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Objective:To explore the correlation of c-MET and CXCR4 proteins and microvessel density (MVD) with liver metastasis in colorectal cancer tissues.Methods:A total of 40 colorectal cancer tissue samples and 10 paracancerous (5 cm from the edge of the tumor) normal colorectal tissue samples were collected from March 2015 to December 2020 in Shanxi Traditional Chinese Medical Hospital. Among 40 patients with colorectal cancer, 15 patients had liver metastasis. Immunohistochemistry was used to detect c-MET protein, CXCR4 protein and CD34-labeled MVD in various tissues, and the relationships between them and liver metastasis and between the three were analyzed.Results:The positive rates of c-MET protein [72.5% (29/40) vs. 30.0% (3/10)], CXCR4 protein [47.5% (19/40) vs. 10.0% (1/10)] and MVD (20.1±5.2 vs. 11.5±4.3) in colorectal cancer tissues were higher than those in paracancerous tissues, and the differences were statistically significant (all P < 0.05). The positive rates of c-MET protein [86.7% (13/15) vs. 64.0% (16/25)] and CXCR4 protein [66.7% (10/15) vs. 36.0% (9/25)] in colorectal cancer liver metastasis group were significantly higher than those in non-liver metastasis group, and the differences were statistically significant (both P < 0.05). MVD in colorectal cancer liver metastasis group was significantly higher than that in non-liver metastasis group (21.5±5.3 vs. 12.4±5.7), and the difference was statistically significant ( P < 0.05). In colorectal cancer tissues, c-MET protein expression was positively correlated with CXCR4 protein expression ( r = 0.568, P < 0.05), and MVD in c-MET-positive patients or CXCR4-positive patients was higher than that in negative ones (both P < 0.05). Conclusions:The c-MET protein, CXCR4 protein and MVD may play important roles in the liver metastasis of colorectal cancer. The three indicators can provide a certain reference for the early diagnosis and prognosis prediction of liver metastasis of colorectal cancer.
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Objective:To evaluate the role of CXC chemokine receptor 6 (CXCR6)-mediated activation of natural killer T (NKT) cells in renal fibrosis following acute kidney injury (AKI) in mice.Methods:Eighteen male wild-type C57BL/6 mice and 18 CXCR6 knockout C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 6 groups ( n=6 each) using a random number table method: wild-type mouse control group (group WT-CON), CXCR6 knockout mouse control group (group CXCR6 -/--CON), wild-type mouse with AKI group (group WT-AKI), CXCR6 knockout mouse with AKI group (group CXCR6 -/--AKI), wild-type mouse with AKI + NKT cell adoptive transfer group (group WT-AKI-NKT) and CXCR6 knockout mouse with AKI + NKT cell adoptive transfer group (group CXCR6 -/--AKI-NKT). Folic acid 250 mg/kg was intraperitoneally injected to establish the model of renal fibrosis in mice with AKI.NKT cellsuspension 250 μl(1×10 6 cells) was injected through the tail vein on the 4th and 9th days after folic acid injection in group WT-AKI-NKT and group CXCR6 -/--AKI-NKT, respectively.Blood samples were taken from orbital at day 14 after folic acid injection for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr). The animals were sacrificed, and renal tissues were obtained for observation of the area of renal fibrosis (by Sirius red staining) and renal injury (using H&E staining) which was scored and for determination of the proportion of CD1d Tetramer+ cells (by flow cytometry), the number of CD206 and α-smooth muscle actin (α-SMA) double positive (CD206 + -α-SMA + ) cells (by immunofluorescence) and expression of interleukin (IL)-4 and IL-13 mRNA (by real-time polymerase chain reaction). Results:Compared with group WT-CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI and WT-AKI-NKT ( P<0.05). Compared with group WT-AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI-NKT ( P<0.05), and the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly decreased, and the expression of IL-4 and IL-13 mRNA was down-regulated in group CXCR6 -/--AKI ( P<0.05). Compared with group CXCR6 -/--CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased in group CXCR6 -/--AKI and group CXCR6 -/--AKI-NKT ( P<0.05). Compared with group CXCR6 -/--AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group CXCR6 -/--AKI-NKT ( P<0.05). Conclusion:CXCR6-mediated activation of NKT cells is involved in renal fibrosis following AKI in mice, and the mechanism may be related to promoting Th2 cytokine-mediated M2 macrophage-myofibroblast transformation.
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Objective To investigate the clinical application of chemokine receptor 4 (CXCR4)-targeted PET/CT imaging in breast cancer using 68Ga-1,4,7-triazacyclononane-1,4,7-triacetic acid-TN14003 (NOTA-NFB) and the correlation between 68Ga-NOTA-NFB uptake and pathology.Methods From June 2014 to December 2014,11 female patients (age range:38-68 years) with non-specific invasive breast cancer were recruited in this study.All patients underwent neoadjuvant chemotherapy before surgery.68GaNOTA-NFB and 18F-fluorodeoxyglucose (FDG) PET/CT imaging were performed before the chemotherapy.Three patients also underwent 68Ga-NOTA-NFB PET/CT imaging after the fourth cycle of chemotherapy.The region of interest (ROI) method was used to measure the maximum standardized value (SUVmax) and tumor/non-tumor (T/NT) ratio was calculated.Paired t test and Spearman correlation analysis were used for statistical analysis.Results The SUVmax values of primary lesions were 3.78±2.03 and 8.11±5.14 (t=-3.01,P＜0.05) respectively in 68Ga-NOTA-NFB imaging and 18F-FDG imaging.The T/NT ratios for primary lesions were not significantly different between the two imaging methods (9.36±7.81 vs 15.62±14.51;t=-1.63,P＞0.05).In the metastatic lymph nodes,SUVmax values were not significantly different between 68Ga-NOTA-NFB imaging and 18F-FDG imaging (t=-2.02,P＞0.05),but T/NT ratios were significantly different (t=-2.43,P＜0.05).After neoadjuvant chemotherapy,T/NT ratios were decreased in the 3 patients.Correlation was not found between T/NT in 68Ga-NOTA-NFB imaging and Ki-67,but the P value was close to 0.05 (rs =0.600,P=0.051).Conclusion 68Ga-NOTA-NFB PET/CT can be used as a new CXCR4-targered imaging in diagnosis of breast cancer,and it may be beneficial to evaluate the effect of neoadjuvant chemotherapy.
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Objective To detect the expression and clinical significance of stro-mal cell derived factor-1 (SDF-1) and CXC chemokine receptor 4 (CXCR4) in peripheral blood of patients with hepatitis C cirrhosis.Methods From January 2015 to January 2018,120 patients with hepatitis C cirrhosis in our hospital were selected as the subjects,including 60 patients with compensated hepatitis C cirrhosis and 60 patients with decompensated hepatitis C cirrhosis,and 60 hepatitis C patients without cirrhosis in the same period were selected as the control group.Enzyme linked immunosorbent assay (ELISA) was used to detect the expression levels of SDF-1 and CXCR4 in peripheral blood;the correlation between SDF-1 and CXCR4 expressions in compensated cirrhosis and decompensated cirrhosis patients was analyzed.Results The expression levels of SDF-1 and CXCR4 in peripheral blood samples of patients with decompensated cirrhosis of hepatitis C were significantly higher than those of patients with compensated cirrhosis of hepatitis C (P ＜ 0.05);the expression levels of SDF-1 and CXCR4 in peripheral blood samples of patients with compensated and decompensated cirrhosis of hepatitis C were significantly higher than those of control group (P ＜ 0.05);the expressions of SDF-1 and CXCR4 in peripheral blood of patients with compensated and decompensated cirrhosis of hepatitis C were positively correlated (r =0.684,P ＜ 0.05,r =0.765,P ＜ 0.05).Albumin (AlB) level in peripheral blood of cirrhosis group was significantly lower than that of control group (P ＜ 0.05);alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBIL),while fasting insulin (FINs) and interleukin (IL)-6 in cirrhosis group were higher than those of control group (P ＜ 0.05);SDF-1 level and CXCR4 level were positively correlated with AlB,FINs and IL-6 (P ＜ 0.05);multiple regression analysis showed that FINs,IL-6,SDF-1 and CXCR4 were risk factors for hepatitis C cirrhosis.Conclusions The levels of SDF-1 and CXCR4 in peripheral blood of patients with hepatitis C cirrhosis increased,and the levels of SDF-1 and CXCR4 in peripheral blood were positively correlated,suggesting that they may be involved in the regulation of the occurrence and development of hepatitis C cirrhosis.
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Objective To observe the impairment effect of retinol binding protein 4(RBP4) on neurocognitive function in diabetic nephropathy(DN) patients with silent cerebral infarction(SCI) and to explore its mechanism.Methods Sixty patients with newly diagnosed DN and 30 healthy volunteers were selected as the study subjects and the DN cases were divided into the complicating SCI group(SCI,n=30) and non-complicating SCI group(NSCI,n=30) according to the imaging results.The degrees of neurological function deficit and Montreal cognitive assessment(MoCA) were evaluated.Serum RBP4 level was determined by ELISA and expressions of Lp-PLA2 and C-X-C chemokine receptor type 4(CXCR4) were determined by Western blot.Results Compared with the NSCI group,the neurocognitive function in the SCI group was subsided,the expression levels of RBP4,Lp-PLA2 and CXCR4 were increased(P＜0.05).The RBP4 level was positively correlated with the neurocognitive function impairment in SCI patients,moreover,there existed a regression correlation between them.Conclusion Serum RBP4 may serve as the predictive factor of DN complicating SCI and is positively correlated with neurocognitive dysfunction.Lp-PLA2/CXCR4 pathway activation may be one of its pathogenesis.
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Objective To detect the expression levels of peripheral blood CXCL10 and its receptor CXCR3 and T cell subsets in of the patients with advanced vitiligo and the influence of compound Chinese medicine on it.Methods Flow cytometry was used to detect the cellular proportions of peripheral blood T cell subsets,ELISA was employed to quantify serum CXCL10 and CXCR3 expression levels before and after treatment.Results After 1 month of taking Chinese medicine,the proportions of CD3+ CD4+ cells and CD3+ CD8+ cells were increased compared before treatment(P＜0.05).The expression level of peripheral serum CXCL10 before treatment was significantly increased compare with the healthy control group(P＜0.01),and the CXCL10 level after treatment was decreased significantly compared with that before treatment(P＜0.05).The expression level of peripheral serum CXCR3 was significantly increased compared with the healthy control group(P＜0.05),while which after treatment was still significantly higher than that in the healthy control group(P＜0.05).Conclusion CXCL10,CXCR3 and T cell subsets proportion may be involved in the pathogenesis of vitiligo.The compound Chinese medicine used in this study plays the curative effect possibly by regulating T cell subsets and expression levels of CXCL10 and CXCR3.
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Background:CXCR4 is widely expressed in tumor cells and participates in tumor invasion and metastasis. RNA interference technology can effectively reduce or shut down the expression of genes and block tumor invasion and metastasis at different levels. Aims:To investigate the mRNA and protein expressions of CXCR4 and their relationship with clinicopathological features of gastric cancer,and to explore the effect of silencing CXCR4 by siRNA on biological behavior of gastric cancer. Methods:A total of 86 gastric cancer tissues and their adjacent normal mucosa were collected. mRNA and protein expressions of CXCR4 were detected by RT-PCR and Western blotting,respectively,and their relationship with clinicopathological features was analyzed. The CXCR4 RNA interference plasmid vector was constructed,and were transfected into gastric cancer cell line MKN45. Cell proliferation,migration and invasion,apoptosis were detected by MTT assay,Transwell assay and flow cytometry,respectively. Results:mRNA and protein expressions of CXCR4 in gastric cancer tissues were significantly higher than those in adjacent normal tissues (P ＜0.05),and CXCR4 expression was related to TNM staging,tumor differentiation,lymph node metastasis (P＜0.05). Compared with negative control cells, cell proliferation in siRNA transfection group at 24,48,and 72 hours were significantly decreased (P ＜0.05 ),cell migration rate and cell invasion rate were significantly decreased (P ＜0.05),and cell apoptosis rate was significantly increased (P＜0.05). Conclusions:CXCR4 plays an important role in the development of gastric cancer. The silencing CXCR4 gene by siRNA can significantly inhibit the proliferation,migration and invasion of MKN45 cells,and increase apoptosis,thereby providing a new strategy for the treatment of gastric cancer.
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Objective To investigate the mechanism and function of chemokine receptor CXCR4 and its ligand CXCL12 (CXCL12 / CXCR4) in primary hepatocellular carcinoma (PHC). Methods Western blot assay, immunohistochemistry and Real-time PCR were used to detect the protein and mRNA expressions of CXCL12/CXCR4 in 60 PHC and corresponding paracancerous tissue samples. Four kinds of hepatoma cells (Huh7, MHCC97h, HepG2 and Hep3B) and normal hepatocytes (7702) were routinely cultured, and then real-time PCR was performed to detect the mRNA expressions of CXCL12/CXCR4 in these cells to screen suitable experimental cells. CXCR4 interference plasmid (sh-CXCR4) and corresponding empty vector (sh-control) were transfected into MHCC97h to construct stable transfected cell lines. The ability of invasion, migration, and proliferation of the 2 groups of cells were detected by Tanswell invasion experiment, cell scratch test and MTT test. The stably expressed sh-control and sh-CXCR4 MHCC97h cells were taken into the subcutaneous of six nude mice, and the growth of the tumor was observed. Western blot assay was used to detect the expressions of vascular endothelial growth factor-C (VEGF-C) in sh-control and sh-CXCR4 MHCC97h cell lines and corresponding xenografts in nude mice, as the same in MHCC97h, which was transfected with CXCR overexpressed plasmid. Results (1) The results of Western blot assay, immunohistochemistry and Real-time PCR showed that the expressions of CXCL12/CXCR4 protein and mRNA were significantly higher in liver cancer tissues than those of paracancerous tissues. (2) The expression levels of CXCL12/CXCR4 mRNA were higher in Huh7, MHCC97h, HepG2 and Hep3B cells than those of 7702 cells. MHCC97h was selected as the experimental cells. The ability of invasion, migration and proliferation of MHCC97h cells transfected with sh-CXCR4 were significantly lower than those of sh-control group. Meanwhile, the growth rate of nude mice transplanted with sh-CXCR4 MHCC97h cells was also significantly lower than that of sh-control group. (3) Both in vitro and in vivo experiments showed that the expression of VEGF-C was lower in sh-CXCR4 group than that in sh-control group, and the expression of VEGF-C was obviously up-regulated after overexpression of CXCR4. Conclusion High expressions of CXCL12/CXCR4 are found in primary cancer tissues and hepatoma cells. CXCL12/CXCR4 may inhibit the proliferation and invasion of hepatoma cells by regulating the expression of VEGF-C protein.
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Objective To investigate the effects of migration and expression from chemokine receptor 4 (chemokine receptor-4,CXCR4) of rat bone marrow mesenchymal stem cells (BMSCs) which were pretreated by atorvastatin (ATV) in vitro.Methods Isolated,cultivated,identified the BMSCs,pretreated P4-P6 of BMSCs with different concentrations of ATV for 12 hours.The experimental group was divided into control group,0.1 nM/L (group 0.1 nM),1 nM/L (1 nM group),10 nM/L (10 nM group),100 nM/L (100 nM group),1 000 nM/L (1 000 nM group).The mRNA and protein of CXCR4 were determined by real time-polymerase chain reaction and Western blot.Immunofluoreseence assay were used to detect the expression levels of CXCR4.The migration ability of BMSCs were measured by transwell chamber.Results Immunofluoreseence assay showed the protein level of CXCR4 of group 1 nM and 10 nM were significantly higher than the other group.RT-PCR and Western blot showed the protein and mRNA level of CXCR4 in 10 nM was higher than that in group 1 rM.The migration ability of group 10 nM was higher than 1 nM and control group.Conclusions ATV can be dose-dependent promote expression levels of CXCR4 of BMSCs cultivated in vitro.
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Objective To evaluate the role of C-X-C chemokine receptor type 4 ( CXCR4) in the dorsal root ganglia ( DRG) in incisional pain in rats. Methods Thirty-two male Sprague-Dawley rats, aged 7-10 weeks, weighing 250-300 g, in which intrathecal catheters were successfully implanted, were divided into 4 groups (n＝8 each) using a random number table method: sham operation group (group S), CXCR4 antagonist AMD3100 plus sham operation group (group A+S), incisional pain group (group I) and CXCR4 antagonist AMD3100 plus incisional pain group (group A+I). Rats were anesthetized with sevoflu-rane. AMD3100 20 μg (in 10 μl of normal saline) was intrathecally injected, and no incision was made 30 min later in group A+S. A 1-cm longitudinal incision was made through skin, fascia and muscle of the plantar aspect of the left hindpaw in group I. AMD3100 20 μg (in 10 μl of normal saline) was intrathecally injected, and 30 min later the model of incisional pain was established in group A+I. The mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal latency ( TWL) were measured at 24 h before surgery and 2, 4, 8, 16 and 24 h after surgery. The rats were sacrificed after the last measurement of pain threshold and the DRGs of the lumbar segment (L4-6) were removed for detecting the expression of CXCR4, phosphorylated extracellular regulated protein kinase ( p-ERK) and total ERK ( t-ERK) by Western blot. Results Compared with group S, MWT was significantly decreased and TWL was shortened at T1-5in group I and group A+I, and the expression of CXCR4 and p-ERK in DRGs was significantly up-regulated (P＜0. 05), and no significant change was found in the expression of t-ERK in group I, no significant change was found in the expression of CXCR4, p-ERK and t-ERK in group A+I, and no significant change was found in the parameters mentioned above in group A+S (P＞0. 05). Compared with group I, MWT was significantly increased and TWL was prolonged at T1-5, the expression of CXCR4 and p-ERK in DRGs was down-regulated (P＜0. 05), and no significant change was found in the expression of t-ERK in group A+I (P＞0. 05). Conclusion CXCR4 in DRGs is involved in incisional pain, and the mechanism may be re-lated to activating ERK1∕2 signaling pathway in rats.
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Objective@#To evaluate the clinical characteristics, MYD88L265P mutation, CXCR4WHIM mutation and prognosis in patients with Waldenström macroglobulinemia (WM).@*Methods@#The clinical characteristics, International Prognostic Scoring System for symptomatic WM (WPSS) , and overall survival (OS) were retrospectively assayed in 93 patients with newly diagnosed WM at Peking Union Medical College Hospital during January 2000 to August 2016. The MYD88L265P mutation and CXCR4WHIM mutation were tested among 34 patients.@*Results@#The median age of the 93 patients was 64 years (range, 33-85 years) with a male-to-female ratio of 2.44. According to WPSS, we included 16 (17.2%) low-risk, 44 (47.3%) intermediate-risk and 33 (35.5%) high-risk patients. Eight patients had secondary amyloidosis. With a median follow-up of 44 (1-201) months, the median OS was 84 months. Cox regression multifactor analysis showed WPSS risk group (HR=2.342, 95% CI 1.111-4.950, P=0.025) , whether patients had secondary amyloidosis (HR=5.538, 95% CI 1.958-15.662, P=0.001) and whether patients received new drugs (HR=3.392, 95% CI 1.531-7.513, P=0.003) were independent factors associated with OS. We have investigated the presence of the MYD88L265P and CXCR4WHIM mutation in 34 patients and found that MYD88L265P mutation was occurred in 32 patients (94.1%) and CXCR4WHIM mutation was occurred in 8 patients (23.5%). Seven of 8 patients who harbored CXCR4WHIM-mutated also exhibited the MYD88L265P mutation. Patients with MYD88L265PCXCR4WHIM vs MYD88L265PCXCR4WT presented with more severe anemia, lower platelet level, higher M protein level and more hyper-viscosity syndrome.@*Conclusion@#WPSS risk group, whether patients had secondary amyloidosis or received new drugs are independent factors for OS in WM. MYD88L265P and CXCR4WHIM mutation, the most common somatic variants in WM, often occur together and impact the clinical presentation.
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Objective To detect levels of C?X?C chemokine receptor type 5 (CXCR5) and inducible costimulator(ICOS)in blister fluid of patients with bullous pemphigoid(BP), and to explore their significance in the pathogenesis of BP. Methods Blister fluid samples were collected from 15 patients with BP(experimental group)and 15 patients with second?degree burns(control group). Enzyme?linked immunosorbent assay(ELISA)was performed to detect the levels of CXCR5 and ICOS in the 2 groups. Results The level of CXCR5 was significantly higher in the experimental group than in the control group(219 ± 145.31 vs. 147 ± 23.83 ng/L, t=4.577, P 0.05). Conclusion The expression of CXCR5 may be associated with the occurrence of BP, but further researches are needed to determine the relationship between ICOS and the occurrence of BP.
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Objective To explore the mechanism of vessel endothelial dysfunction in rats under intermittent hypoxia (IH). Methods The respiratory simulation system was used to simulate IH. Sixty C57BL/6J rats (male) were randomized into control group and IH group. The rats of IH group were exposed to IH 8 hours per day for 6 weeks. The serum levels of hypoxia inducible factor (HIF)-1a and stromal cell derived factor (SDF)-1a were assessed by ELISA. The serum levels of reactive oxygen species (ROS) were detected in two groups. The serum expression of miR-199a-5p was detected by real-time fluorescent quantitative PCR in two groups. The dual luciferase report system and point mutation test were used to verify target gene for HIF-1a. Results The serum levels of HIF-1a and SDF-1a were significantly higher in IH group than those of control group (μg/L:1.60±0.02 vs. 1.19±0.02, 1 823.00±8.97 vs. 1 444.00±17.90, P<0.01). The serum level of ROS was significantly higher in IH group than that of control group (U/mL:487.66±35.73 vs. 211.57±23.82, P<0.01). The serum level of miR-199a-5p expression was significantly lower in IH group compared to that of control group (1.31±0.07 vs. 3.47± 0.17, P<0.01). The result of dual luciferase reporter gene detection confirmed that target gene of miR-199a-5p was HIF-1a. Conclusion The serum level of miR-199a-5p is decreased first due to IH, and then its target gene (HIF-1a) is increased. HIF-1a can induce the increased level of SDF-1a, and its receptor (CXCR-4 ) is also increased. Finally, HIF-1a can increase the serum level of ROS, resulting in the endothelial dysfunction.
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Objective@#To explore the mechanism of ibrutinib on drug resistance diffuse large B-cell lymphoma (DLBCL) cells.@*Methods@#DLBCL cell line was cultured with mesenchymal stem cells (MSC) , and DLBCL cells which migrated and adhered to MSC under microscope was counted. The secretion of CXCL12 by MSC were measured by ELISA. The expression of CXCR4 on DLBCL cells were measured by flow cytometry, HBL-1 cells were transfected with a CXCR4-lentivector. An Annexin Ⅴ-binding assay was used to detect the induction of apoptosis. Clonogenic growth of DLBCL cells was evaluated on MethoCult media. Ibrutinib was injected into NOD/SCID mice, tumor growth was assessed via caliper measurements every 3 days.@*Results@#MSC promoted migration and adhesion of DLBCL cells to MSC. Ibrutinib inhibited migration and adhesion of DLBCL cells to MSC in a dose-dependent manner (P<0.05) . CXCL12 secreted by MSC and CXCR4 expressed on DLBCL cells could induce each other, which upgraded the levels of secretion and expression. Ibrutinib could inhibit the secretion of CXCL12 (SUDHL10: 660 pg/ml vs 1 400 pg/ml, P=0.004; HBL-1: 720 pg/ml vs 1 490 pg/ml, P=0.018; DLBCL:850 pg/ml vs 1 450 pg/ml, P=0.004) and expression of CXCR4 (P<0.05) . When co-cultured with MSC, the ratio of HBL-1 cells apoptosis in the group of control, mitoxantrone, ibrutinib, mitoxantrone+ibrutinib were respectively 15.1%, 17.5%, 23.5%, 58.7%. After transfected with a CXCR4-lentivector and overexpressed CXCR4, the ratios of HBL-1 cells apoptosis were 14.2%, 16.1%, 22.5%, 38.3% respectively. The ratio of DLBCL cells apoptosis induced by mitoxantrone was lower when co-cultured with MSC (P<0.05) . But with the addition of ibrutinib, the ratio of apoptosis was increaed and it was similar to cultivation without MSC, which suggested ibrutinib could inhibit drug-resistance induced by MSC. But after transfected with a CXCR4-lentivector, the overexpression of CXCR4 was detected and the ratio of apoptosis was significantly lower when co-cultured with MSC which demonstrated that ibrutinib inhibited drug-resistance by inhibiting the expression of CXCR4. MSC enhanced lymphoma clonogenicity in vitro and lymphoma cell growth in vivo. The number of colonies of control, MSC, Ibrutinib, MSC+Ibrutinib were 113±5, 205±4, 62±9, 123±3 (2.5×103/well, ±s) , respectively. The tumor volume of NOD/SCID mice were respectively 6 500, 17 000, 4 000, 10 000 mm3. Ibrutinib inhibited lymphoma clonogenicity in vitro and lymphoma cell growth in vitro.@*Conclusion@#Ibrutinib targeted the CXCL12/CXCR4 axis, inhibited the expression of CXCR4 and inhibited MSC-mediated drug resistance. Ibrutinib also inhibited lymphoma clonogenicity in vitro and lymphoma cell growth in vivo. These results provided a scientific rationality for relapsed/refractory DLBCL treatment with ibrutinib.
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Background Acquired immune deficiency syndrome (AIDS) is an infectious disease caused by human immunodeficiency virus (HIV).Highly active antiretroviral therapy (HAART) is an effective treatment for AIDS,but it cannot completely eliminate the viral load in the body for the existence of HIV reservoir.Previous studies demonstrated that HIV could be detected in tears of virus load negative AIDS patients who received effective HAART,suggesting that lacrimal gland is another member of HIV reservoirs.Objective The aim of this study was to explore whether lacrimal gland has a molecular basis of HIV infection and the mechanism of lacrimal gland infection of HIV.Methods Fourteen specimens of lacrimal gland were collected during the surgery from 14 patients with lacrimal gland diseases in Peking Union Medical College Hospital from November 2013 to December 2015,including 13 non-HIV-infected patients and 1 HIV-infected patient.In 13 non-HIV infected patients,lacrimal glands prolapse was in 12 patients with the normal pathological tissue structure and dacryoadenitis was in 1 patient with the histopathological diagnosis of interstitial lymphoid tissue hyperplasia.The clinical manifestation of HIV-infected patient was dacryoadenitis with the histopathological diagnosis of interstitial lymphoid tissue hyperplasia.The paraffin sections of 12 non-HIV-infected specimens and 1 HIV-infected specimen were prepared,and the expressions of CD4,C-X-C chemokine receptor 4 (CXCR4) and C-C chemokine receptor type 5 (CCR5) in lacrimal gland specimens were detected by immunohistochemistry and verified in 1 specimen of non-HIV-infected specimen by immunofluorescence technology.Results Immunohistochemistry showed that CD4 was suspiciously positive expression in non-HIV-infected specimens with the strong background staining.CXCR4 was positively expressed in cytoplasm and nuclei of most lacrimal epithelial cells of lacrimal gland epithelial cells in each specimen,and CCR5 was focally expressed in few lacrimal gland epithelial cells in each specimen.In addition,CD4,CXCR4 and CCR5 were positively expressed in intercellular scattered lymphocytes on the specimens.Immunofluorescence assay showed that CD4,CXCR4 and CCR5 were expressed in the specimens with the red fluorescence,with the linear-and patchy-like distribution mainly in cellular membrane for CD4 or spot-like distribution for CXCR4 and CCR5 in the cytoplasm.Conclusions HIV receptor CD4 and accessory receptor CXCR4,CCR5 are positively expressed in the lacrimal gland epithelial cells,which is the molecular basis of HIV infection and become a potential HIV reservoir preventing HIV eradication.
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BACKGROUND: Tumor associated macrophages (TAMs) and CXC chemokine receptor 4 (CXCR4) have emerged as potential biomarkers in various human cancers. The aims of this study were to investigate the clinical characteristics of anaplastic thyroid cancer (ATC) patients according to the TAM numbers in the tumor tissue, and to evaluate the associations between CXCR4 expressions and macrophage densities in ATC tumor microenvironment. METHODS: Total 14 ATC samples from thyroid tissue microarray were used. Immunohistochemical staining was performed using anti-CD163 and anti-CXCR4 antibodies. According to the immunoreactivity of CD163, all subjects were divided into two groups: low-CD163 (n=8) and high-CD163 (n=6) groups. RESULTS: The mean diagnostic age was 65±7 years and the median tumor size was 4.3 cm, ranging 2.5 to 15 cm. Clinicopathological characteristics were not significantly different between low-CD163 and high-CD163 groups, while age of diagnosis was younger in high-CD163 group than that of low-CD163 group with marginal significance (56.9±5.5 years vs. 67.5±6.8 years, P=0.09). However, overall survival was significantly reduced in high-CD163 group (5.5 months [range, 1 to 10]) compared with low-CD163 groups (8.8 months [range, 6 to 121); log-rank test, P=0.0443). Moreover, high-CD163 group showed strong CXCR4 expressions in both cancer and stromal compartments, while low-CD163 group showed relatively weak, stromal-dominant CXCR4 expressions. Additionally, CD163 and CXCR4 expressions showed a strong positive correlation (γ²=0.432, P=0.013). CONCLUSION: Increased number of TAMs showed poor overall survival in ATC, suggesting TAMs are potentially a prognostic biomarker for ATC. CXCR4 expression was significantly correlated with CD163-positive TAM densities, which suggest the possible role of CXCR4 in TAM recruitments.
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Humans , Antibodies , Biomarkers , Diagnosis , Macrophages , Receptors, CXCR , Receptors, CXCR4 , Thyroid Carcinoma, Anaplastic , Thyroid Gland , Tumor MicroenvironmentABSTRACT
Background:CXC chemokine receptor type 2(CXCR2)is a member of G protein coupled receptor superfamily,and is mainly involved in the growth of tumor,angiogenesis and the pathogenesis of inflammatory diseases. Studies showed that CXCR2 was associated with the pathogenesis of inflammatory bowel disease(IBD),but the exact role has not yet been clarified. It was found that the interaction of interleukin-8(IL-8)with CXCR1 and CXCR2 played an important role in the pathogenesis of IBD. Aims:To investigate the expressions and significance of CXCR2 and IL-8 in patients with IBD. Methods:A total of 121 IBD patients in active stage from October 2013 to December 2014 at Zhongnan Hospital of Wuhan University were enrolled and assigned into Crohn’s disease(CD)group and ulcerative colitis(UC)group. Seventy healthy subjects were served as controls(HC). Expressions of IL-8 mRNA and CXCR2 mRNA in peripheral blood and intestinal mucosal tissue were determined by real-time PCR;expression of CXCR2 protein in intestinal mucosal tissue was determined by Western blotting. Results:In peripheral blood,expression of IL-8 mRNA in UC group was significantly higher than that in HC group(P = 0. 017),while expressions of CXCR2 mRNA in CD,UC and HC groups were not significantly different (P = 0. 285). In intestinal mucosal tissue,expressions of IL-8 mRNA in CD,UC and HC groups showed no significant difference(P = 0. 206),while expressions of CXCR2 mRNA in CD and UC groups were significantly higher than that in HC group(P = 0. 002;P < 0. 001),and expression of CXCR2 mRNA in UC group was significantly higher than that in CD group(P = 0. 005);expressions of CXCR2 protein in UC and CD groups were higher than that in HC group(P = 0. 049 P =0. 080). Conclusions:Expressions of CXCR2 mRNA and protein in intestinal mucosal tissue of patients with IBD, especially UC are significantly increased. Down regulation of CXCR2 expression in intestinal mucosa may provide a new target for treatment of UC. IL-8 is significantly highly expressed in peripheral blood of patients with UC,which suggests that IL-8 might be related mainly with UC.
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Background Study confirmed that the active microglia may injure retinal ganglion cells (RGCs) in retinal ischemia reperfusion injury (IRI), and increased cx3cr1 expression is an important factor in microglial activation,and thus blocking the expression of cx3cr1 can inhibit microglial activation, which may be useful in neuronal protection.Objective This study was to analyze the protective effects of cx3cr1 antibody on retinal neuron in rat eyes with IRI.Methods Ninety SD rats were divided into 4 groups according to random number table.IRI models were established by perfusing normal saline solution into the anterior chamber.The cx3cr1 antibody of 1 μl (0.2 μg/μl) was intravitreally injected in the right eyes in the normal rats or model rats as the only cx3cr1 antibody injected group and the model cx3cr1 antibody injected group,respectively,and no any drug was injected in the rats of the normal control group and model control group.Retinal sections were prepared 48 hours after modeling, and apoptosis of retinal neutron was observed under the transmission electron microscope;the morphology of retinas was exmined and the number of survival RGCs was calculated by histopathologic method.The expression of CD68 in activated retinal microglial cells was detected by immunochemistry, and the relative expression levels of cx3cr1 mRNA,tumor necrosis factor-α (TNF-ct) mRNA and interleukin-1 β (IL-1 β) mRNA in the retinas were assayed by real time quantitative PCR.Results The cell nucleus of RGCs showed the round and ellipse in shape and there were abundant organelles in the cells.The mophology of photoreceptors was normal with abundant mitochondrions.Irregular cell shape, disrupture of outer segment membranous disc, proliferative microglial cells in RGC layer were seen in the model group.However,these findings were mild in the model cx3crl antibody group.The mean number of survival RGCs was (38.100 ± 3.929), (37.200 ± 5.266), (26.700 ± 2.584) and (31.700 ± 2.946)/field in the normal control group,only cx3cr1 antibody injected group, model control group and model cx3cr1 antibody injected group,showing significant differences between the model group and the normal control group, only cx3cr1 antibody injected group or model cx3cr1 antibody injected group (t =7.492,6.125,-4.607, all at P＜0.01).The expression levels (absorbance) of CD68 in rat retinas were significantly higher in the model group than those in the normal control group, only cx3cr1 antibody injected group and model cx3cr1 antibody injected group (t =-3.397 ,P =0.008;t =-6.207 ,P =0.000;t =3.494, P =0.007).The relative expression levels of cx3cr1 mRNA, TNF-α mRNA and IL-1 β mRNA in rat retinas were raised in the model group compared with the only cx3er1 antibody injected group and model cx3cr1 antibody injected group (all at P＜0.01).No significant differences were observed in these indicators between the normal control group and the only cx3crl antibody injected group (all at P＜0.05).Conclusions Intravitreal injection of cx3cr1 antibody to neutralize cx3cr1 levels in retinas can effectively inhibit the activation of retinal microglia,decrease the release of inflammatory factors, reduce the apoptosis of RGCs and thereby protect the retinal neutrons against IRI in SD rats.Intravitreal injection of cx3cr1 is safe and feasible.
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Hepatitis C virus (HCV) infection represents an important public health problem worldwide. Eradicating HCV can finally delay or prevent the progression of HCV infection to end-stage liver diseases such as liver cirrhosis and liver cancer. Chemokine (C-X-C motif) ligand 10 (CXCL10) is a chemokine belonging to the CXC chemokine family, and it exerts its function through binding to chemokine (C-X-C motif) receptor 3 (CXCR3), playing a critical role in eradication of HCV. This article reviews the relationship of CXCL10 with the pathogenesis of HCV and the effectiveness of antiviral treatment, as well as the CXCL10 measurements. Meanwhile, this article introduces the clinical value of CXCL10 in assessing the risk of liver fibrosis and liver cancer. Further studies are needed to investigate the association between CXCL10 and liver diseases, and CXCL10 may provide a new therapeutic strategy for HCV infection.
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Objective To study the effect of Numb gene on CXCR4 expression and proliferation and migration of bladder cancer. Methods Human bladder cancer cells were divided into 3 groups:experimental group(transfected with Numb-ORF plasmid),negative-control group( transfected with blank vector),and blank-control group(no DNA transfected in). The expressions of Numb and CXCR4 were detected by real-time PCR and Western blotting respectively. Cell proliferation and migration were measured by MTS and Transwell assay respectively. Results Numb expressions in experimental group,negative-control group and blank-control group were 31. 044 ± 3. 350,4. 401 ± 0. 567 and 4. 287 ± 0. 341 respectively,with a statistical significance (F = 183. 418,P = 0. 000). CXCR4 levels in experimental group,negative-control group and blank-control group were 0. 344 ± 0. 167,0. 996 ± 0. 148 and 1. 010 ± 0. 106 respectively,with a statistical significance (F = 21. 355,P = 0. 002). In MTS assay,the absorb values in experimental group,negative-control group and blank-control group were 0. 615 ± 0. 057,0. 987 ± 0. 063 and 0. 957 ± 0. 066,with a statistical difference(F =33. 210,P = 0. 001). In Transwell assay,the numbers of migratory cells in experimental group,negative-con-trol group and blank-control group were 164. 667 ± 19. 858,670. 133 ± 38. 760 and 667. 533 ± 27. 610,with a statistical significance(F = 286. 788,P = 0. 000). Conclusion Overexpression of Numb can suppress the ability of proliferation and migration of the BIU-87 cells through down-regulation of CXCR4 expression in human bladder cancer.