The CC chemokine RANTES in breast carcinoma progression: regulation of expression and potential mechanisms of promalignant activity

Breast cancer progression may be affected by various cellular components expressed by the tumor cells and/or by microenvironmental factors. Many studies report the correlation between breast cancer progression and monocyte infiltration into the tumor site. We have identified recently the CC chemokine regulated on activation, normal T cell expressed and secreted (RANTES), a major monocyte chemoattractant expressed by breast tumor cells, as a potential contributor to breast cancer progression. In the present study, analysis of the regulation of RANTES expression demonstrates that the expression of RANTES in breast tumor cells is elevated significantly and in a synergistic manner by IFN-gamma and tumor necrosis factor-alpha. Identification of the mechanisms by which RANTES may contribute to breast cancer progression included the analysis of the potential ability of RANTES to act in paracrine and indirect mechanisms, as well as directly on the tumor cells, to promote disease progression. Our results suggest that breast tumor cell-derived RANTES may promote breast cancer progression by its partial contribution to monocyte migration into breast tumor sites. Moreover, RANTES promotes the expression of matrix metalloproteinase (MMP) 9 by THP-1 monocytic cells and elevates vascularity in chick chorioallantoic membrane assays. Tumor necrosis factor-alpha, a major monocyte-derived cytokine, was found to promote the expression of MMP9 and MMP2 by MCF-7 and T47D breast adenocarcinoma cells, respectively, and to induce the de novo expression of an additional proteolytic enzyme by T47D cells, presumably MMP9. The possibility that RANTES may act directly on breast tumor cells was supported by detection of the expression of the CCR5 RANTES receptor in biopsy sections of breast cancer patients and by the ability of RANTES to promote the expression of MMP9 by MCF-7 cells. In all, our study suggests that the expression of RANTES by breast tumor cells results not only in monocyte migration to the tumor site but also in protumorigenic activities of RANTES and of proinflammatory cytokines that may facilitate metastasis formation and contribute to disease progression.     

趋化因子CCL8与肿瘤发生发展的研究进展

趋化因子CCL8是趋化因子CC家族中的一员,可以在炎症反应和肿瘤免疫等方面发挥重要作用。近来许多研究数据表明,CCL8与人类肿瘤的发生发展密切相关,它在肿瘤微环境中扮演了不可或缺的角色,一方面CCL8可以促进肿瘤细胞的浸润转移,另一方面又可以抑制肿瘤的发生发展。本文主要阐述了目前研究发现的CCL8在肿瘤中的作用,包括肿瘤的发生发展、浸润转移等,以上研究不仅可以帮助我们更加深入的了解肿瘤形成的机制,而且对于寻找更加有效的肿瘤新疗法有着重要的意义。     

CXCL16/CXCR6 chemokine signaling mediates breast cancer progression by pERK1/2-dependent mechanisms

Our previous studies demonstrate that CXCL6/CXCR6 chemokine axis induces prostate cancer progression by the AKT/mTOR signaling pathway; however, its role and mechanisms underlying invasiveness and metastasis of breast cancer are yet to be elucidated. In this investigation, CXCR6 protein expression was examined using high-density tissue microarrays and immunohistochemistry. Expression of CXCR6 shows a higher epithelial staining in breast cancer nest site and metastatic lymph node than the normal breast tissue, suggesting that CXCR6 may be involved in breast cancer (BC) development. In vitro and in vivo experiments indicate that overexpression of CXCR6 in BC cells has a marked effect on increasing cell migration, invasion and metastasis. In contrast, reduction of CXCR6 expression by shRNAs in these cells greatly reduce its invasion and metastasis ability. Mechanistic analyses show that CXCL16/CXCR6 chemokine axis is capable of modulating activation of RhoA through activating ERK1/2 signaling pathway, which then inhibits the activity of cofilin, thereby enhancing the stability of F-actin, responsible for invasiveness and metastasis of BC.Taken together, our data shows for the first time that the CXCR6 / ERK1/2/ RhoA / cofilin /F-actin pathway plays a central role in the development of BC. Targeting the signaling pathway may prove beneficial to prevent metastasis and provide a more effective therapeutic strategy for BC.     

The chemokine CCL5 as a potential prognostic factor predicting disease progression in stage II breast cancer patients.

PURPOSE: The aim of this study was to determine the prognostic value of the chemokine CCL5, considered as a promalignancy factor in breast cancer, in predicting breast cancer progression and to evaluate its ability to strengthen the prognostic significance of other biomarkers. EXPERIMENTAL DESIGN: The expression of CCL5, alone and in conjunction with estrogen receptor (ER)-alpha, ER-beta, progesterone receptor (PR), and HER-2/neu (ErbB2), was determined in breast tumor cells by immunohistochemistry. The study included 142 breast cancer patients, including individuals in whom disease has progressed. RESULTS: Using Cox proportional hazard models, univariate analysis suggested that, in stage I breast cancer patients, CCL5 was not a significant predictor of disease progression. In contrast, in stage II patients, the expression of CCL5 (CCL5(+)), the absence of ER-alpha (ER-alpha(-)), and the lack of PR expression (PR(-)) increased significantly the risk for disease progression (P = 0.0045, 0.0041, and 0.0107, respectively). The prognostic strength of CCL5, as well as of ER-alpha(-), improved by combining them together (CCL5(+)/ER-alpha(-): P = 0.0001), being highly evident in the stage IIA subgroup [CCL5(+)/ER-alpha(-) (P = 0.0003); ER-alpha(-) (P = 0.0315)]. In the stage II group as a whole, the combinations of CCL5(-)/ER-alpha(+) and CCL5(-)/PR(+) were highly correlated with an improved prognosis. Multivariate analysis indicated that, in stage II patients, ER-alpha and CCL5 were independent predictors of disease progression. CONCLUSIONS: CCL5 could be considered as a biomarker for disease progression in stage II breast cancer patients, with the CCL5(+)/ER-alpha(-) combination providing improved prediction of disease progression, primarily in the stage IIA subgroup.     

CC chemokine ligand 7 expression in liver metastasis of colorectal cancer

The main cause of death for colorectal cancer (CRC) patients is the development of metastatic lesions at sites distant from the primary tumor. Therefore, it is important to find biomarkers that are related to the metastasis and to study the possible mechanisms. Recent data have shown that soluble attractant molecules called chemokines support the metastasis of certain cancers to certain organs. To identify molecular regulators that are differentially expressed in liver metastasis of CRC, PCR array analysis was performed and CC chemokine ligand 7 (CCL7) showed remarkable overexpression in liver metastatic tumor tissues. To validate the results of the PCR array, 30 patients with primary CRC and liver metastases were selected. Immunohistochemistry and real-time PCR analysis showed that CCL7 was expressed in normal colonic epithelium and the expression was higher in liver metastases compared to primary CRC (p<0.001). Real-time PCR showed that the expression of CCR1, CCR2 and CCR3 was also higher in liver metastases compared to primary CRC (p=0.001, p=0.033 and p<0.001, respectively). In conclusion, correlation of CCL7 overexpression and its receptor expression with colon cancer liver metastasis suggests that CCL7 as a novel target in liver metastasis of CRC may be of potential clinical value for the prevention of hepatic recurrences.     

Analysis of CC chemokine and chemokine receptor expression in solid ovarian tumours

To understand the chemokine network in a tissue, both chemokine and chemokine receptor expression should be studied. Human epithelial ovarian tumours express a range of chemokines but little is known about the expression and localisation of chemokine receptors. With the aim of understanding chemokine action in this cancer, we investigated receptors for CC-chemokines and their ligands in 25 biopsies of human ovarian cancer. CC-chemokine receptor mRNA was generally absent from solid tumours, the exception being CCR1 which was detected in samples from 75% of patients. CCR1 mRNA localised to macrophages and lymphocytes and there was a correlation between numbers of CD8(+) and CCR1 expressing cells (P = 0.031). mRNA for 6 CC-chemokines was expressed in a majority of tumour samples. In a monocytic cell line in vitro, we found that CCR1 mRNA expression was increased 5-fold by hypoxia. We suggest that the CC-chemokine network in ovarian cancer is controlled at the level of CC-chemokine receptors and this may account for the phenotypes of infiltrating cells found in these tumours. The leukocyte infiltrate may contribute to tumour growth and spread by providing growth survival factors and matrix metalloproteases. Thus, CCR1 may be a novel therapeutic target in ovarian cancer.     

Protein expression of the chemokine, CCL28, in human colorectal cancer

Accumulating evidence points out that chemokines play important roles in colorectal mucosal immunity by recruiting leukocytes into and out of the lamina propria adjacent to the epithelium. The chemokine, CCL28, which is expressed by epithelial cells within colorectal mucosa is thought to have dual roles as a chemoattractant for leukocytes expressing CCR10 and/or CCR3 and a mediator of antimicrobial activity. To date, there is little known about the presence of CCL28 in colorectal cancer. The objective of this study was to determine whether the CCL28 protein level is altered in colorectal tumours (n=76) compared with paired normal mucosa. Further, the plasma CCL28 levels from patients with colon and rectal cancer were also examined. Immunohistochemistry revealed heterogeneous CCL28 protein expression in tumour and normal epithelial cells. Analyses by ELISA showed that the CCL28 protein level in colon tumours was significantly (P<0.001) lower than in normal tissue and that the difference in CCL28 protein level between rectal tumour and normal tissue was not significant. Patients with a tumour localized in the colon had significantly (P<0.05) higher plasma CCL28 protein levels than patients with a tumour localized in the rectum. These discrepancies may suggest that there are different mechanisms and signalling pathways involved in the pathogenesis of cancer in the colon and rectum and thereby differentially promote the expression and secretion of CCL28 protein. Further studies on CCL28 protein in colorectal cancer with an extended number of patients are necessary to conclude whether tissue levels and plasma concentrations carry significant clinical relevance.     

Clinical utility of circulating matrix metalloproteinase-7 (MMP-7), CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer

Ovarian cancer remains a highly lethal disease. The aim of the present study was to evaluate the usefulness of measuring serum matrix metalloproteinase-7 (MMP-7), CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) in comparison with serum cancer antigen 125 (CA 125) for diagnosis of epithelial ovarian cancer (EOC). This study included 51 patients with EOC, 27 patients with benign ovarian lesions and 29 healthy volunteers. Serum CA 125 was determined by microparticle enzyme immunoassay, while serum MMP-7, CCL18 and CCL11 were measured using enzyme-linked immunosorbent assay. The sensitivity and specificity were 86.3% and 92.9% for CA 125, 80.4% and 87.5% for MMP-7, 84.3% and 91.1% for CCL18 and, 68.6% and 62.5% for CCL11. Combination of CA 125, MMP-7, CCL18 and CCL11 gave a promising sensitivity of 100%, but specificity was decreased to 60.7%. The combined use of serum CA 125, MMP-7, CCL18 and CCL11 effectively detected early stages EOC with high sensitivity of 94.4%. Our data indicate that serum MMP-7, CCL18 and CCL11, in combination with CA 125 could be useful in diagnosis of EOC.     

结直肠癌趋化因子MCP-1的表达及其与巨噬细胞浸润的关系

Objective： To study the expression of MCP-1 in colorectal carcinoma and its relationship to the infiltration of the macrophage and to the biological behaviour of infiltration and metastasis of colorectal carcinoma. Methods： The expression of the MCP-1 mRNA was assessed in colorectal carcinoma collected freshly from surgical specimen by RT-PCR and the expression of the MCP-1 protein was assessed in colorectal carcinoma collected from surgical specimen by immunohistochemistry. The tumor infiltrating cell and macrophage were also investigated by immunohistochemistry. Results： All the 12 specimens of colorectal carcinoma detected by RT-PCR expressed the MCP-1 mRNA; MCP-1 protein was detected in 90% （36/40） cases of the tumor; The expression of the MCP-1 protein in colorectal carcinoma correlated negatively with its state of metastasis and the Dukes＇ stage. But a postive correlation was found between the expression of MCP-1 and the infiltrated macrophage. The stronger expression of MCP-1, the more number of the infiltrated macrophage. Conclusion： The expression of chemokine MCP-1 in colorectal carcinoma may influence its biological behaviour of infiltration and metastasis, and can attract the immuno-cell to the local of the tumor, such as Macrophage.     

miR-221/222 induces pancreatic cancer progression through the regulation of matrix metalloproteinases

MicroRNAs are involved in the initiation and progression of pancreatic cancer. In this study, we showed that miR-221/222 is overexpressed in pancreatic cancer. MiR-221/222 overexpression significantly promoted pancreatic cancer cell proliferation and invasion while inhibiting apoptosis. The expression of the matrix metalloproteinases (MMPs) MMP-2 and MMP-9 was increased in miR-221/222 mimic-transfected pancreatic cancer cells. Validation experiments identified TIMP-2 as a direct target of miR-221/222. These data indicate that overexpressed miR-221/222 may play an oncogenic role in pancreatic cancer by inducing the expression of MMP-2 and MMP-9, thus leading to cancer cell invasion.     

抑制CC类趋化因子配体5基因表达对乳腺癌细胞增殖能力的影响

目的 探讨抑制CC类趋化因子配体5(CCL5)基因表达对人乳腺癌细胞增殖能力的影响.方法 用特异性CCL5 RNA干扰(RNAi)序列慢病毒载体感染人乳腺癌细胞MCF-7和MDA-MB-231,分别为KD1组和KD2组;另在MCF-7和MDA-MB-231细胞中分设阴性病毒载体感染的阴性对照组(NC1组和NC2组)和未感染组(CON1组和CON2组).采用实时定量逆转录聚合酶链反应(RT-PCR)检测转染病毒后乳腺癌细胞中CCL5的表达,四甲基偶氮唑蓝(MTT)比色法和流式细胞术(FACS)分析细胞的增殖情况,平板克隆形成实验观察细胞的克隆形成能力.结果 CCL5 RNAi慢病毒可显著降低MCF-7和MDA-MB-231细胞中CCL5基因的表达.MTF法检测结果显示,在不同的培养时间,MCF-7和MDA-MB-231细胞的KD组、NC组与CON组细胞培养上清A值差异均无统计学意义(均P＞0.05).FACS分析结果显示,KD1组、NC1组和CON1组的增殖指数(PI)值分别为0.48±0.03、0.43±0.01和0.45±0.02;KD2组、NC2组和CON2组的PI值分别为0.48±0.02、0.44±0.05和0.47±0.02(两两比较,均P＞0.05).荧光显微镜下观察显示,KD组的克隆体积及每克隆的细胞数明显小于NC组和CON组.KD1组和KD2组克降数目(0.34±0.08和0.33±0.10)明显少于NC1组(0.81±0.12)、NC2组(0.97±0.09)、CON1组(0.92±0.12)和CON2组(1.04±0.07),差异有统计学意义(P＜0.05).结论 CCL5基因的表达下调对乳腺癌MCF-7和MDA-MB-231细胞群体倍增时间无明显影响,但可显著降低细胞的克隆形成能力,从而使肿瘤细胞的恶性增殖受到抑制.

Abstract：

Objective To investigate the effect of suppression of CCI5 ligand gene on the proliferation of human breast cancer cells. Methods A lentiviral vector carrying a short interfering RNA (siRNA) targeting CCL5 was transfected into human breast cancer cell line MCF-7 and MDA-MB-231 cells.The expression of CCL5 mRNA in the cells was detected by real-time PCR. The proliferation of MCF-7 and MDA-MB-231 cells was assessed by MTT assay and FACS assay, and the colony formation ability of both cell lines were measured, respectively. Results Real time PCR showed a good knockdown effect of CCL5 in both cell-lines. Colony-forming assay showed that the ability of colony formation of MCF-7/CCL5-siRNA and MDA-MB-231/CCL5-siRNA was decreased markedly. The colony number of MCF-7/CCL5-siRNA group was (0. 34 ± 0. 08), significantly lower than 0. 81 ± 0. 12 in the MCF-7/CCL5-N group and 0.92 ± 0.12 in the MCF-7 group (P ＜ 0. 05). The colony number of MDA-MB-231/CCL5-siRNA group was 0. 33 ± 0. 10,significantly lower than 0.97 ±0.09 in the MDA-MB-231/CCL5-N group and 1.04 ±0.07 in the MDA-MB-231 group (P ＜0.05). However, MTT assay revealed that the proliferation of MCF-7/CCL5-siRNA cells was not significantly different from that of MCF-7/CCL5-N or MCF-7 cells, respectively (P ＞0.05), and the same result was found in MDA-MB-231 cells. FACS assay showed that the proliferation index (PI) of groups MCF-7/CCL5-siRNA, MCF-7/CCL5-N and MCF-7 were 0.48 ± 0. 03, 0. 43 ± 0. 01 and 0.45 ±0. 02. The PI of groups MDA-MB-231/CCL5-siRNA, MDA-MB-231/CCL5-N and MDA-MB-231 cells were 0. 48 ± 0.02, 0.44 ± 0.05 and 0. 47 ± 0. 02. There was no statistical difference among them ( P ＞ 0.05 ).Conclusion The down-regulation of CCL5 gene in human breast cancer cells may significantly suppress their colony formation ability, rather than affecting their population doubling time to some extent.     

CXCL14 expression and potential function in pancreatic cancer

LAPTM4B was proven to overexpress in hepatocellular carcinoma and relate to differentiation. We immunohistochemically investigated the expression and potential clinicopathological and prognostic significance of LAPTM4B encoded protein, LAPTM4B-35, in extrahepatic cholangiocarcinoma (EHCC) for specimens from consecutive 81 patients. LAPTM4B-35 staining was positive in cancer tissues from 59 patients (72.8%), including 12 with score 1, 22 with score 2 and 25 with score 3. No positive staining was found in non-cancer epithelia. The staining score in cancer tissues was not only significantly associated with TNM staging, histological grade, perineural and lymph node invasion (P < 0.05), but also of comprehensive prognostic implications, including integrated estimation with CA19-9. These data established that LAPTM4B-35 positively expressed in a great portion of EHCC and might be a novel molecular maker of progression, invasiveness and poor prognosis.     

ENO1, a potential prognostic head and neck cancer marker,promotes transformation partly via chemokine CCL20 induction

The success of using glycolytic inhibitors for cancer treatment depends on studying the individual role of frequently deregulated glycolytic genes in cancer. This report aims to study the prognostic implication, and determine the cellular role and action mechanism of glycolytic ENO1 overexpression in head and neck cancer. The relationship of ENO1 mRNA expression in 44-pair clinical specimens with patient clinicopathologic characteristics was analysed by semi-quantitative RT-PCR, Kaplan–Meier survival curve and Cox model analyses. Following ectopic ENO1 expression or knockdown, we studied the proliferative, migratory, invasive, colony-forming and tumourigenic abilities of ENO1-genetically altered cells. DNA microarray analysis was used to identify downstream targets responsible for the ENO1 action in the cells. The expression of ENO1 mRNA was increased in 68% of tumour (T) specimens when compared to their normal (N) counterparts, and positively associated with clinical progression (p < 0.05). High ENO1 expression (T/N ? 2) was frequently observed in the patients with large primary tumours, late clinical stages or advanced neck metastasis. Moreover, high ENO1 patients had significantly poorer clinical outcomes than low expressers (T/N < 2). Ectopic ENO1 expression stimulated cell transformation, invasion and tongue tumour formation. ENO1 knockdown abrogated the stimulation. Suppression of ENO1-induced proinflammatory CCL20 chemokine expression significantly attenuated its stimulatory effects on cell transformation and invasion. A concordant expression of ENO1 and CCL20 was validated both in ENO1-expresing cells and in clinical specimens. Together, we demonstrate a prognostic role of ENO1 overexpression in head and neck cancer and ENO1-mediated promotion of cell transformation and invasion partly via induced CCL20 expression.     

The inflammatory chemokines CCL2 and CCL5 in breast cancer

A causal role was recently attributed to inflammation in many malignant diseases, including breast cancer. The different inflammatory mediators that are involved in this disease include cells, cytokines and chemokines. Of these, many studies have addressed the involvement and roles of the inflammatory chemokines CCL2 (MCP-1) and CCL5 (RANTES) in breast malignancy. While minimally expressed by normal breast epithelial duct cells, both chemokines are highly expressed by breast tumor cells at primary tumor sites, indicating that CCL2 and CCL5 expression is acquired in the course of malignant transformation, and suggesting that the two chemokines play a role in breast cancer development and/or progression. Supporting this possibility are findings showing significant associations between CCL2 and CCL5 and more advanced disease course and progression. Furthermore, studies in animal model systems have shown active and causative roles for the two chemokines in this disease. In line with the tumor-promoting roles of CCL2 and CCL5 in breast cancer, the two chemokines were shown to mediate many types of tumor-promoting cross-talks between the tumor cells and cells of the tumor microenvironment: (1) they shift the balance at the tumor site between different leukocyte cell types by increasing the presence of deleterious tumor-associated macrophages (TAM) and inhibiting potential anti-tumor T cell activities; (2) of the two chemokines, mainly CCL2 promotes angiogenesis; (3) CCL2 and CCL5 which are expressed by cells of the tumor microenvironment osteoblasts and mesenchymal stem cells play a role in breast metastatic processes. In addition, both chemokines act directly on the tumor cells to promote their pro-malignancy phenotype, by increasing their migratory and invasion-related properties. Together, the overall current information suggests that CCL2 and CCL5 are inflammatory mediators with pro-malignancy activities in breast cancer, and that they should be considered as potential therapeutic targets for the limitation of this disease.     

BCL2 expression correlates with metastatic potential in pancreatic cancer cell lines

BACKGROUND: Programmed cell death (termed apoptosis) regulates normal tissue homeostasis. Loss of local paracrine signals and intercellular adhesion molecules are potent inducers of apoptosis and thereby eliminate normal cells that may have escaped beyond the confines of the local organ environment. Dysregulation in the expression of the BCL2 gene family, the prototypic regulators of apoptosis, is a common occurrence in cancer and imparts resistance to standard triggers of apoptosis. Therefore, the authors sought to examine whether abnormal BCL2 gene family expression correlated with resistance to apoptosis and increased metastatic potential in pancreatic carcinoma. METHODS: The authors examined BCL2 expression and apoptotic sensitivity in three panels of human pancreatic cancer cell lines that possess varying metastatic potential. Stable transfectants were generated that overexpress BCL2. These transfectants were then analyzed for differences in metastasis formation in athymic mice. RESULTS: Among the isogenic panels of pancreatic cancer cell lines, BCL2 expression levels correlated with metastatic potential. Highly metastatic variants of each family of cell lines were more resistant to induction of apoptosis. Finally, using the BCL2 transfectant in a xenograft model, elevated BCL2 expression led to a higher incidence of metastases. CONCLUSIONS: The authors conclude that increased BCL2 expression correlates with apoptotic resistance and metastatic potential; dysregulation of BCL2 expression may be involved in the metastatic progression of pancreatic carcinoma.     

分化型甲状腺癌患者停用甲状腺素后短期甲减对血脂的影响

Objective  

The aim of this study was to analyze the expression of CC chemokine receptor 7 (CCR7) in pulmonary tumor tissue and metastasized lymph nodes of non-small cell lung cancer (NSCLC), explore the relationship between the expressions of CCR7 in pulmonary tumor tissue and metastasized lymph nodes, and discuss the significance.