Stromal Cell-derived Factor-1 mRNA Expression Research Articles

Prognostic role of the stromal cell derived factor-1 in patients with hepatitis B virus-related acute-on-chronic liver failure.

Stromal cell derived factor-1 (SDF-1) plays a pivotal role in the recruitment of stem cells to injured livers. However, the changes of SDF-l in patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) have yet to be elucidated. To study the SDF-1 changes in patients with HBV-related ACLF. 30 patients with HBV-related ACLF, 27 patients with chronic hepatitis B and 20 healthy individuals are involved in our study. The SDF-l mRNA expression in liver tissue was detected by quantitative real-time polymerase chain reaction. Immunohistochemical staining was performed to illustrate the expression of SDF-l, CXC receptor 4 (CXCR4) and Ki67. The serum SDF-l concentrations were also detected by enzyme-linked immunosorbent assays. The expression of SDF-1 mRNA from ACLF patients was remarkably higher than that from other patients (both P < 0.05). The expression of SDF-l, CXCR4 and Ki67 from ACLF were the highest among the three groups (all P < 0.01). The serum SDF-l levels in ACLF patients were significantly lower than that in other patients (both P < 0.01). Moreover, in ACLF patients, the serum SDF-1 Levels were positively correlated with serum total bilirubin and international normalized ratio. In addition, the serum SDF-l levels in survival were significantly lower compared with the non-survivals (P < 0.05). The area under the curve for the serum SDF-1 level in predicting 28-d mortality was 0.722 (P < 0.05). This study provides the SDF-1 changes in patients with HBV-related ACLF. The SDF-1 Level at admission may serve as a promising prognostic marker for predicting short-term prognosis.

Effects of sitagliptin activation of the stromal cell-derived factor-1/CXC chemokine receptor 4 signaling pathway on the proliferation, apoptosis, inflammation, and osteogenic differentiation of human periodontal ligament stem cells induced by lipopolysaccharide.

This study aimed to investigate the effects of sitagliptin on the proliferation, apoptosis, inflammation, and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in lipopolysaccharide (LPS)-induced inflammatory microenvironment and its molecular mechanism. hPDLSCs were cultured in vitro and treated with different concentrations of sitagliptin to detect cell viability and subsequently determine the experimental concentration of sitagliptin. An hPDLSCs inflammation model was established after 24 h of stimulation with 1 µg/mL LPS and divided into blank, control, low-concentration sitagliptin (0.5 µmol/L), medium-concentration sitagliptin (1 µmol/L), and high-concentration sitagliptin (2 µmol/L), high-concentrationsitagliptin+stromal cell derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) pathway inhibitor (AMD3100) (2 µmol/L+10 µg/mL) groups. A cell-counting kit-8 was used to detect the proliferation activity of hPDLSCs after 24, 48, and 72 h culture. The apoptosis of hPDLSCs cultured for 72 h was detected by flow cytometry. After inducing osteogenic differentiation for 21 days, alizarin red staining was used to detect the osteogenic differentiation ability of hPDLSCs. The alkaline phosphatase (ALP) activity in hPDLSCs was determined using a kit. The levels of inflammatory factors [tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6] in the supernatant of hPDLSCs culture were detected by enzyme-linked immunosorbent assay. The mRNA expressions of osteogenic differentiation genes [Runt-associated transcription factor 2 (RUNX2), osteocalcin (OCN), osteopontin (OPN)], SDF-1 and CXCR4 in hPDLSCs were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Western blot analysis was used to determine SDF-1 and CXCR4 protein expression in hPDLSCs. Compared with the blank group, the proliferative activity, number of mineralized nodules, staining intensity, ALP activity, and RUNX2, OCN, OPN mRNA, SDF-1, and CXCR4 mRNA and protein expression levels of hPDLSCs in the control group significantly decreased. The apoptosis rate and levels of TNF-α, IL-1β, and IL-6 significantly increased (P<0.05). Compared with the control group, the proliferative activity, number of mineralized nodule, staining intensity, ALP activity, and RUNX2, OCN, OPN mRNA, SDF-1, and CXCR4 mRNA and protein expression levels of hPDLSCs in low-, medium-, and high-concentration sitagliptin groups increased. The apoptosis rate and levels of TNF-α, IL-1β, and IL-6 decreased (P<0.05). AMD3100 partially reversed the effect of high-concentration sitagliptin on LPS-induced hPDLSCs (P<0.05). Sitagliptin may promote the proliferation and osteogenic differentiation of hPDLSCs in LPS-induced inflammatory microenvironment by activating the SDF-1/CXCR4 signaling pathway. Furthermore, it inhibited the apoptosis and inflammatory response of hPDLSCs.

Cholesterol modification of SDF-1-specific siRNA enables therapeutic targeting of angiogenesis through Akt pathway inhibition

Neovascularization during ocular tissue repair can cause severe visual loss in the optical axis and is therefore an issue of considerable concern to ophthalmologists. Here, we introduced a cholesterol-modified siRNA delivery system targeting stromal cell-derived factor 1 (SDF-1) to treat ocular angiogenesis in vivo. SDF-1 expression was analyzed in rat endothelial progenitor cells (EPCs) and bone marrow mesenchymal stem cells (BMSCs) using quantitative PCR (qPCR). Migration ability of BMSC and HUVEC were assessed through transwell assay. The proliferation effect of chol-siSDF1 on HUVEC was measured by colony formation assay. In vivo anti-angiogenic effects of chol-siSDF1 were tested in a cornea alkali burn model and the area of cornea neovascularization was measured using computer-imaging analysis system. Then phosphorylated Akt and total Akt protein levels were measured through western blot. Results turned out that rat EPCs and BMSCs showed high SDF-1 mRNA expression, which can be down-regulated by using chol-siSDF-1. Chol-siSDF-1 could significantly inhibit migration of BMSC and HUVEC. In addition, chol-siSDF1 also could inhibit HUVEC proliferation and exert a significant anti-angiogenic effect in corneal alkali burn model. As for the mechanism, chol-siSDF1 may inhibit the neovascularization, proliferation and metastasis through inhibiting the Akt signaling pathway. Thus, cholesterol modification of siRNA targeting SDF-1 displays an effective inhibition of migration and angiogenesis, with a much longer duration of inhibition effect.

Role of SDF-1:CXCR4 in Impaired Post-Myocardial Infarction Cardiac Repair in Diabetes.

Diabetes is a risk factor for worse outcomes following acute myocardial infarction (AMI). In this study, we tested the hypothesis that SDF‐1:CXCR4 expression is compromised in post‐AMI in diabetes, and that reversal of this defect can reverse the adverse effects of diabetes. Mesenchymal stem cells (MSC) isolated from green fluorescent protein (GFP) transgenic mice (control MSC) were induced to overexpress stromal cell‐derived factor‐1 (SDF‐1). SDF‐1 expression in control MSC and SDF‐1‐overexpressing MSC (SDF‐1:MSC) were quantified using enzyme‐linked immunosorbent assay (ELISA). AMI was induced on db/db and control mice. Mice were randomly selected to receive infusion of control MSC, SDF‐1:MSC, or saline into the border zone after AMI. Serial echocardiography was used to assess cardiac function. SDF‐1 and CXCR4 mRNA expression in the infarct zone of db/db mice and control mice were quantified. Compared to control mice, SDF‐1 levels were decreased 82%, 91%, and 45% at baseline, 1 day and 3 days post‐AMI in db/db mice, respectively. CXCR4 levels are increased 233% at baseline and 54% 5 days post‐AMI in db/db mice. Administration of control MSC led to a significant improvement in ejection fraction (EF) in control mice but not in db/db mice 21 days after AMI. In contrast, administration of SDF‐1:MSC produced a significant improvement in EF in both control mice and db/db mice 21 days after AMI. The SDF‐1:CXCR4 axis is compromised in diabetes, which appears to augment the deleterious consequences of AMI. Over‐express of SDF‐1 expression in diabetes rescues cardiac function post AMI. Our results suggest that modulation of SDF‐1 may improve post‐AMI cardiac repair in diabetes. stem cells translational medicine 2018;7:115–124

Neuroprotection by intravenous transplantation of bone marrow mononuclear cells from 5-fluorouracil pre-treated rats in a model of ischemic stroke

Our previous studies showed that bone marrow mononuclear cells (BMMNCs) from 5-fluorouracil (5-FU) pre-treated rats (named BMRMNCs) had a better therapeutic efficacy in ischemia/reperfusion rats as compared to BMMNCs from untreated rats. This study was undertaken to further explore the potential mechanisms underlying the neuroprotective effects of BMRMNCs in the same model. Rats were intravenously pre-treated with 5-FU, and BMRMNCs were collected 7 days later and subjected to flow cytometry for detection of CD34, CD45 and CD90. Middle cerebral artery occlusion (MCAO) was induced in rats, and BMMNCs and BMRMNCs were independently transplanted via the tail vein at 24 h after MCAO. NISSL staining was performed 14 days after cell transplantation and the viable cells in the hippocampus were counted. Stromal cell-derived factor 1 (SDF-1) mRNA expression was detected in the penumbra at 7 and 14 days after treatment. The contents of pro-inflammatory cytokines and growth factors as well as microvessel density (MVD) were determined at 14 days. Results showed more BMRMNCs were positive for CD34, CD45 and CD90. After transplantation, more viable cells were observed in the hippocampus of BMRMNCs treated rats. In addition, BMRMNCs transplantation significantly increased MVD, reduced pro-inflammatory cytokines and raised growth factors in the penumbra. However, the SDF-1 mRNA expression was comparable between BMRMNCs group and BMMNCs group. Our results indicate that BMRMNCs are likely to more effectively improve the local microenvironment to increase viable cells and elevate angiogenesis, exerting neuroprotective effects on cerebral ischemia in rats.

Effect of lipopolysaccharide on the stromal cell-derived factor-1 expression in human stem cells from apical papilla

To investigate the expression of stromal cell-derived factor-1 (SDF-1) in human stem cells from apical papilla (SCAP), and to evaluate the effect of lipopolysaccharide (LPS) on SDF-1 expression by SCAP. SCAP were isolated from dental papilla of human immature third molars. The expression of SDF-1 was evaluated by reverse transcription-PCR (RT-PCR). After SCAP being exposed to different concentrations (0.1, 1.0, 10 mg/L) of LPS for 24 and 48 h, the effect of LPS on cell proliferation and gene expression of SDF-1 was investigated by cell counting kit-8 and real-time PCR respectively, while cells without LPS stimulation were considered as negative control. LPS had no significant effect on SCAP proliferation until day 7. RT-PCR assays demonstrated that SCAP expressed SDF-1 mRNA. Different concentrations of LPS significantly promoted the SDF-1 expression in SCAP after 24 h (F = 12.102, P = 0.002) and 48 h (F = 39.054, P < 0.001) exposure, with relative gene expression ratio (experimental/control) increased to 1.4 ± 0.1, 2.2 ± 0.4, 2.3 ± 0.5 in 24 h group and 2.1 ± 0.4, 3.4 ± 0.3, 3.8 ± 0.5 in 48 h group. Isolated SCAP in cultures have the expression of SDF-1 mRNA. LPS can significantly promote the expression of SDF-1 in SCAP.

Upregulation of stromal cell-derived factor 1 (SDF-1) is associated with macrophage infiltration in renal ischemia-reperfusion injury.

BackgroundStromal cell-derived factor-1(SDF-1) is a chemotactic and angiogenic factor that mediates the repair of various tissues. As macrophages are important contributors to ischemic kidney injury, we examine the role of SDF-1 in a rodent model of ischemia-reperfusion (I/R) injury.MethodsMale wild-type (WT) (C57BL/6) mice were subjected to bilateral I/R injury or sham operation in the presence or absence of macrophage depletion (liposomal clodronate [0.2 ml/20–25 g body weight i.p.]). Macrophage accumulation was assessed by immunohistochemistry. Tissue levels of SDF-1 (ELISA) and SDF-1 mRNA expression (real-time PCR) were measured. The cellular location of SDF-1 was assessed using immunohistochemical staining.ResultsImmunofluorescence staining of renal tissue sections confirmed macrophage depletion by liposomal clodronate. SDF-1 production was elevated in response to I/R injury and was significantly increased upon macrophage depletion. SDF-1 positive cells initially appeared initially in the cortex, and subsequently diffused to the outer medulla after I/R injury.ConclusionsOur study demonstrates that SDF-1 is significantly upregulated during renal I/R. We hypothesize that SDF-1 upregulation may be an important macrophage effector mechanism during I/R injury.

Amelioration of Murine Schistosoma mansoni Induced Liver Fibrosis by Mesenchymal Stem Cells

Schistosomiasis is a common chronic helminthic infection of the liver that causes hepatic fibrosis and portal hypertension,contributing to the death of over half a million people a year. Infusion of autologous bone marrow cells into patients with hepatic cirrhosis has been reported to ameliorate symptoms of portal hypertension and improve liver function, either by conversion of the infused mesenchymal stem cells (MSCs) to hepatocytes or by modulating of the hepatic fibrosis. Here,we have investigated the antifibrotic effect of mesenchymal stem cells (MSCs) using S. mansoni-induced liver fibrosis in mice, which causes an intense, stable fibrosis. MSCs derived from bone marrow of male mice were then infused intravenously into female mice that had received intraperitoneal injection of S.mansoni cercariae. Mice were divided into 4 groups: Untreated control; MSCs infusion only; Schistosomiasis only; and Schistosomiasis plus MSCs infusion. Serum alanine aminotransferase (ALT) and liver histopathology were evaluated. Expression of the collagen gene (type I),transforming growth factor (TGF-β), matrix metalloproteinase (MMP2), tissue inhibitor of metalloproteinase (TIMP-1),stromal cell-derived factor-1(SDF-1) and its receptor (CXCR4) were analyzed. MSC infusion resulted in significant decrease in liver collagen and TGF-β gene expression in the Schistosomiasis mice. The ratio of MMP-2 to TIMP-1 expression increased. SDF-1 and CXCR4 mRNA expression also increased. There was overall improvement of liver histology and a statistically significant reduction of serum ALT level. MSCs infusion ameliorated S. mansoni-induced liver fibrosis, probably by modulating the relative expression of MMP and TIMP. The findings support the hypothesis that MSCs participate in liver regeneration and functional improvement by reducing liver fibrosis.

Anti-angiogenesis effect of AMD3100 in oxygen-induced retinopathy mice

To observe the inhibition of neovascularisation in the oxygen-induced retinopathy (OIR) mice by a stromal cell-derived factor 1 (SDF-1) antagonist. Experimental study. Fifty-eight 7-day-old C57BL/6 mice were divided into 3 groups randomly, the control group (n = 17), the test group (n = 17) and the medication group (n = 24). According to the dosage of AMD3100, the medication group (n = 24) were divided into low dose group, high dose group, low dose control group, and high dose control group (each group n = 6). Each group (19-day-old) was sacrificed to perform ADPase staining, paraffin sections and immunohistochemical staining (anti-VEGF and anti-SDF-1). The average positive staining area percentage (APSAP) was measured as the outcomes and processed with the Students' t-test. Real-time PCR showed expression of both VEGF mRNA (0.080 ± 0.022 vs. 0.123 ± 0.032) and SDF-1 mRNA (0.731 ± 0.099 vs.0.544 ± 0.108) in retinas from the control group and test group, respectively. The expression of these factors in the test group was significantly higher (t = 2.488, P = 0.038;t = 2.864, P = 0.021). The number of neovascular endothelial nuclear that broke through the retinal internal limiting membrane in the paraffin section in the high dose group and the low dose group was significantly less than that in the self-control group (t = -9.507, P = 0.000; t = -10.761, P = 0.000). The appearance of ADPase staining sections in the medication group was more similar to the simple control group than that of the test group. Immunohistochemical staining sections showed that VEGF and SDF-1 expressed in neuroepithelial cells in each group. APSAP in the high dose group and the low dose group was significantly lower than that in the self-control group (VEGF: t = -7.249, P = 0.000; t = -9.02, P = 0.000; SDF-1: t = -5.246, P = 0.000; t = -5.216, P = 0.000). These results indicate that AMD3100 block the SDF-1 receptor to reduce the effect of SDF-1, decrease the production of VEGF protein and inhibite neovascularization.

Role of Stromal Cell‐Derived Factor‐1 Expression in the Injured Mouse Auditory Nerve

The degeneration of hair cells and spiral ganglion neurons (SGNs) is an important pathologic process in the development of sensorineural hearing loss. In a murine model, predictable and reproducible damage to SGNs occurs through the application of ouabain to the round window. Recent evidence has shown that the chemokine stromal cell-derived factor-1 (SDF-1) is a potent chemoattractant of hematopoietic stem cells (HSCs) and provides trophic support to injured tissues during development and maturation. The hypothesis for the current study is that expression of SDF-1 plays an important role in protecting SGNs and preventing further degeneration in the setting of cochlear injury. Prospective, controlled. Academic research laboratory. Auditory brainstem response (ABR) and the expression of SDF-1 mRNA and protein were examined 1, 3, 7, 14, and 30 days after application of ouabain in 35 adult mice. Following ouabain application, real-time reverse-transcription polymerase chain reaction for SDF demonstrates increased mRNA expression following ouabain injury in nontransplanted mice. A significant increase in SDF protein expression was also observed using immunolabeling techniques and Western blot analysis. SDF-1 expression is increased in the auditory nerve following cochlear injury. Further knowledge about the cochlear microenvironment, including SDF-1, is critical to maximizing HSC engraftment in the injured cochlea and providing a therapeutic option for sensorineural hearing loss.

Vascular Gene Transfer of SDF-1 Promotes Endothelial Progenitor Cell Engraftment and Enhances Angiogenesis in Ischemic Muscle

Gene therapy approaches to enhance endothelial progenitor cell (EPC) homing may augment cell engraftment to ischemic tissue and lead to a greater therapeutic response. Therefore, we assessed the effects of ultrasound-mediated (UM) transfection of the chemokine stromal cell-derived factor-1 (SDF-1) on homing and engraftment of intravenously administered EPCs and the subsequent angiogenic response in chronically ischemic skeletal muscle. Bone marrow-derived EPCs were isolated from donor Fisher 344 rats, cultured and labeled in preparation for injection into recipient animals via a jugular vein. Using a model of chronic hindlimb ischemia in rats, we demonstrated that UM destruction of intravenous carrier microbubbles loaded with SDF-1 plasmid DNA resulted in targeted transfection of the vascular endothelium within ischemic muscle and greater local engraftment of EPCs. The combination of SDF-1gene therapy and EPCs lead to the greatest increase in tissue perfusion and microvascular density within ischemic muscle, compared to no treatment or either monotherapy alone. Our results demonstrate that UM transfection of SDF-1 improves EPC targeting to chronically ischemic tissue, enhancing vascular engraftment and leading to a more robust neovascularization response.

Cancer-associated fibroblasts correlate with poor prognosis in rectal cancer after chemoradiotherapy

Cancer-associated fibroblasts (CAFs) in the stroma play an important role in influencing the proliferation, invasion and metastasis of cancer cells. Fibroblast activation protein-α (FAP-α) is known as a marker of CAFs, while stromal cell-derived factor-1 (SDF-1) is primarily expressed by CAFs. Herein, we investigated whether the expression levels of these genes are associated with clinical outcome after pre-operative chemoradiotherapy (CRT) in rectal cancer patients. We obtained total RNA from residual cancer stroma using microdissection from a total of 52 rectal cancer specimens from patients who underwent pre-operative CRT, we performed transcriptional analyses, and the serum protein concentrations in 40 matched microdissected specimens were measured by enzyme-linked immunosorbent assay. Additionally, we sought to clarify the location of FAP-α and SDF-1 expression using immunohistochemical staining. Of the 52 patients, 15.6 and 36.8% showed detectable FAP-α and SDF-1 mRNA expression, respectively. A significant correlation was observed between stromal FAP-α and SDF-1 mRNA levels. Moreover, there was a significant correlation between stromal SDF-1 gene expression levels and serum protein levels. Patients who developed distant recurrences after CRT had positive expression of both genes (P<0.05). The positive expression of both genes was also associated with poor probability of recurrence-free and overall survival (P<0.05). Patients with elevated serum SDF-1 levels had equally poor overall survival as those with positive stromal SDF-1 gene expression (P<0.05). In immunohistochemistry, both FAP-α and SDF-1 expression was observed in certain activated fibroblasts. In conclusion, FAP-α and SDF-1 expression was shown to be involved in tumor re-growth and recurrence in rectal cancer patients treated with pre-operative CRT.

CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma

To investigate the role of CXC chemokine receptor-4 (CXCR4) and stromal cell-derived factor-1 (SDF-1) in lymph node metastasis of gastric carcinoma. In 40 cases of gastric cancer, expression of CXCR4 mRNA in cancer and normal mucous membrane and SDF-1 mRNA in lymph nodes around the stomach was detected using quantitative polymerase chain reaction (PCR) (TaqMan) and immunohistochemistry assay. SGC-7901 and MGC80-3 cancer cells were used to investigate the effect of SDF-1 on cell proliferation and migration. Quantitative reverse transcription PCR and immunohistochemistry revealed that the expression level of CXCR4 in gastric cancer was significantly higher than that in normal mucous membrane (1.6244 ± 1.3801 vs 1.0715 ± 0.5243, P < 0.05). The expression level of CXCR4 mRNA in gastric cancer with lymph node metastasis was also significantly higher than that without lymph node metastasis (0.823 ± 0.551 vs 0.392 ± 0.338, P < 0.05). CXCR4 expression was significantly related to poorly differentiated, high tumor stage and lymph node metastasis. Significant differences in the expression level of SDF-1 mRNA were found between lymph nodes in metastatic gastric cancer and normal nodes (0.5432 ± 0.4907 vs 0.2640 ± 0.2601, P < 0.05). The positive expression of SDF-1 mRNA in lymph nodes of metastatic gastric cancer was consistent with the positive expression of CXCR4 mRNA in gastric cancer (r = 0.776, P < 0.01). Additionally, human gastric cancer cell lines expressed CXCR4 and showed vigorous proliferation and migratory responses to SDF-1. AMD3100 (a specific CXCR4 antagonist) was also found to effectively reduce the migration of gastric cancer cells. The CXCR4/SDF-1 axis is involved in the lymph node metastasis of gastric cancer. CXCR4 is considered as a potential therapeutic target in the treatment of gastric cancer.

Inhibitory effect of CXC chemokine receptor 4 antagonist AMD3100 on bleomycin induced murine pulmonary fibrosis

CXC chemokine receptor 4 (CXCR4), which binds the stromal cell-derived factor-1 (SDF-1), has been shown to play a critical role in mobilizing the bone marrow (BM)-derived stem cells and inflammatory cells. We studied the effects of AMD3100, CXCR4 antagonist, on a murine bleomycin-induced pulmonary fibrosis model. Treatment of mice with AMD3100 in bleomycin-treated mice resulted in the decrease of SDF-1 in bronchoalveolar lavage (BAL) fluids at an early stage and was followed by the decrease of fibrocytes in the lung. AMD3100 treatment decreased the SDF-1 mRNA expression, fibrocyte numbers in the lung at an early stage (day 3) and CXCR4 expression at the later stage (day 7 and 21) after bleomycin injury. The collagen content and pulmonary fibrosis were significantly attenuated by AMD3100 treatment in later stage of bleomycin injury. AMD3100 treatment also decreased the murine mesenchymal and hematopoietic stem cell chemotaxis when either in the stimulation with bleomycin treated lung lysates or SDF-1 in vitro. In BM stem cell experiments, the phosphorylation of p38 MAPK which was induced by SDF-1 was significantly blocked by addition of AMD3100. Our data suggest that AMD3100 might be effective in preventing the pulmonary fibrosis by inhibiting the fibrocyte mobilization to the injured lung via blocking the SDF-1/CXCR4 axis.

Stromal cell-derived factor-1 (SDF-1) expression in the oral squamous cell carcinoma

Purpose: Chemokines are structurally related, small polypeptide signaling molecules that bind to and activate a family of transmembrane G protein- coupled receptors, the chemokine receptors. Recently, interaction between the chemokine receptor CXCR4 and its ligand, stromal cell-derived factor 1 (SDF-1 or CXCL12), has been found to play an important role in tumorigenicity, proliferation, metastasis and angiogenesis in ma ny cancers such as lung cancer, breast cancer, melanoma, glioblastoma, pancreatic cancer and cholangiocarcinoma. Hence, the goal of this study is to identify the correlation of clinicopathological factors and the up-regulation of SDF-1 expr ession in oral squamous cell carcinoma. Material and methods: We studied the immunohistochemical staining of SDF-1, quantitative RT-PCR (qRT-PCR) of SDF-1 gene in 20 specimens of 20 patients with oral squamous cell carcinoma. Results: 1. In the immunohistochemical study of poor differentiated and invasive oral squamous cell carcinoma, the high level staining o f SDF-1 was observed. And the correlation between immunohistochemical SDF-1 expression and tumor nodes metastases (TNM) classification of specimens was significant.(x 2 test, P < 0.05) 2. In the SDF-1 gene qRT-PCR analysis, SDF-1 expression was more in tumor tissue than in carcinoma in situ tissue. Paired-sampl es analysis deter- mined the difference of SDF-1 mRNA expression level between the cancer tissue and the carcinoma in situ tissue.(Student ' s t-test, P < 0.05) Conclusion: These findings suggest that up-regulation of the SDF-1 may play a role in progression and invasion of oral squamous cell carcin oma.

Expression pattern of stromal cell-derived factor-1 chemokine in invasive breast cancer is correlated with estrogen receptor status and patient prognosis

Chemokine receptor CXCR4 is known to be crucially involved in tumor progression, but the role of its ligand, stromal cell-derived factor-1 (SDF-1), remains unclear. The present study was conducted to clarify the clinicopathological and prognostic impact of SDF-1 expression in invasive breast cancers. Expression of SDF-1 mRNA and protein was examined in five breast cancer cell lines with or without estradiol treatment. In 52 surgically resected breast cancers, the level of SDF-1 mRNA in frozen samples and the pattern of SDF-1 protein immunoreactivity in formalin-fixed paraffin-embedded tissue sections were compared. In another cohort of 223 breast cancers, the correlation between SDF-1 immunoreactivity and clinicopathological parameters was examined using a tissue microarray. Estradiol treatment markedly increased the expression of SDF-1 mRNA and protein in the estrogen receptor (ER)-positive cell lines, MCF-7 and T47D. Among the 52 resected breast cancers, those with a cytoplasmic-dominant pattern of SDF-1 expression showed higher SDF-1 mRNA levels (median 27.4) than those with a membrane-dominant or negative pattern (median 13.6, P = 0.0017). Accordingly, the cytoplasmic-dominant pattern was defined as "high SDF-1 expression," and other patterns were defined as "low SDF-1 expression." Among the cohort of 223 tumors, "high SDF-1 expression" was detected in 158 (70.9%) and was significantly correlated with ER positivity (P < 0.0001), HER2 negativity (P = 0.021), and lower grade (P < 0.0001). Univariate analysis demonstrated that "high SDF-1 expression" was a significant indicator of better clinical outcome in both the entire patient cohort (P = 0.017) and the 133 patients with ER-positive tumors (P = 0.036), but not in the 90 patients with ER-negative tumors. Multivariate analysis showed that SDF-1 status was an independent factor related to overall survival in patients with ER-positive tumors (P = 0.046). SDF-1 status is a significant prognostic factor and may be clinically useful for assigning adjuvant therapy to patients with ER-positive invasive breast cancers.

Chemokines and pituitary functions

Event Abstract Back to Event Chemokines and pituitary functions Tullio Florio1* 1 University of Genova Viale Benedetto XV, Department of Oncology Biology and Genetics, Italy Stromal cell-Derived Factor 1 (SDF1) and its receptor, CXCR4, were reported to play a regulatory roles in extra-lymphoid cells and in particular in the Central Nervous System. SDF1 and CXCR4 mRNA expression was identified in many tumor histotypes and were involved in proliferation, migration, metastasization and neoangiogenesis. To date no data are available on the role of SDF1/CXCR4 in normal pituitary function and in the genesis or deveoplment of pituitary adenomas. We previously reported SDF1 to induce GH and prolactin secretion and proliferation in GH4C1 rat pituitary adenoma cells, through the activation of multiple intracellular systems. In this study, we analyzed the expression and the biological significance of the SDF1/CXCR4 ligand/receptor pair in human pituitary adenomas and in human normal pituitaries.The expression of SDF1 and CXCR4 in 56 human pituitary adenomas (25 GH-secreting, GHoma, and 31 clinically non-functioning, NFPA) and 4 human normal pituitaries was determined by RT-PCR, immunohistochemistry and confocal immunofluorescence. The proliferative effect of SDF1 was evaluated in 8 fibroblast-free human pituitary adenoma cell cultures.CXCR4 mRNA was expressed in 92% of GHomas and 81% of NFPAs, while SDF1 was identified in 63% and 78% of GHomas and NFPAs, respectively, being the co-expression of both molecules the most common event. Immunostaining for CXCR4 and SDF1 demonstrated a strong homogenous labeling in all tumoral cells in both GHomas and NFPAs. In normal tissues, CXCR4 and SDF1 were express only in a subset of anterior pituitary cells, with a lower expression of SDF1 compared to its cognate receptor. CXCR4 and SDF1 were not confined to a specific cell population in the anterior pituitary, but co-localized with discrete subpopulations of GH, prolactin and ACTH-secreting cells. Conversely, most of the SDF1-containing cells expressed CXCR4. No colocalization of CXCR4 and SDF1 was observed within the tumor mass with CD34 (endothelial cells) or in normal pituitary with S100 (folliculostellate cells) or CD34, indicating a selective expression with pituiticites. In 6 out of 8 pituitary adenoma primary cultures, SDF1 induced a statistically significant increase in DNA synthesis that was prevented by the treatment with the CXCR4 antagonist AMD3100 or somatostatin.CXCR4 and SDF1 are over-expressed in human pituitary adenomas and CXCR4 activation may contribute to pituitary cell proliferation and, possibly, to adenoma development in humans. Conference: 3rd Mediterranean Conference of Neuroscience , Alexandria, Egypt, 13 Dec - 16 Dec, 2009. Presentation Type: Oral Presentation Topic: Symposium 17 – Chemokines: new modulators in brain and pituitary functions Citation: Florio T (2009). Chemokines and pituitary functions. Front. Neurosci. Conference Abstract: 3rd Mediterranean Conference of Neuroscience . doi: 10.3389/conf.neuro.01.2009.16.071 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 20 Nov 2009; Published Online: 20 Nov 2009. * Correspondence: Tullio Florio, University of Genova Viale Benedetto XV, Department of Oncology Biology and Genetics, Genova, Italy, tullio.florio@unige.it Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tullio Florio Google Tullio Florio Google Scholar Tullio Florio PubMed Tullio Florio Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Upregulation of stromal cell–derived factor 1 (SDF-1) expression in microvasculature endothelial cells in retinal ischemia-reperfusion injury

Stromal cell-derived factor 1 (SDF-1) is a potent chemotactic and angiogenic factor that has been proposed to play a role in the development of neovascularization. In this study, we explored the expression of SDF-1 in a rat model of retinal ischemia-reperfusion injury and investigated the possible role of retinal microvasculature endothelium cells in generation of this chemokine. Expression patterns of SDF-1 were studied in retina suffering ischemia-reperfusion insult in Sprague-Dawley rats by elevating the intraocular pressure to 110 mm for 60 minutes. The relative level of SDF-1 mRNA in retinas following 6, 12 and 24 hours reperfusion was determined by semi-quantitative RT-PCR. Immunohistochemical methods were used to detect specific lesions expressing SDF-1. The gene expression of SDF-1 in cultured human retinal microvasculature endothelial cells (HRMEC) under hypoxia conditions was assessed by semi-quantitative RT-PCR. The SDF-1 protein was analyzed by immunocytochemistry and fluorescence-activated cell sorting. Upregulation of SDF-1 mRNA (at 6, 12, and 24 hours of reperfusion) was observed, with the expression peak occurring at 12 hours. SDF-1 positive cells appeared initially around the retinal vessels,which diffused into the inner retinal layers. Hypoxia enhanced the expression of HIF-1 and SDF-1 mRNA in HRMEC. The production of SDF-1 protein by HRMEC was increased up to 320% after 6 hours of hypoxia, as demonstrated by fluorescence-activated cell sorting. The results of our study indicate that endogenous SDF-1 is up-regulated in retinal microvasculature suffering ischemia insult, and that microvasculature endothelial cells are potential contributors for generation of SDF-1 in ischemic retina.

Effect of chemokine stromal cell derived factor-1 and its receptor CXCR4 on the peritoneal carcinometastasis of gastric cancer

To investigate the effect of chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 on the peritoneal carcinometastasis of gastric cancer. Human gastric cancer cells of the lie NUGC4 and mesothelial cells of the line HMrSV were cultured. RT-PCR was used to detect the expression of CXCR4 and SDF-1 mRNA in the NUGC4 and HMrSV cells. The proliferation of NUGC4 cells was detected by MTT method. In mesothelial cell adhesion teat NUGC4 cells were cultured with confluent HMrSV mesothelial cells in 24-well plate and then divided into 3 groups: chemokine group, added with SDF-1, antibody blocking group, in which the NUGC4 cells were pre-incubated with CXCR4 monoclonal antibody for 2 h and then SDF-1 was added, and control group added with only culture fluid. Microscopy was used to calculate the number of gastric cancer cells adhered with mesothelial cells. In the mesothelial cell migration test HMrSV cells were put in the upper chamber of a Transwell chamber so as to cover the infiltration membrane. This Transwell chamber was put into a culture plate, NUGC4 cells, divided into 3 groups as mentioned above were put into the upper chamber, 24 h later HE staining and microscopy were performed to calculate the number of the NUGC4 cells that penetrated the membrane. BALB/c nu/nu female nude mice underwent intraperitoneal injection of NUGC4 cells, and then with PBS or AMD3100, small molecular specific antagonist, one day after the cancer cell injection once a day for 2 weeks. Then the mice were killed to observe the intraperitoneal tumorigenesis. CXCR4 mRNA was highly expressed in the NUGC4 cells but only very weekly expressed in the HMrSV cells. SDF-1 mRNA expression was seen in the HMrSV cells but in the NUGC4 cells. Anti-CXCR4 monoclonal antibody (McAb) inhibited the proliferation of NUGC4 cells significantly (P < 0.05). In mesothelial cell adhesion test, the number of the NUGC4 cells adhered with HMrSV cells after SDF-1 stimulation was 84.4 +/- 21.2, significantly higher than that of the control group (43.6 +/- 12.4, P < 0.05). The number of migrating NUGC4 cells in the chemokine group was 170.8 +/- 24.2, significantly higher than hat of the control group (102.8 +/- 18.2, P < 0.05); and the number of migrating NUGC4 cells in the antibody blocking group was 114.7 +/- 20.3, significantly lower than that of the chemokine group (P < 0.05). The survival time of the mice injected with both NUGC4 cells and AMD3100 was (43.8 +/- 2.8) days, significantly longer than that of the control group [(28.2 +/- 2.5) days, P < 0.01]. The tumor number of the AMD3100 group was (64.6 +/- 8.2), significantly lower than that of the control group [(103 +/- 12.4), P < 0.01]. SDF-1 and its receptor CXCR4 play an important role in the development of peritoneal carcinometastasis from gastric cancer. Interfering with the SDF-1/CXCR4 biological axis may become a potential strategy in the prevention and treatment of peritoneal carcinometastasis from gastric cancer.

Stromal Cell-Derived Factor-1 Promotes Myeloma Cell Growth in Both Autocrine and Paracrine Manners

������������������������������������������������������������������������������������������������������������������������������ ������������������������������������������������������������������������������������������ Background: The question as to whether stromal cell-derived factor-1 (SDF-1) stimulated myeloma cell growth has been controversial. We explored the possibility that SDF-1 may function as an autocrine growth factor of myeloma cells. Methods: CD138+ primary bone marrow myeloma cells and myeloma cell lines (RPMI8226, U266, and ARH77) were used. Chemotaxis in response to SDF-1 of the cells was analyzed using TranswellsTM . Cell proliferation was measured by a colorimetric assay. SDF-1 mRNA expression was analyzed by RT-PCR (reverse-transcription-polymerase chain reaction). SDF-1 and interleukin-6 (IL-6) receptor expression as well as signaling molecule phosphorylation levels, were examined using Western blot analysis. Concentrations of SDF-1 in the cell culture supernatants were measured by ELISA assay. Results: SDF-1 alone had no discernible effect on the proliferation of CD138+ primary myeloma cells or myeloma cell lines. In contrast, SDF-1 significantly enhanced IL-6-induced proliferation of these cells. SDF-1 up-regulated the expression of IL-6 receptor and enhanced phosphorylation of AKT in an additive manner with IL-6. Co-culture of the myeloma cells with umbilical vein endothelial cells over-expressing the SDF-1 gene revealed that SDF-1 played an important role in not only the migration of the cells underneath the stromal cells but also the proliferation of the cells in contact with stromal cells. All the myeloma cell lines expressed SDF-1 mRNA, and SDF-1 was detected in the culture supernatants of the cells. The G protein-coupled receptor inhibitor, pertussis toxin, inhibited the proliferation of these cells in suspension cultures. Conclusion: SDF-1, most likely in concert with IL-6, enhanced the proliferation of myeloma cells in a both paracrine and autocrine manner. (Korean J Hematol 2008;43:127-137.)