Carnosic acid affects the proliferation, migration, and invasion of gastric cancer AGS cells by regulating CXCR7/CXCL12 axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨鼠尾草酸（CA）通过调节CXC基序趋化因子受体7（CXCR7）/CXC基序趋化因子配体（CXCL12）轴对胃癌AGS细胞增殖、迁移和侵袭的影响。方法：用不同浓度（0、5、10、20、40、80 µg/mL））的CA处理胃癌AGS细胞，采用CCK-8法筛选合适的CA浓度；将AGS细胞分为对照组（未经处理的AGS细胞）、CA组（20 µg/mL CA处理）、CA+siCXCR7组（转染siCXCR7+20 µg/mL CA处理）、CA+siNC组（转染siNC+20 µg/mL CA处理）、CA+vectorNC组（转染vectorNC+20 µg/mL CA处理）、CA+vectorCXCR7组（转染vectorCXCR7+20 µg/mL CA处理），采用CCK-8法检测AGS细胞增殖的变化，qPCR法检测细胞中CXCR7、CXCL12 mRNA表达水平的变化，Transwell实验检测细胞侵袭能力的变化，划痕实验检测细胞迁移能力的变化，WB法检测周期蛋白D1、Bcl-2、CXCR7、CXCL12、MMP-2蛋白表达的变化。结果：不同浓度CA均可抑制AGS细胞存活率，且浓度为20 µg/mL时，细胞存活率接近50%，故选择20 µg/mL CA用于后续研究。与对照组相比，CA组增殖率、侵袭数、迁移率、周期蛋白D1、MMP-2、Bcl-2、CXCR7、CXCL12 mRNA及蛋白表达显著降低（均P<0.05）；与CA+siNC组相比，CA+siCXCR7组增殖率、侵袭数、迁移率、周期蛋白D1、MMP-2、Bcl-2、CXCR7、CXCL12 mRNA及蛋白表达显著降低（均P<0.05）；与CA+vectorNC组相比，CA+vectorCXCR7组增殖率、侵袭数、迁移率、周期蛋白D1、MMP-2、Bcl-2、CXCR7、CXCL12 mRNA及蛋白表达显著增加（均P<0.05）。结论：CA可抑制AGS细胞增殖、迁移和侵袭，其机制可能与抑制CXCR7/CXCL12轴有关。

CENPF Promotes EMT in Non-small Cell Lung Cancer by Up-regulating ACKR3/CXCR7 / 肿瘤防治研究

Objective To investigate the relationship between the expression of CENPF in NSCLC adenocarcinoma (LUAD) and the clinical prognosis of patients and its effect on the metastasis of lung adenocarcinoma cells. Methods The expression of CENPF in LUAD and its relationship with patient prognosis were analyzed by online bioinformatics. The expression of CENPF was verified by LUAD tissue microarray immunohistochemical staining. Kaplan-Meier analysis was performed to analyze the relationship between the expression of CENPF and the prognosis of patients with lung adenocarcinoma. Cox survival hazard ratio was used to analyze the factors affecting the survival of patients. Chi-square analysis was adopted to examine the relationship between CENPF expression and clinicopathological stage and grade of patients. The expression of CENPF in NCI-H2126 cells were knocked out by lentivirus, and then the proliferation, invasion, and migration abilities of the cells were detected. Changes in mRNA expression profiles after CENPF knockout were detected by RNA-seq. Bioinformatics analysis of downstream signaling pathways and the target genes of CENPF was also performed. Western blot was used to verify the target gene. Results CENPF was significantly upregulated in LUAD tumor tissue (P < 0.05) and significantly correlated with pathological stage (P=0.013). The higher expression of CENPF, the worse the prognosis of patients (P=0.01, P=0.027). After the expression was CENPF of knocked out, the cell proliferation, migration, and invasion abilities significantly reduced (P < 0.01). The expression of chemokine pathway genes in cells was enriched significantly (P < 0.001). ACKR3/CXCR7 and CDH2/N-cadherin were significantly downregulated, whereas CDH1/E-cadherin was significantly upregulated. After CENPF was knocked out, ACKR3/CXCR7 and N-cadherin were significantly downregulated, whereas E-cadherin significantly increased. Conclusion The expression of CENPF is negatively correlated with the clinical prognosis of patients with LUAD, and it promotes the occurrence of EMT by regulating the expression levels of N-cadherin and E-cadherin related to EMT through ACKR3/CXCR7.

Chemokine receptor CXCR7 assists CXCR4 in modulating the directional metastasis and EMT of pancreatic cancer / 西安交通大学学报(医学版)

Objective: To investigate the effect of CXCL12/CXCR4/CXCR7 axis on the metastasis and invasion in pancreatic cancer so as to provide new evidence for research on pancreatic cancer metastasis treatment. Methods: MiaPaCa-2 cells were transfected with CXCR4 shRNA and CXCR7 shRNA, and the Transwell assay was used to determine the effects of CXCL12/CXCR4/CXCR7 axis on cell invasion and migration. Quantitative RT-PCR and Western blotting were used to explore the effects of CXCL12/CXCR4/CXCR7 axis on the expressions of invasion-related genes (MMP-2 and uPA) and EMT-related genes (E-cadherin and Vimentin). Results: CXCL12 significantly increased the metastasis and invasion of pancreatic cancer cells. The enhancement of tumor cell invasion was effectively countered by CXCR4 shRNA or CXCR7 shRNA. CXCL12/CXCR4 axis in cancer cells increased the expressions of invasion-related genes (MMP-2 and uPA) and EMT-related genes (E-cadherin and Vimentin). CXCL12/CXCR7 axis only increased the expressions of MMP-2 and uPA. Compared to blocking CXCR4 or CXCR7 alone, the inhibitory effects on invasion-related genes and EMT-related genes were more effective when both CXCR4 and CXCR7 were blocked. Conclusion: CXCL12/CXCR4/CXCR7 axis regulates the EMT, metastasis, and invasion of pancreatic cancer cells.

Effect of SDF-1/CXCR7 on inflammatory cytokine synthesis and secretion in gastric cancer SGC-7901 cells / 西安交通大学学报(医学版)

ABSTRACT:Objective To investigate the effect of SDF-1/CXCR7 on inflammatory cytokine synthesis and secretion in gastric cancer SGC-7901 cells.Methods CXCR7 gene in SGC-7901 cells was silenced by shRNA lentiviral vector and the expression of CXCR7 was detected using Western blot and Real-time PCR.There were four groups as follows:LV-shRNA-NC,LV-shRNA-NC+SDF-1,LV-shRNA-CXCR7,and LV-shRNA-CXCR7+SDF-1 groups.Real-time PCR was used to detect the mRNA expressions of TNF-α,IL-1β,IL-6 and IL-8.ELISA was used to detect the protein levels of TNF-α,IL-1β,IL-6 and IL-8 in the culture supernatant.Western blot was used to detect the protein expressions of NF-κB pathway.Results ① Transfection of SGC-7901 cells with CXCR7-shRNA lentiviral vector resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (all P<0.01).② Compared with those in LV-shRNA-NC group,IL-6 and IL-8 mRNA expressions and protein levels in the culture supernatant were increased in LV-shRNA-NC+SDF-1 group (P<0.01 )and decreased in LV-shRNA-CXCR7 group (P<0.05).Compared with those in LV-shRNA-NC+SDF-1 group,the expressions of IL-6 and IL-8 at mRNA and protein levels in the culture supernatant were significantly cut down in LV-shRNA-CXCR7+SDF-1 group (P<0.01 ).However,the expressions of TNF-αand IL-1βat mRNA and protein levels in the culture supernatant were not significantly changed by SDF-1 and CXCR7 shRNA.③ The protein expressions of nuclear NF-κB p65,t-IκBαand p-IκBαexhibited no significant differences among the four groups.Conclusion SDF-1/CXCR7 can promote the synthesis and secretion of inflammatory cytokines IL-6 and IL-8 in gastric cancer SGC-7 9 0 1 cells through an NF-κB-independent pathway.

Expression of C-X-C chemokine receptor type 7 in otorhinolaryngologic neoplasms

<p><b>INTRODUCTION</b>C-X-C chemokine receptor type 7 (CXCR7) has recently been characterised as a novel receptor for the C-X-C motif chemokine 12 (CXCL12)/stromal cell-derived factor 1-alpha. CXCR7 has been thought to play an important role in the pathogenesis of chronic rhinosinusitis, angiogenesis and tumour metastasis. The present study aimed to examine the expression of CXCR7 in tissue samples of laryngeal cancer and maxillary sinus carcinoma to determine its role in the development of otorhinolaryngologic neoplasms.</p><p><b>METHODS</b>Samples of otorhinolaryngologic neoplasms were obtained from 17 patients with either nasal polyps (n = 7), laryngeal cancer (n = 5) or maxillary sinus carcinoma (n = 5), and who underwent surgical resection at West China Hospital of Sichuan University. Total RNA was isolated and CXCR7 mRNA expression was examined and quantified by relative real-time reverse transcription polymerase chain reaction. A one-way analysis of variance was performed using SPSS Statistics version 11.0 (SPSS Inc, Chicago, IL, USA) to compare the CXCR7 mRNA levels among the three groups of patients.</p><p><b>RESULTS</b>All samples tested positive for CXCR7 mRNA. The quantitative results showed that the CXCR7 mRNA levels were highest in laryngeal cancer and lowest in maxillary sinus carcinoma neoplasms, although there was no significant difference among the three samples.</p><p><b>CONCLUSION</b>CXCL12 and its receptor CXCR7 may contribute to eosinophilic inflammation in patients with chronic sinusitis and nasal polyps. Our results also suggest that CXCR7 may play a role in the progression, metastasis and angiogenesis of otorhinolaryngologic tumours.</p>

Expression and clinical significance of chemokine receptor CXCR7 in epithelial ovarian carcinoma / 第二军医大学学报

Objective To investigate the expression and clinical significance of chemokine receptor CXCR7 in epithelial ovarian carcinoma (EOC) tissues. Methods Immunohistochemical technique was used to detect the protein expression of CXCR7 in 57 EOC tissues, 15 benign ovarian tumor tissues and 15 normal ovarian germinal epithelium tissues. The chi-square test, Kaplan-Meier method and multivariate Cox regression analysis were used to evaluate the relationship between the expression of CXCR7 and clinicopathological features and its prognosis. Results The positive expression rates of CXCR7 in EOC tissue, benign ovarian tumor tissue and normal ovarian germinal epithelium tissue were 68.42% (39/57), 26.67% (4/15) and 13.33% (2/15), respectively (χ2=19.97, P=0.000 1). The positive expression of CXCR7 was significantly correlated with the International Federation of Gynecology and Obstetrics (FIGO) stages, histological grade, lymph node metastasis and peritoneal metastasis (P0.05). Univariate analysis and multivariate analysis results showed that the positive expression of CXCR7, FIGO stage, histological grade, lymph node metastasis and peritoneal metastasis were statistically correlated with the overall survival and disease-free survival of EOC patients. Kaplan-Meier curve revealed that both overall survival and disease-free survival in EOC patients with positive expression of CXCR7 were significantly lower than those in EOC patients with negative CXCR7 (P<0.05). ConclusionThe high expression of CXCR7 is closely related to the recurrence and metastasis in EOC patients, which indicates that CXCR7 might be a potential predictor and an important therapeutic indicator of EOC.

Expression and functional roles of the chemokine receptor CXCR7 in acute myeloid leukemia cells

BACKGROUND: The C-X-C chemokine receptor 7 (CXCR7) has been shown to be a decoy receptor for CXCR4 in certain cell types. We investigated the expression status and functional roles of CXCR7 in acute myeloid leukemia (AML) cells in vitro. METHODS: CXCR7 mRNA was knocked down in AML cells by using small interfering RNA (siRNA) technology, and subsequent biological alterations in the cells were evaluated in vitro. RESULTS: All AML cell lines examined in this study (U937, K562, KG1a, HL-60, and MO7e) and primary CD34+ cells obtained from patients with AML expressed CXCR7 mRNA at various levels. Western blotting showed that all AML cells produced CXCR7. Furthermore, all AML cells expressed CXCR7 in both the cytoplasm and on the cell surface at various levels. Stromal cell-derived factor-1 (SDF-1; C-X-C motif ligand 12 (CXCL12)) induced internalization of cell surface CXCR7. However, neither hypoxia nor the examined hematopoietic growth factors (interleukin-1beta (IL-1beta), IL-3, IL-6, granulocyte-colony-stimulating factor, granulocyte, macrophage-colony-stimulating factor, and stem cell factor) and proinflammatory cytokines (interferon-gamma, transforming growth factor-beta, and tumor necrosis factor-alpha) were found to alter cell surface CXCR7 expression. The transfection of AML cells with CXCR4 siRNA, but not CXCR7 siRNA, significantly impaired the CXCL12-induced transmigration of the cells. The transfection of AML cells with CXCR7 siRNA did not affect the survival or proliferation of these cells. Knockdown of CXCR7, but not CXCR4, induced the upregulation of CXCL12 mRNA expression and CXCL12 production in AML cells. CONCLUSION: CXCR7 is involved in the regulation of autocrine CXCL12 in AML cells.

Effects of RNA interference targeting the chemokine receptor 7 gene on the invasion and migration of the human melanoma cell line M14 / 中华皮肤科杂志

Objective To explore the effects of targeted silencing of the chemokine receptor 7 (CXCR7)gene on the invasion and migration of the melanoma cell line M14. Methods Western-blot analysis was performed to determine the protein expression of CXCR7 in melanoma cell lines M14 and A375, and CXCR7-overexpressing M14 cells were used in this study. Cultured M14 cells were divided into three groups: experimental group transfected with a small interfering RNA(siRNA)targeting CXCR7(CXCR7-siRNA), negative control group transfected with a negative control siRNA, blank control group receiving no treatment. Real-time quantitative PCR and Western-blot analysis were conducted to determine the mRNA and protein expressions of CXCR7 respectively in M14 cells, Transwell chambers were used to evaluate the invasive activity of M14 cells, and wound healing assay to estimate the migratory activity of M14 cells. Results The experimental group showed significantly decreased mRNA and protein expressions of CXCR7 compared with the negative control group and blank control group (CXCR7 mRNA: 0.412 ± 0.023 vs. 1.211 ± 0.117 and 1.000 ± 0.102, F = 30.068, P = 0.001; CXCR7 protein: 0.144 ± 0.005 vs. 1 and 1.016 ± 0.004, F =11 485.5, P = 0.000). The number of M14 cells crossing the polycarbonate membrane per high-power field (× 200)was significantly smaller in the experimental group than in the negative control group and blank control group (20.617 ± 1.503 vs. 42.000 ± 6.018 and 43.627 ± 2.152, F = 32.416, P = 0.001). Similarly, the number of migrating M14 cells in wound healing assay was significantly decreased in the experimental group compared with the negative control group and blank control group (15.00 ± 1.10 vs. 44.90 ± 2.20 and 45.30 ± 2.30, F = 2 411.945, P = 0.000). Conclusion Targeted silencing of the CXCR7 gene can significantly inhibit the invasion and migration of M14 cells in vitro, which may provide a potential target for the treatment of cutaneous melanoma.

Roles of the CXCL12-CXCR4/CXCR7 chemokine axis in colorectal carcinoma / 医学研究生学报

Colorectal carcinoma ( CRC ) is one of the malignant tumors with the highest morbidity and mortality rates in the world.The invasion and metastasis of CRC are the major reasons for treatment failure and death .It was believed that CXCR 4 was the exclusive receptor for CXCL12.However, recent studies have identified CXCR7 as a second receptor for CXCL12.Both chemokine ax-es of CXCL12/CXCR4 and CXCL12/CXCR7 closely correlate with the proliferation , migration, adhesion of tumor cells and the forma-tion of tumor-associated vessels in CRC , which may become new significant targets for anti-cancer and anti-metastatic treatment of CRC.

Expression and function of CXCR2, CXCR7 of acute leukemic cells in rat

OBJECTIVE@#To investigate the expression and function of chemokine receptor CXCR2 and CXCR7 in the rat with acute leukemia.@*METHODS@#Flow cytometry and RT-PCR were used to detect the CXCR2, CXCR7 expression on the bone marrow cell surface of the acute leukemia group and the control group.@*RESULTS@#The bone marrow cell surface CXCR2, CXCR7 relative fluorescence intensity of the observation group was significantly higher than the control group (P<0.05). The CXCR7 expression of the extramedullary infiltration group was significantly higher than non-extramedullary infiltration group (P<0.05). The CXCR2, CXCR7mRNA median expression level of the observation group was higher than the control group. The CXCR2 expression and CXCR7 expression of the observation group was positively correlated, and the correlation coefficient was 0.782 (P<0.01).@*CONCLUSIONS@#The chemokine receptor CXCR2 and CXCR7 are highly expressed in acute leukemia, which may be associated with the occurrence of leukemia.

Speciifc targeted-intervention effect of CXCR 7-shRNA mediated by low viral carrier plus Rhizoma Paridis total saponin on tumor cells:an experimental study / 中国生化药物杂志

Objective To investigate the CXCR 7 protein expression when CXCR 7-shRNA transfected into human gastric cancer cell which mediated with lentivirus vector combined with Rhizoma Paridis Total Saponin. Methods Three shRNA sequences of CXCR 7 and one negative control sequence were designed and synthesized, and recombinant lentiviral vectors with pSilencerTM 4.1 system were established. Transfection of HEK 293 T cells and packaging viral were finished and the titers were detected. Transfection of all recombinant lentiviral vectors and negative control vector were finished and expression of CXCR 7 mRNA were detected by RT-PCR method. Silence efficiency in groups were determined and the expression vector with highest silence efficiency was selected for next experiments. To detect the effect of SGC 7901 cell proliferation by CXCR 7-shRNA transfection and combined with Rhizoma Paridis Total Saponin intervention with MTT. To detect the effect of SGC 7901 cell expression of protein by CXCR 7-shRNA transfection and combined with Rhizoma Paridis Total Saponin intervention with Western blot. Results The packaging of three lentiviral vector and negative control sequence are successful which is confirmed by gene sequencing and the titer are 4.9×108 pfu/mL, 3.6×108 pfu/mL, 5.2×108 pfu/mL, 2.0×108 pfu/mL respective. The expression quantity of CXCR 7 mRNA in positive groups are lower than negative control group(P<0.05)and inhibition ratio to CXCR 7 in CXCR 7-shRNA-1 and combined with Rhizoma Paridis Total Saponin intervention group is higher than the other two groups(P<0.05). The proliferation level of tumour cell is significant reduction after CXCR 7-shRNA-1 transfection and have a significant difference comparing to the group without transfection(P<0.05). The expression of CXCR 7 protein is significant reduction after CXCR 7-shRNA-1 transfection comparing to the group without virus vector and negative control group and have a significant difference(P<0.05). Conclusion The construction of three CXCR 7-shRNA lentiviral expression vector are successful and expression level of protein and CXCR 7 mRNA are down-regulated effectively after transfection and combined with Rhizoma Paridis Total Saponin intervention. It maybe means that CXCR 7 gene takes an important role in the process of gastric cancer proliferation and invasion.This is foundation for further study of gastric cancer gene therapy using CXCR 7/CXCL 12 biological axis as a target.

Effect of stromal cell derived factor-1α/CXCR4/CXCR7 axis on migration of the bone marrow mesenchymal stem cells / 解剖学报

Objective To investigate expression of CXCR4 and CXCR7 protein and mRNA, which are the receptors of stromal cell derived factor-1α(SDF-1α), in the bone marrow mesenchymal stem cells (BMSCs);to explore the role of SDF-1α/CXCR4/CXCR7 axis in migration of BMSCs in vitro and the possible mechanism .Method BMSCs were isolated from rats and cultured in vitro.CD29, CD44 and CD34 of the cells were identified by flow cytometry .CXCR4-selective antagonist AMD 3100 and CXCR7-specific neutralizing antibody were applied to block CXCR 4 and CXCR7 respectively.The expressions of CXCR4 and CXCR7 mRNA and protein on BMSCs were detected with RT-PCR and Western blotting .Transwells chamber test was used to observe the migration of BMSCs .The BMSCs were divided into the BMSCs group ( A ) , the AMD3100 pretreated BMSCs group ( B ) , the CXCR7-specific neutralizing antibody pretreated BMSCs group(C), the AMD3100 +CXCR7-specific neutralizing antibody pretreated BMSCs group ( D).Result Flow cytometry showed that the expressions of CD 44 and CD29 were positive, while the expression of CD34 was negative in the third passage of BMSCs (P3-BMSCs).CXCR4 and CXCR7 protein and mRNA were both expressed in P3-BMSCs. Compared with the A group, the expression of CXCR4 and CXCR7 protein declined significantly in the B group and the D group;the protein expression of CXCR7 in the C group was lower compared with the A group (P<0.05).However, the expression of CXCR4 mRNA and CXCR7 mRNA had no significant difference between groups .SDF-1αfactor promoted migration of BMSCs ( P <0.05 ).Compared with the 0μg/L group, the numbers of migrated cells were increased significantly in both of the 10μg/L group and the 100μg/L group ( P<0.01 ) .The number of migration of BMSCs was significantly higher in the 100μg/L group than that of the 10μg/L group ( P <0.01 ) .AMD3100 and CXCR7-specific neutralizing antibody both inhibited significantly the migration of BMSCs ( P<0.05 ) , and the attenuate effect was more significant when they worked together ( P<0.05 ) .Conclusion CXCR4 and CXCR7 receptors are co-expressed in P3-BMSCs;the SDF-1αfactor can promote the migration of BMSCs in the concentration dependent manner ;SDF-1α/CXCR4/CXCR7 axis is involved in the migration of BMSCs , and both of the CXCR4 and CXCR7 receptors have a synergistic promoting effect to the BMSCs migration .

Chemokine receptor CXCR7 in tumor invasion and metastasis / 国际肿瘤学杂志

Chemokines are major regulators of cell transformation and adhesion.Recent study has demonstrated that CXCR7 can bind to CXCL11 and CXCL12 with high affinity,and the activated CXCR7 may influence tumor invasion and metastasis by regulating extracellular matrix (ECM) and epithelial-mesenchymal transition (EMT) and other signal transduction pathways.Therefore,in-depth study of the molecular mechanisms of CXCR7 in tumor invasion and metastasis may provide a more effective theoretical basis for tumor treatment.

Chemokine receptor CXCR7 and tumor / 国际肿瘤学杂志

CXCR7 is a new receptor of chemokine CXCL12 after CXCR4.The present study shows that CXCL12/CXCR7 biological axis has important influence to the development of a variety of tumors,similar to CXCL12/CXCR4 biological axis.CXCR7 is present in many kinds of tumor tissues and tumor cells widely,and plays an important role in tumor cells growth,proliferation,adhesion and migration.Restraining CXCR7 expression or blocking the CXCR7 signaling pathways may offer new strategies for the treatment of tumors.

Expression of CXCR7 protein in human bladder cancer and its clinical significance / 中华泌尿外科杂志

Objective To explore the expression of CXCR7 in bladder cancer and analyze its clinical significance and relationship with bladder cancer recurrence. Methods The expressions of CXCR7 protein in 148 specimens of bladder cancer and 30 specimens of normal bladder tissues were detected by immunohistochemical staining and its clinical significance was then analyzed. Results The expression of CXCR7 protein was higher in bladder cancers than in the adjacent normal tissues (P＜0.01). CXCR7 protein expression rates were 49. 4% and 71.2% in mutifocal tumors and unifocal tumors, while 34.0%, 65.8% and 78. 6% in G1, G2, and G3 tumors, respectively (P＜0. 01). Expression of CXCR7 protein was higher in muscle invasive bladder cancers than in non-muscle invasive bladder cancers (72. 7% versus 51.9% ,P＜0.05). In patients followed up for 2-95 months, CXCR7 protein expression was significantly higher in patients with recurrence than with non-recurrence (64. 1% versus 32.5%, P＜0.01). Kaplan-Meier analysis and the log-rark test showed that the recurrence-free survival was significantly different between the group of lower CXCR7 expression group and the higher expression group (P＜0.01). Conclusions The expression of CXCR7 protein is high in bladder cancer and the analysis of CXCR7 protein expression is potentially valuable in prognostic evaluation of bladder cancers. CXCR7 may play a role in the development of bladder urothelial cell cancer.

The chemokine receptors CXCR4 and CXCR7 in cancer / 中国癌症杂志

CXCR4 has long been considered as the unique receptor of CXCL12,and CXCL12/CXCR4 axis plays a prominent role in tumorigenesis.However,a novel receptor for CXCL12,named CXCR7,has been recently identified and also plays an important role in tumorigenesis.This review summarized current studies regarding the functions of CXCR4 and CXCR7 in cancer and the recent therapeutic approaches that target these receptors or their ligands.