Objective To investigate the expression of stromal cell-derived factor-1 (SDF-1) and its clinical significance in blood plasma of patients with breast tumor. Methods The level of SDF-1 protein was examined by enzyme linked immunosorbent assay (ELISA) in blood plasma of 26 patients with breast benign tumor and 52 patients with breast cancer. Results The SDF-1 protein in blood plasma was detected in both breast benign tumor patients and breast cancer ones. The level of SDF-1 protein in patients with breast cancer was higher than that in ones with breast benign tumor, and there was a statistical difference between them （P=0.000）. In patients with breast cancer, the level of SDF-1 protein in axillary lymph node (ALN) metastasis positive patients was significantly higher than that in ALN metastasis negative ones （P=0.036）. Conclusion The level of SDF-1 protein in blood plasma may be a specific tumor marker. Its level is correlated with lymph node involvement in breast cancer.
Objective To investigate the influence on matrix metalloproteinases (MMP) 3, 9, and 13 levels of human articular cartilage cells after blocking stromal cell derived factor 1 (SDF-1)/ chemokine receptor 4 (CXCR4) signaling pathway withAMD3100 and to define the function mechanism of AMD3100. Methods A total of 144 cartilage blocks from 12 osteoarthritis (OA) patients undergoing total knee arthroplasty (OA cartilage group) and 144 normal cartilage blocks (Mankin score of 0 or 1) from 12 patients undergoing traumatic amputation (normal cartilage group). OA cartilage group was further divided into subgroups A1, B1, and C1, and normal cartilage group into subgroups A2, B2, and C2. The cartilage tissues were cultured in DMEM solution containing 100 ng/mL SDF-1 and 1 000 nmol/L AMD3100 in subgroup A, 100 ng/mL SDF-1 and 1 000 nmol/L MAB310 in subgroup B, and 100 ng/mL SDF-1 in subgroup C, respectively. The levels of MMP-3, 9, and 13 were measured by ELISA; the expressions of MMP-3, 9, and 13mRNA were tested by RT-PCR. Results ELISA and RT-PCR results showed that the levels of MMP-3, 9, and 13 and the expressions of MMP-3, 9, and 13 mRNA were significantly lower in subgroup A than in subgroups B and C at the same time points (P lt; 0.05); the levels of MMP-3, 9, and 13 and the expressions of MMP-3, 9, and 13 mRNA were significantly higher in OA cartilage group than in normal cartilage group at the same time points (P lt; 0.05). Conclusion SDF-1 could induce overexpression and release of MMP-3, 9, and 13 in the articular cartilage through the SDF-1/CXCR4 signaling pathway; AMD3100 could reduce the mRNA expressions and secretion of MMP-3, 9, and 13 in OA cartilage by blocking the SDF-1/CXCR4 signaling pathway; but AMD3100 could not make the secretion of MMP-3, 9, and 13 return to normal levels in OA cartilage.
Objective To review research progress of adipose tissuederived stromal cells (ADSCs).Methods The recent articles on ADSCs were extensively reviewed, and the culture and differentiation ability of ADSCs were investigated.Results A population of stem cells could be isolated from adult adipose tissue, they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling. The majority of the isolated cells were mesenchymal origin, with a few pericytes,endothelial cells and smooth muscle cells. ADSCs could be induced to differentiate intomultiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic and adipogenic cells, they could also differentiate into nerve cells.Conclusion ADSCs can substitute mesenchymal stem cells and become an alternative stem cells source for tissue engineering.
The osteogenc potential of bone marrow has been proved by experiment. To investigate more in details, bone marrow was obtained from the trochanteric region of femur of NewZealand rabbit in 4 to 8 weeks old. After being cultured in vitro for one week, the hematopoietic component of the bone marrow had disappeared, thus the stromal cells were obtained. Then the stromal cells were subcultured in cultural fluid containing dexamethasone (10-8 mol/L) and natrium glycerophosphate (10mmol/L). Under the phasecontrast microscope, it was found that being cultured for 15 days. The stromal cells were lined up in one layer and late the secretion activity was increased and gradually transformed into multilayer structure and was congregated into diffused opaque clusters in twenty days. During culture, the cells were examined by tetracycline fluorescence label, histochemistry stains, transmission electron microscopy, scanning electron microscopy and energy dispersive X-ray microanalysis. The results showed that the morphological and biological characteristics of the cultured stromal cells derived from the bone marrow were similiar to those of osteoblasts and could synthesized mineralized new bone tissue in vitro.
【Abstract】Objective Stromal cell-derived factor-1(SDF-1, CXCL12) is a member of the CXC subfamily of chemokines which, through its cognate receptor (CXCR4), plays an important role in tumor invasion and metastasis. This study analyzed quantitatively the expression of SDF-1 and its relation with clinicopathologic feature and clinical outcome in human breast cancer.Methods Expression of SDF-1 mRNA in 8 breast cancer cell lines, an endothelial cell line HECV and a fibroblast cell MRC5 was studied by using RT-PCR. In addition, the expression of SDF-1 was investigated at both protein (immunohistochemistry) and mRNA(real-time PCR) levels in a group of human normal mammary(n=32) and tumour tissues(n=120). Results SDF-1 expression was identified in MRC5, MDA-MB435s, MDA-MB436, MCF7 cell lines, breast tumour and normal tissues. Significantly higher level of SDF-1 was seen in lymph node positive than in lymph node negative tumours (399.00±210.00 vs 0.89±0.47), P=0.048. The level of SDF-1 expression in patients who developed local recurrence or metastasis, or patients who died of breast cancer was higher than in patients who were disease free as well, (670.00±346.00 vs 0.83±0.35), P=0.01. It was most notable that level of SDF-1 was significantly correlated with over survival (P=0.01) and incidence free survival (P=0.035, by Cox proportion analysis).Conclusion SDF-1 is a factor that is expressed in both stromal cells and some breast cancer cells. Its level are correlated with lymph node involvement, prognosis and survival in patients with breast cancer. SDF-1 may therefore have a potential prognostic value in breast cancer.
Objective To investigate the expression of stromal cell derived factor-1 ( SDF-1) and the effects of budesonide suspension for inhalation ( Pulmicort Respules) in mice with asthma. Methods Thirty Kunming female mice were randomly divided into three groups, ie. a control group, an asthma group, and a pulmicort treatment group. The asthma group and the pulmicort treatment group were sensitized with ovalbumin ( OVA) by a combination of intraperitoneal injection and repeated OVA intranasal challenges to establish mouse asthma model. The pulmicort treatment group received 100μL pulmicort by intranasal administration before OVA challenge. The immunohistochemistry was used to estimate the expression of SDF-1 in lung tissues. HE staining and Wright-Giemsa staining method were used to assess inflammatory infiltration in the airway and bronchoalveolar lavage fluid ( BALF) respectively. Results The expression of SDF-1 in the asthma group increased significantly compared with the control group ( 0.48 ±0.03 vs. 0.21 ± 0.02, Plt;0.05) , and significantly decreased after the intervention with pulmicort ( 0.29 ±0.01 vs. 0.48 ± 0.03, Plt; 0.05 ) . Compared with control group, the infiltration of inflammatory cells in airway was significantly enhanced in the asthma group, and attenuated in the pulmicort treatment group. The total number of inflammatory cells and eosinophil, lymphocyte, neutrophil counts in BALF increased significantly in the asthma group compared with the control group, and decreased significantly after pulmicort intervention. Conclusion SDF-1 may play an important role in the recruitment of inflammatory cells in asthmatic airway and pulmicort may relieve airway inflammation by decreasing the expression of SDF-1.
ObjectiveTo explore effect and mechanism of the carcinoma associated fibroblasts (CAFs) of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast cancer cells. MethodsBreast cancer cell line of MDA-MB-231 (abbreviated as MDA), CAFs, and normal breast tissue fibroblasts (NFs) of the same breast cancer patient were collected, and mixed with normal saline (NS) or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka (AMD3100, abbreviated as AMD) for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups:MDA+NS group, NFs+NS group, MDA+NFs+NS group, MDA+NFs+AMD group, MDA+CAFs+AMD group, and MDA+CAFs+NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein in tumor specimens were detected by using real-time PCR and Western blot method respectively. ResultsExcept for NFs+ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA+CAFs+NS group, the volume of tumor[(9.092±2.662) cm3], level of plasma SDF-1[(75.25±16.23) ng/L], and expression levels of SDF-1 mRNA (the median level was 14.714) and protein (the median level was 0.673). of tumor tissue were significantly greater or higher than those of the other 5 groups (P < 0.050). In addition, lymph node metastasis were found in 4 mice in MDA+CAFs+NS group, and 2 in MDA+NS group. The tumor metastasis of lung and liver was not found in all nude mice. ConclusionsCAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 (CXCR4), signal pathway.
ObjectiveTo detect the content of stromal cell derived factor-1α(SDF-1α) in peripheral blood of patients with gastric adenocarcinoma (GC) and investigate its clinical significances. MethodsThe contents of SDF-1αin the peripheral blood of 90 patients with GC were detected by enzyme-linked immunosorbent assay. The correlation of SDF-1αcontent with the clinicopathologic parameters and prognosis after operation were analyzed. Results①The content of SDF-1αin the patients with GC[(6950.8±1131.3) ng/L] was significantly higher than that in the normal healthy volunteers[(5023.7±1103.8) ng/l, P=0.036].②The content of SDF-1αin the GC patients with distant metastasis[(8251.6±1042.5) ng/L] was significantly higher than that without distant metastasis[(6785.3±1025.0) ng/L, P < 0.001]. The contents of SDF-1αin the peripheral blood of patients with distant metastasis either in the liver (P=0.002) or in the lung (P=0.030) were significantly higher than those without distant metastasis (liver or lung).③The TNM stage was later (P < 0.001), lymph node metastasis was broader (P=0.018), invasion of tumor was deeper (P < 0.001), vascular invasion (P < 0.001) and lymphatic vessel invasion were present (P < 0.001), the contents of SDF-1αwere higer. Logistic regression analysis revealed that the depth of tumor invasion (OR=14.999, 95% CI 3.568-74.456, P=0.027) and distant metastasis (OR=0.186, 95% CI 0.610-2.014, P=0.026) were correlated with the high SDF-1αcontent.④The survival time of the patients with higher content of SDF-1αwas significantly shorter than that of the lower content of SDF-1α(P < 0.001). Cox proportial hazard regression model analysis demonstrated that TNM stage (RR=2.497, 95% CI 1.987-10.238, P=0.009), vascular invasion (RR=7.501, 95% CI 2.086-16.942, P=0.002), and high content of SDF-1α(RR=18.302, 95% CI 6.895-30.538, P=0.001) in the peripheral blood were the independent risk factors for survival of the patients with GC. ConclusionHigh content of SDF-1αin peripheral blood might suggest the occurrence of lymph node metastasis, hepatic metastasis or lung metastasis and indicate the poorer prognosis of GC.
ObjectiveTo observe the change of stromal cell-derived factor 1α/cysteine X cysteine receptor 4 (SDF-1α/CXCR4) signaling pathway during the process of axial stress stimulation promoting bone regeneration, and to further explore its mechanism.MethodsA total of 72 male New Zealand white rabbits were selected to prepare the single cortical bone defect in diameter of 8 mm at the proximal end of the right tibia that repaired with deproteinized cancellous bone. All models were randomly divided into 3 groups (n=24). Group A was treated with intraperitoneally injection of PBS; Group B was treated with stress stimulation and intraperitoneally injection of PBS; Group C was treated with stress stimulation and intraperitoneally injection of AMD3100 solution. The X-ray films were taken and Lane-Sandhu scores of bone healing were scored at 2, 4, 8, and 12 weeks after operation, while specimens were harvested for HE staining, immunohistochemical staining of vascular endothelial growth factor (VEGF) and CXCR4, and Western blot (SDF-1α and CXCR4). The bone healing area was scanned by Micro-CT at 12 weeks after operation, and the volume and density of new bone were calculated.ResultsX-ray film showed that the Lane-Sandhu scores of bone healing in group B were significantly higher than those in groups A and C at 4, 8, and 12 weeks after operation (P<0.05). Micro-CT scan showed that the bone defect was repaired in group B and the pulp cavity was re-passed at 12 weeks after operation. The volume and density of new bone were higher in group B than in groups A and C (P<0.05). HE staining showed that the new bone growth in bone defect area and the degradation of scaffolds were faster in group B than in groups A and C after 4 weeks. The immunohistochemical staining showed that the expressions of VEGF and CXCR4 in 3 groups reached the peak at 4 weeks, and group B was higher than groups A and C (P<0.05). Western blot analysis showed that the expressions of SDF-1α and CXCR4 in group B were significantly higher than those in groups A and C at 4 and 8 weeks after operation (P<0.05).ConclusionAxial stress stimulation can promote the expression of SDF-1α in bone defect tissue, activate and regulate the CXCR4 signal collected by marrow mesenchymal stem cells, and accelerate bone regeneration in bone defect area.
ObjectiveTo explored the effect of stromal cell-derived factor 1α (SDF-1α) on promoting the migration ability of rat adipose derived stem cells (rADSCs) by constructed the rADSCs overexpression SDF-1α via adenovirus transfection.MethodsrADSCs were isolated from adipose tissue of 6-week-old SPF Sprague Dawley rats. Morphological observation, multi-directional differentiations (osteogenic, adipogenic, and chondrogenic inductions), and flow cytometry identification were performed. Transwell cell migration experiment was used to observe and screen the optimal concentration of exogenous SDF-1α to optimize the migration ability of rADSCs; the optimal multiplicity of infection (MOI) of rADSCs was screened by observing the cell status and fluorescence expression after transfection. Then the third generation of rADSCs were divided into 4 groups: group A was pure rADSCs; group B was rADSCs co-cultured with SDF-1α at the best concentration; group C was rADSCs infected with recombinant adenovirus-mediated green fluorescent protein (Adv-GFP) with the best MOI; group D was rADSCs infected with Adv-GFP-SDF-1α overexpression adenovirus with the best MOI. Cell counting kit 8 (CCK-8) and Transwell cell migration experiment were preformed to detect and compare the effect of exogenous SDF-1α and SDF-1α overexpression on the proliferation and migration ability of rADSCs.ResultsThe cell morphology, multi-directional differentiations, and flow cytometry identification showed that the cultured cells were rADSCs. After screening, the optimal stimulating concentration of exogenous SDF-1α was 12.5 nmol/L; the optimal MOI of Adv-GFP adenovirus was 200; the optimal MOI of Adv-GFP-SDF-1α overexpression adenovirus was 400. CCK-8 method and Transwell cell migration experiment showed that compared with groups A and C, groups B and D could significantly improve the proliferation and migration of rADSCs (P<0.05); the effect of group D on enhancing the migration of rADSCs was weaker than that of group B, but the effect of promoting the proliferation of rADSCs was stronger than that of group D (P<0.05).ConclusionSDF-1α overexpression modification on rADSCs can significantly promote the proliferation and migration ability, which may be a potential method to optimize the application of ADSCs in tissue regeneration and wound repair.