shorter than those of patients with low expression of SOX9 (8 months, 12 months), and there were statistically significant differences ( χ2=6.345, P=0.012; χ2=4.107, P=0.043). Conclusion:There is a negative correlation between the expressions of miR-524-5p and SOX9 in advanced gastric cancer tissues. The chemotherapy efficacy and prognosis of patients with high expression of miR-524-5p and low expression of SOX9 are better than those of patients with low expression of miR-524-5p and high expression of SOX9.

Effect of pomegranate peel polyphenols on sebaceous gland spots and AKT/Sox9 signaling pathway in golden hamsters / 中华皮肤科杂志

Objective:To evaluate the effect of pomegranate peel polyphenols on sebum secretion in golden hamsters, and to explore its possible mechanisms.Methods:Thirty golden hamsters were randomly and equally divided into 5 groups: (ointment) vehicle group, 0.48%-, 0.96%-, 1.92%-pomegranate peel polyphenol ointment groups, and retinoic acid cream group. Corresponding cream or ointments were applied to bilateral sebaceous gland spots of the golden hamsters at a dose of 1 gram twice a day for 4 consecutive weeks. The area of bilateral sebaceous gland spots was measured on days 0, 7, 14, 21 and 28 after the start of treatment, which was calculated by the maximum longitudinal diameter multiplied by the maximum transverse diameter. Twenty-four hours after the last treatment, immunohistochemical study was conducted to determine the expression of AKT/Sox9 signaling pathway in sebaceous gland spots resected from the golden hamsters. The area of sebaceous gland spots in these groups at different time points was compared by repeated measures analysis of variance, and other data were analyzed by one-way analysis of variance or Kruskal-Wallis H test. Results:The area of sebaceous gland spots was significantly smaller in the 0.96%-, 1.92%-pomegranate peel polyphenol ointment groups (50.48±2.41 mm 2, 48.24±2.56 mm 2, respectively) and retinoic acid cream group (48.31±2.76 mm 2) than in the vehicle group (57.99±3.29 mm 2; t=2.69, 3.98, 3.65, P=0.012, 0.001, 0.001, respectively) . Sox9 expression was significantly lower in the 1.92%-pomegranate peel polyphenol ointment group (0.39±0.04) and retinoic acid cream group (0.38±0.03) than in the vehicle group (0.44±0.02, P=0.040) . However, there was no significant difference in AKT expression among the 5 groups ( F=1.645, P=0.199) . Conclusion:Pomegranate peel polyphenols can reduce the sebaceous gland spot area and inhibit sebum secretion in golden hamsters, which may be related to the inhibition of Sox9 expression.

microRNA-130b downregulation potentiates chondrogenic differentiation of bone marrow mesenchymal stem cells by targeting SOX9

Osteoarthritis (OA) is a chronic health condition. MicroRNAs (miRs) are critical in chondrocyte apoptosis in OA. We aimed to investigate the mechanism of miR-130b in OA progression. Bone marrow mesenchymal stem cells (BMSCs) and chondrocytes were first extracted. Chondrogenic differentiation of BMSCs was carried out and verified. Chondrocytes were stimulated with interleukin (IL)-1β to imitate OA condition in vitro. The effect of miR-130b on the viability, inflammation, apoptosis, and extracellular matrix of OA chondrocytes was studied. The target gene of miR-130b was predicted and verified. Rescue experiments were performed to further study the underlying downstream mechanism of miR-130b in OA. miR-130b first increased and drastically reduced during chondrogenic differentiation of BMSCs and in OA chondrocytes, respectively, while IL-1β stimulation resulted in increased miR-130b expression in chondrocytes. miR-130b inhibitor promoted chondrogenic differentiation of BMSCs and chondrocyte growth and inhibited the levels of inflammatory factors. miR-130b targeted SOX9. Overexpression of SOX9 facilitated BMSC chondrogenic differentiation and chondrocyte growth, while siRNA-SOX9 contributed to the opposite trends. Silencing of SOX9 significantly attenuated the pro-chondrogenic effects of miR-130b inhibitor on BMSCs. Overall, miR-130b inhibitor induced chondrogenic differentiation of BMSCs and chondrocyte growth by targeting SOX9.

microRNA-130b downregulation potentiates chondrogenic differentiation of bone marrow mesenchymal stem cells by targeting SOX9

Osteoarthritis (OA) is a chronic health condition. MicroRNAs (miRs) are critical in chondrocyte apoptosis in OA. We aimed to investigate the mechanism of miR-130b in OA progression. Bone marrow mesenchymal stem cells (BMSCs) and chondrocytes were first extracted. Chondrogenic differentiation of BMSCs was carried out and verified. Chondrocytes were stimulated with interleukin (IL)-1β to imitate OA condition in vitro. The effect of miR-130b on the viability, inflammation, apoptosis, and extracellular matrix of OA chondrocytes was studied. The target gene of miR-130b was predicted and verified. Rescue experiments were performed to further study the underlying downstream mechanism of miR-130b in OA. miR-130b first increased and drastically reduced during chondrogenic differentiation of BMSCs and in OA chondrocytes, respectively, while IL-1β stimulation resulted in increased miR-130b expression in chondrocytes. miR-130b inhibitor promoted chondrogenic differentiation of BMSCs and chondrocyte growth and inhibited the levels of inflammatory factors. miR-130b targeted SOX9. Overexpression of SOX9 facilitated BMSC chondrogenic differentiation and chondrocyte growth, while siRNA-SOX9 contributed to the opposite trends. Silencing of SOX9 significantly attenuated the pro-chondrogenic effects of miR-130b inhibitor on BMSCs. Overall, miR-130b inhibitor induced chondrogenic differentiation of BMSCs and chondrocyte growth by targeting SOX9.

Differences in the effect of two injectable Pluronic F-127 hydrogel composites in repairing rabbit cartilage defects / 中国组织工程研究

BACKGROUND: Preliminary experiments show that bone marrow mesenchymal stem cells transfected with SOX9 gene can grow and proliferate in Pluronic F-127 hydrogel, promote the secretion of extracellular matrix, and increase the expression of cartilage matrix. OBJECTIVE: The SOX9 gene was transduced into bone marrow mesenchymal stem cells by lentivirus gene induction, and then combined with injectable Pluronic F-127 hydrogel to observe the effect of Pluronic F-127 hydrogel on repairing cartilage defects. METHODS: SOX9 gene was transfected into bone marrow mesenchymal stem cells by lentivirus gene induction. After 48 hours of transfectlon, SOX9 gene was combined with Pluronic F-127 hydrogel. Sixty New Zealand white rabbits provided by the Experimental Animal Center of Wuhan University of Science and Technology were selected to establish the models of femoral condylar cartilage defect of the right knee joint. The rabbits were randomly divided into three groups: model group without implantation of any material at the defect site, control group with implantation of non-transfected bone marrow mesenchymal stem cells and Pluronic F-127 hydrogel complex at the defect site, and experimental group with implantation of SOX9 gene-transfected bone marrow mesenchymal stem cells and Pluronic F-127 hydrogel complex at the defect site. Four and twelve weeks after operation, the defect tissues were taken for three-dimensional reconstruction of micro-CT, hematoxylin-eosin staining, Safranine O staining, type II collagen immunohistochemical staining and Wakitani soft tissue repair histological score. This study was approved by the Ethics Committee of Wuhan University of Science and Technology. RESULTS AND CONCLUSION: (1) At 12 weeks after operation, three-dimensional reconstruction of Micro-CT showed that there was no obvious repair In the defect area of the model group, and there was still a large depression In the center. In the control group, the central depression area was significantly reduced and more trabecular structures of regenerated bone were observed. In the experimental group, the defect area was basically repaired. (2) At 12 weeks after operation, hematoxylin-eosin staining showed that there was no trabecular bone structure, disordered cell distribution and no cartilage lacunae at the defect area of the model group. In the control group, more bone tissue was reconstructed, and the defect area was mainly filled with cartilage-like tissue and fibrous tissue. In the experimental group, bone tissue was reconstructed adequately, and the defect area was mainly filled with chondroid cells and chondroid extracellular matrix. Cells arranged columnariy, similar to the surrounding cartilage. (3) At 12 weeks after surgery, Safranine O staining and collagen II immunohistochemical staining results showed that a small amount of glycosamlnoglycan was observed, but no type II collagen was found in the model group. The expression of glycosaminoglycan and type II collagen was more in the control group. The expression of glycosaminoglycan and type II collagen was highest In the experimental group compared with the other two groups. (4) The histological score of Wakitani soft tissue repair in the experimental group was higher than that in the control group and model group (P < 0.05). (5) The results suggested that Pluronic F-127 hydrogel complex loaded with SOX9 gene transfected bone marrow mesenchymal stem cells can promote the repair of cartilage defects.

Effect of parathyroid hormone on cartilage healing in a rabbit model of mandibular condylar fracture / 中国组织工程研究

BACKGROUND: A high fracture of the mandibular condyle is often accompanied by cartilage damage. At the same time, the muscle attached to the condyle is avulsed and becomes a free bone mass. How to speed up the concurrent healing of cartilage during bone healing has always been a clinical difficulty and challenge. OBJECTIVE: To investigate the effect of parathyroid hormone on the healing of condylar cartilage in rabbits with high condylar fracture after free reduction. METHODS: An experimental model of free reduction and fixation of condylar fracture was established in 48 New Zealand big-eared rabbits, which were randomly divided into experimental group and control group (n=20 per group). The experimental group was injected with parathyroid hormone (20 μg/kg) subcutaneously every other day, and the control group was injected with 1 mL of normal saline. The animals were sacrificed at 1, 2, 3, and 4 weeks postoperatively. The mandible condyle was histologically observed. Immunohistochemistry staining and PCR were used to detect the expression of Sox9 and matrix metalloproteinase 13 in the condylar cartilage. The study protocol was approved by the Animal Experimental Ethics Committee of Guizhou Medical University (approval No. 1700456). RESULTS AND CONCLUSION: The results of safranine O-fast green staining and hematoxylin-eosin staining indicated that there were more chondrocytes and cartilage matrix deposition in the experimental group than the control group. In the immunohistochemistry, the average absorbance of Sox9 in the experimental group was significantly higher than that in the control group within 1-3 postoperative weeks (P < 0.05). The average absorbance of matrix metalloproteinase 13 in the experimental group was lower than that in the control group within 1-3 postoperative weeks (P < 0.05). The expression of Sox9 mRNA in the experimental group was significantly higher than that in the control group within 1-3 postoperative weeks, P < 0.05). The expression of matrix metalloproteinase 13 mRNA in the experimental group was significantly lower than that in the control group within 1-3 postoperative weeks (P < 0.05). These findings indicate that intermittent subcutaneous injection of parathyroid hormone can up-regulate the expression of Sox9, inhibit the expression of matrix metalloproteinase 13, promote the transformation of mesenchymal stem cells to cartilage, and accelerate the repair of cartilage damage.

Correlation of Extracapsular Involvement of Lymph Node Metastasis With Prognosis and SOX9 Expression in Gastric Cancer Patients / 胃肠病学

Background: The extracapsular involvement (ECI) of lymph node metastasis has been considered as an important prognostic factor in patients with malignancies. Aims: To investigate the influence of ECI of lymph node metastasis on prognosis of patients with gastric cancer and its correlation with expression of transcriptional factor SOX9. Methods: A retrospective study was conducted in 200 consecutive gastric cancer patients with lymph node metastasis (TNM stage Ⅰ-Ⅲ) from Jan. 2010 to Dec. 2014 at Xi'an No. 1 Hospital. Patients were allocated into two groups according to the presence or absence of ECI, and their differences in clinicopathological characteristics, overall survival and SOX9 expression were compared. Univariate and multivariate analyses were performed by Cox proportional hazard model to identify the risk factors for poor prognosis. Results: ECI was associated with advanced T stage (T3, T4), N stage (N2, N3) and TNM stage (stage Ⅱ-Ⅲ), respectively (P<0.05). Furthermore, high expression of SOX9 was more frequently seen in metastatic lymph node with ECI than that without (94.5% vs. 58.3%, P<0.05). Kaplan-Meier survival analysis confirmed that patients in ECI group were associated with significantly shorter overall survival time (P<0.05). In multivariate analysis, TNM stage Ⅱ-Ⅲ (HR=3.224, 95% CI: 2.769-5.283, P<0.001), ECI of lymph node metastasis (HR=2.388, 95% CI: 1.802-3.209, P<0.001) and high expression of SOX9 in metastatic lymph node (HR=1.321, 95% CI: 1.201-1.684, P=0.001) were found to be the independent risk factors of poor overall survival in patients with gastric cancer. Conclusions: ECI of lymph node metastasis is associated with high expression of SOX9 in gastric cancer patients and can be used as an independent risk factor for poor prognosis. Also, ECI is a histomorphological indicator for invasive and metastatic phenotype of gastric cancer.

Determination of SOX9 Gene Expression In Brain Tumours Among East Coast Malaysia Population

@#Introduction: SOX9, a members of SOX family, plays a significant roles in developmental processes during embryogenesis, including brain tissue. Few studies have shown that SOX9 has been involved in tumourigenesis of several types of cancer including brain tumour. However, such studies are still lacking in the Malaysian population. The aim of this study was to determine SOX9 expression level in several types of brain tumours in East Coast Malaysia. Materials and Methods: Five formalin-fixed pariffin-embedded brain tumour samples of Malay descendants were sectioned by using microtome. RNA extraction was performed with slight modification by adding Trizol during tissue lysis. The RNA was converted to cDNA using reverse transcription technique before SOX9 expression was detected using RT q-PCR assay in brain tumours normalized to non-neoplastic brain tissues. Results: Overall results displayed that SOX9 gene in all samples were up-regulated. SOX9 overexpression was found in both high and low grade glioma (anaplastic and pilocytic astrocytoma respectively). This is consistence with both low grade (benign) and atypical meningioma. Secondary brain tumour also showed up-regulation when compared to normal brain tissue. Conclusion: Up-regulation in SOX9 expression in selected brain tumours in Malay patients revealed its significant roles in brain tumourigenesis. Functional studies should be carried out to observe the SOX9 functions and mechanism whether they should reflect their diverse roles in Malaysia population

SOX9 promotes epithelial-mesenchymal transition in non-small cell lung cancer A549 cells via Wnt/β-catenin pathway / 中国肿瘤生物治疗杂志

@#Objective:To explore the mechanism by which SRY-related high mobility group-box 9 (SOX9) promotes the epithelial mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC) A549 cells via the Wnt/β-catenin pathway. Methods: The human NSCLCA549 cell line was divided into three groups: OE-NC group, OE-SOX9 group and OE-SOX9+XAV-939 group. The cells in OESOX9 group were transfected with SOX9 pcDNA plasmid to up-regulate the expression level of SOX9; The cells in OE-SOX9+XAV939 group were transfected with SOX9 pcDNA plasmid while the β-catenin inhibitor XAV-939 (1.0 μmol/L) was added to the medium. qPCR was used to detect SOX9 mRNA levels; CCK-8 was used to examine the proliferation of A549 cells; Wound-healing assay and Transwell chamber assay were used to detect the migration and invasion ofA549 cells, respectively; and WB was used to detect protein expressions of SOX9, β-catenin, E-cadherin, γ-catenin, N-cadherin and vimentin. Results: The mRNA and protein levels of SOX9 in OE-SOX9 group and OE-SOX9+XAV-939 group were significantly higher than those in the OE-NC group after transfection (all P< 0.05), while there was no significant difference between the OE-SOX9 group and the OE-SOX9+XAV-939 group (P>0.05). The proliferation, migration and invasion of cells in OE-SOX9 group were significantly higher than those in OE-NC group; however, those abilities in OE-SOX9+XAV-939 group were significantly lower than those in OE-SOX9 group (all P<0.05). The level of β-catenin protein in OE-SOX9 group was significantly higher than that in the OE-NC group, while the level of β-catenin protein in OE-SOX9+XAV-939 group was lower than that in OE-SOX9 group (all P<0.05). Compared with the OE-NC group, the levels of phenotypic markers of epithelial cells, E-cadherin and γ-catenin, were down-regulated, and the phenotypic markers of mesenchymal cells, N-cadherin and vimentin, were up-regulated in cells of OE-SOX9 group; however, E-cadherin and γ-catenin were higher, and N-cadherin and vimentin were lower in OE-SOX9+XAV-939 group than those in OE-SOX9 group (all P<0.05). Conclusion: SOX9 could promote proliferation, migration and EMT of NSCLCA549 cells by activating the Wnt/β-catenin pathway. ··

The research progress of Sox9 gene on pancreatic development / 国际儿科学杂志

Sox9 is a member of the Sox family of E groups,and it plays a role in gender determination and differentiation,tumor development and organ development.In the process of pancreatic development,Sox9 and its crosstalk with the Notch,Wingless and INT-1 (Wnt) and fibroblast growth factor (Fgf) pathways are involved in the proliferation and differentiation of early pancreatic endocrine progenitor cells.It is a versatile factor of Sox9 in the regulation network of pancreatic endocrine development.Endocrine beta cells are the only cells that secrete insulin.The absolute or relative inadequate of insulin will lead to increase blood glucose levels and then lead to the occurrence of diabetes.The study of pancreatic endocrine development provides a new idea to explore the pathogenesis of diabetes.In this paper,the effects of Sox9 and its transcriptional regulation mechanisms on pancreatic endocrine development are summarized.

Construction of Lentiviral Expression Vector Carrying SOX9 Gene and Its Stable Expression in LNcap Cell / 昆明医科大学学报

Objective To construct the lentiviral expression vector of SOX9 gene and establish a LNcap cell strain with stable expression of SOX9 . Methods SOX9 gene was amplified by PCR and cloned into lentiviral expression vector pLVX-IRES-Puro. pLVX-IRES-FLAG-SOX9 recombinant plasmid was verified by restriction enzyme digestion and DNA sequencing. Then we gained recombinant virus particles in packaging cell HEK 293 and infected LNcap cell. The monoclonal LNcap cell strain stably expressed were obtained through puromycin screening. The mRNA level and protein level in the infected LNcap cell were detected by qRT-PCR and Western blot respectively. Results Restriction enzyme digestion and sequencing demonstrated that SOX9 cDNA was successfully cloned into pLVX-IRES-FLAG lentiviral vector. After the transfection to LNCaP cells, the monoclonal cell strain of stably expressed SOX9 were obtained by puromycin screening, which showed expression of SOX9 mRNA detected by qRT-PCR. Western blot analysis revealed that SOX9 protein expressed markedly. Conclusion The recombinant lentiviral vector bearing human SOX9 cDNA has been successfully constructed, and the exogenous expression of SOX9 in LNcap cells was achieved.

Effect of mechanical environment on differentiation of mesenchymal stem cells at fracture site during early stage of fracture healing and research of related mechanisms / 中国药理学通报

Aim To explore the influence of mechani-cal environment on differentiation of mesenchymal stem cells ( MSCs) at fracture site during early stage of frac-ture healing. Methods The rat femur fracture models were established and fixed with external fixation frame of different stiffness coefficient and were divided into three groups, namely, the sham group with super-high stiffness external fixation frame, the high stiffness fixa-tion frame group and the low stiffness fixation frame group. The bone callus and the degree of fracture heal-ing after two weeks compared with the very day after surgery were observed by X-ray. And the effects were evaluated respectively by histopathology 6 weeks after operation. Callus tissues of 2 days, 6 days, 10 days, 14 days after operation were acquired. Runx2, Osterix and Sox9 expression were measured by RT-qPCR. Runx2 and Sox9 were tested by Western blot. Results X-ray showed that the amount of callus of high stiff-ness group was significantly less than that of low stiff-ness group. Histological study showed, sham group and high stiffness group achieved less callus and had lots of active bone cells while low stiffness group got cartilage ossification well. The results of RT-qPCR showed that the level of Runx2 , Osterix increased with-in 14 days. And Sox9 reached the highest level in sham group and high stiffness group 10 days after sur-gery and declined in 14 days while low stiffness group had been rising within 14 days after surgery. The ex-pression levels of Runx2 , OSX and Sox9 in 14 days af-ter operation presented statistically significant differ-ence among the three groups at the same time point, i. e. low stiffness group > high stiffness group > the sham group ( P <0.05 ) . The results of Western blot were consistent with those of RT-qPCR. Conclusion The differentiation of MSCs at fracture site is affected by the mechanical environment, and subtle movement could promote the healing of fracture sites.

Relationship between expression level of sex-determining region Y box 9 and metastasis of oral squamous cell carcinoma / 中华口腔医学杂志

Objective@#To investigate the effects of sex-detemining region Y box9 (SOX9) expression levels on the proliferation, migration and metastasis in oral squamous cell carcinoma (OSCC).@*Methods@#A total of 74 OSCC pathological specimens were collected from Shanghai OSCC Tissue and Biological Informations Bank, and clinicopathological information of these specimens were collected. Immunohistochemistry assay was used to examine the expression levels of SOX9 in OSCC and to analyze their relationship with clinicopathological features. Cell counting kit-8 assay and cloning formation was used to observe the relationship between the expression levels of SOX9 and the proliferation of OSCC. Transwell experiment and scratch test were used to detect the difference of the ability of OSCC in cell lines with different expression levels of SOX9.@*Results@#The risk of lymph node metastasis in patients with high expression of SOX9 was significantly increased (P=0.010). In the Transwell experiment, the number of HN6 cells (671.0±57.4, P=0.000) migrated to the lower chamber more than that of CAL27 cells (172.0±13.9). In the scratch experiment, HN6 cells [0 h: (93.7±2.1) μm; 6 h: (56.7±2.5) μm; 12 h: (29.7±3.1) μm] migrated faster than CAL27 [0 h: (93.7±1.5) μm; 6 h: (78.0±2.0) μm; 12 h: (42.0±3.0) μm](P<0.05). The migration ability of the cell line (HN6) with high-expression of SOX9 was significantly higher than that in cell line (CAL27) with low-expression SOX9 (P<0.05). The expression levels of SOX9 in OSCC were no significant on cell proliferation (P>0.05).@*Conclusions@#High expression of SOX9 can promote the migration and lymph node metastasis of OSCC. SOX9 is a candidate gene target for the diagnosis and intervention of lymph node metastasis in OSCC.

Transcriptional Network Controlling Endochondral Ossification

Endochondral ossification is the fundamental process of skeletal development in vertebrates. Chondrocytes undergo sequential steps of differentiation, including mesenchymal condensation, proliferation, hypertrophy, and mineralization. These steps, which are required for the morphological and functional changes in differentiating chondrocytes, are strictly regulated by a complex transcriptional network. Biochemical and mice genetic studies identified chondrogenic transcription factors critical for endochondral ossification. The transcription factor sex-determining region Y (SRY)-box 9 (Sox9) is essential for early chondrogenesis, and impaired Sox9 function causes severe chondrodysplasia in humans and mice. In addition, recent genome-wide chromatin immunoprecipitation-sequencing studies revealed the precise regulatory mechanism of Sox9 during early chondrogenesis. Runt-related transcription factor 2 promotes chondrocyte hypertrophy and terminal differentiation. Interestingly, endoplasmic reticulum (ER) stress-related transcription factors have recently emerged as novel regulators of chondrocyte differentiation. Here we review the transcriptional mechanisms that regulate endochondral ossification, with a focus on Sox9.

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.

The effects of dynamic pressure on expression of Sox9 mRNA and protein in metaphyseal chondrocytes of rats / 天津医药

Objective To study the effect of dynamic stress stimulation on the expression of Sox9 mRNA and protein in metaphyseal chondrocytes in vitro, and to explore the specific mechanism of mechanical signal transduction. Methods The rat metaphyseal chondrocytes separated and cultured for the 3rd generation in vitro were randomly divided into four groups:control group (all interventions were not applied), simple dynamic pressure group (a dynamic pressure stimulus with a size of 90 mmHg and a frequency of 0.1 Hz was applied using an open pressure control culture system), simple calcium antagonist group (the concentration of 10μmol/L nifedipine was given) and dynamic pressure+calcium antagonist group (a dynamic pressure stimulus with a size of 90 mmHg, frequency of 0.1 Hz and concentration of 10 μmol/L nifedipine were given at the same time). The expression of Sox9 mRNA was detected after 24 h intervention by real-time quantitative polymerase chain reaction (RT-PCR) in four groups. The expression of Sox9 protein was detected by Western blot assay. The intracellular free Ca2+ in metaphyseal chondrocytes was labeled with Fluo-3/AM, and the average fluorescence intensity detected by laser scanning confocal scanning microscopy was compared between four groups. Results The expression of Sox9 mRNA was 3.81 times higher in dynamic stress group than that in the control group, and the protein expression level was 2.33 times higher than that of the control group (P<0.05). There were no significant differences in the expression of Sox9 mRNA and protein between the calcium antagonist group and the control group. The expressions of Sox9 mRNA and protein were lower in dynamic pressure+calcium antagonist group than those in the dynamic stress group, but which were higher than those of control group(P<0.05). The results of average fluorescence intensity showed that there was no significant difference in the intracellular free Ca2+ concentration between four groups (P > 0.05). Conclusion Dynamic stress stimulation can increase the expression of Sox9 mRNA and protein in rat metaphyseal chondrocytes. There is calcium channel involvement in the mechanical signal transduction.

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.