Recent studies revealed that tumor-associated macrophages (TAMs) play a decisive role in the regulation of tumor progression by manipulating tumor oncogenesis, angiogenesis and immune functions within tumor microenvironments. Signal transducer and activator of transcription 3 (STAT3), which is a point of convergence for numerous oncogenic signalling pathways, is constitutively activated both intumor cells andin immune cells in the tumormicroenvironment. TAMs serve as the major components of niche microenvironments regulating cancerstemcell functions. Activation of the Stat3in TAM caused to cancerstemcell-specific fashion trigger tumorigenesis and anticancer drug resistance intumor. The main aim of this study is evaluation of stat3 gene expression intumormicroenvironment macrophages.
The above findings indicate that Oct1 controls multiple markers and activities associated with stem cells, but do not address whether Oct1 controls the stemcell phenotype itself. We previously showed that stable Oct1 RNAi in luciferase-expressing A549 cells reduces tumorigenicity in xenograft assays without affecting growth rates in culture . In these experiments, 2610 6 cells were transplanted andtumor mass was partially reduced (by approximately 60%). We hypothesized that differences intumor initiating frequency underlie this effect. We injected reduced numbers of cells harboring scrambled or Oct1-specific shRNAs into opposite flanks of nude mice. Cells were pre-treated with doxycycline for 48 hr and injected into immunocompromised mice maintained on doxycycline. Using 1610 6 cells, both the scrambled and Oct1-specific shRNA-expressing cells engrafted 15/15 recipient mice. A further tenfold reduction resulted in 12/ 13 mice engrafted using cells expressing scrambled shRNAs while cells expressing Oct1-specific shRNAs engrafted 4/13 mice in the contralateral flank (Figure 5A, 5B). The remaining six mice showed no evidence of engraftment as assessed by visual inspection, palpation or bioluminescence. 50,000 cells engrafted poorly (4/15) using scrambled shRNAs and not at all with Oct1- specific shRNAs. Using even fewer cells, 0/15 mice engrafted regardless of Oct1 status (Figure 5B). These findings allowed us to calculate that Oct1 shRNA reduces the frequency of initiating cells from ,1/96,000 to ,1/350,000 (Figure 5A). We also over- expressed Oct1 in luciferase-expressing A549 cells. In this case no pre-treatment took place and the mice were not administered doxycycline. Oct1 overexpression was moderate (Figure 3F). This level of Oct1 expression leads to a .2-fold increase in TIC frequency (Figure 5C). Similar results were obtained with MDA- MB-231 human breast adenocarcinoma cells and mammary fat pad engraftment. Using Oct1 lentiviral knockdown, TIC frequen- cy shifted downwards from ,1/50,000 to ,1/175,000 (Figure 5D–5F). Oct1 overexpression increased TIC frequency ,3-fold from ,1/50,000 to ,1/17,000 (Figure 5G–5H).
Inflammation has been known as one of the main keys to the establishment and pro- gression of cancers. Chronic low-grade inflammation is also a strategic condition that underlies the causes and development of metabolic syndrome and obesity. Moreover, obesity has been largely related to poor prognosis of tumors by modulating tumormicroenvironment with secretion of several inflammatory mediators by tumor-associated adipocytes (TAAs), which can modulate and recruit tumor-associated macrophages. Thus, the understanding of cellular and molecular mechanisms that underlay and link inflammation, obesity, andcancer is crucial to identify potential targets that interfere with this important route. Knowledge about the exact role of each component of the tumormicroenvironment is not yet fully understood, but the new insights in literature highlight the essential role of adipocytes and macrophages interplay as keyfactor to determine the fate of cancer progression. In this review article, we focus on the functions of adipocytes and macrophages orchestrating cellular and molecular mechanisms that lead to inflammatory modulation intumormicroenvironment, which will be crucial to cancer establishment. We also emphasized the mechanisms by which the tumor pro- motes itself by recruiting and polarizing macrophages, discussing the role of adipocytes in this process. In addition, we discuss here the newest possible anticancer therapeutic treatments aiming to retard the development of the tumor based on what is known about cancer, adipocyte, and macrophage polarization.
Proteomic profiling of the estrogen/tamoxifen-sensitive MCF-7 cell line and its partially sensitive (MCF-7/LCC1) and fully resistant (MCF-7/LCC9) variants was performed to identify modifiers of endocrine sensitivity in breast cancer. Analysis of the expression of 120 paired phosphorylated and non-phosphorylated epitopes inkey oncogenic andtumor suppressor pathways revealed that STAT1 and several phosphorylated epitopes (phospho-STAT1(Tyr701) and phospho-STAT3(Ser727)) were differentially expressed between endocrine resistant and parental controls, confirmed by qRT-PCR and western blotting. The STAT1 inhibitor EGCG was a more effective inhibitor of the endocrine resistant MCF-7/LCC1 and MCF-7/LCC9 lines than parental MCF-7 cells, while STAT3 inhibitors Stattic and WP1066 were equally effective in endocrine-resistant and parental lines. The effects of the STAT inhibitors were additive, rather than synergistic, when tested in combination with tamoxifen in vitro. Expression of STAT1 andSTAT3 were measured by quantitative immunofluorescence in invasive breast cancers and matched lymph nodes. When lymph node expression was compared to its paired primary breast cancer expression, there was greater expression of cytoplasmic STAT1 (,3.1 fold), phospho-STAT3(Ser727) (,1.8 fold), and STAT5 (,1.5 fold) and nuclear phospho-STAT3(Ser727) (,1.5 fold) in the nodes. Expression levels of STAT1 andSTAT3 transcript were analysed in 550 breast cancers from publicly available gene expression datasets (GSE2990, GSE12093, GSE6532). When treatment with tamoxifen was considered, STAT1 gene expression was nearly predictive of distant metastasis-free survival (DMFS, log-rank p = 0.067), while STAT3 gene expression was predictive of DMFS (log-rank p,0.0001). Analysis of STAT1 andSTAT3 protein expression in a series of 546 breast cancers also indicated that high expression of STAT3 protein was associated with improved survival (DMFS, p = 0.006). These results suggest that STAT signaling is important in endocrine resistance, and that STAT inhibitors may represent potential therapies in breast cancer, even in the resistant setting.
regulator element in breast carcinoma formation in a Her2-transgenic mouse mammary tumor model. (70) Moreover, downregulation of GnT-V in mouse mammary cancercell lines showed a significant suppression of tumor growth and metastasis. (69) The progression of breast cancerand its metastasis induced by a viral oncogene in transgenic mice is considerably noticeably suppressed in MGAT5-deficient background. (71) Also, GnT-V-mediated glycosylation has been shown to regulate the cancerstemcell compartment andtumor progression trough WNT signaling. (72) As opposed to the function of GnT-V, GnT-III (which is encoded by MGAT3) catalyses the addition of bisecting GlcNAc N-glycans in a β-1,4 linkage, inhibiting the additional processing and elongation of N-glycans, like the β1,6-branching structures. GnT-III assumes an opposite role to GnT-V incancer, as it is involved in the suppression of cancer metastasis. (73) It was tested in mouse melanoma cells the transfection of MGAT3 into this high metastatic potential tissue, and it resulted in a significant reduction of β1,6GlcNAc branching (as a result of the enzymatic competition between GnT-III and GnT-V), which led to a notable suppression of lung metastasis in mice. The mechanism of tumor metastasis suppression carried out by GnT-III is through the regulation of key proteins, such as EGFR, integrins and cadherins (63,74), as will be explained further.
crine and autocrine communications play an important role, increasing the invasion and metastasis potential intumor cells [58– 60]. Interestingly, substantial production of tumor promoting factor (MMP-13) and marked levels of NF-kB were further pronounced in the presence of 5-FU. Moreover, many different tumor cells activate constitutive inducible anti-apoptotic NF-kB for their survival [43,61]. This result seems to be paradox at first, as 5-FU would be expected to impede tumor promotion. However, taken into consideration that 5-FU also stimulated enrichment of CSC marker positive cells in high density tumormicroenvironment co-cultures, these results might be regarded as a defense reaction/response of the tumorcell culture and the tumormicroenvironment. In fact, it has been reported that in contrast to the differentiated population of tumor cells, cancerstem cells are characterized by increased resistance to cytotoxic chemotherapeutic agents , enhanced ability to form colono- spheres [6,13,63] and induce remission, providing basis for why cancer cells cannot be completely destroyed by conventional chemotherapeutic agents [57,64]. Indeed, we could further show that combinational treatment of curcumin and 5-FU dramatically suppressed tumor promoting factors, activation of NF-kB signaling pathway and enrichment of CSCs in high density tumormicroenvironment co-cultures. These results demonstrate once more the potent chemosensitizing effect of curcumin on CRC, and to our knowledge show for the first time the modulating effect of curcumin on the crosstalk between the CRC/CSCs and the fibroblasts in the tumormicroenvironment, creating an adequate climate for more effective chemotherapeutic action of 5-FU to specifically target chemoresistant colorectal stem cells.
The BT20 breast cancer cells used in these experiments were isolated from a breast tumor that was triple negative for the expression of Her2, estrogen receptor and progesterone recep- tor. Such triple negative tumors often express EGFR . Triple negative breast cancer is also associated with increased stromal reaction with inflammatory component and a poor progno- sis. In our co-culture model, BT20 cells secreted increased amounts of various inflammatory cytokines, including the pro-inflammatory cytokine IL6 (S4 Fig). IL6, has been reported to in- duce cell proliferation in various cancers . We also observed the activation of STAT3, a transcription factor that is downstream of the IL6 receptor, in the co-cultured BT20 cells, indi- cating the activation of this pathway (S5C Fig). Blocking IL6 resulted in a significant decrease in the survival of BT20 cells that were co-cultured with fibroblasts, indicating that the activa- tion of STAT3and the subsequent increase incell proliferation are mediated by IL6. Impor- tantly, the BT20 co-cultures also secreted EGF but did not respond to Erbitux treatment. In this context, it is important to note that the activation of the IL6 pathway has been implicated in the resistance to Trastuzumab (anti-Her2) in PTEN-deficient tumor cells . Thus, IL6 se- cretion in BT20 co-cultures may be responsible for the resistance to Erbitux in our model. These data suggest that tumor cells can acquire resistance to therapeutic agents by utilizing al- ternative pathways depending on the availability of the corresponding ligands and the compo- sition of the TME.
The capability of reconstituting a heterogenous mass is a definition of cancerstem cells, and we found that isolated ALDH+ prostate cancer cells were capable of self-renewal and re- establishment of the parental cell line and possessed high tumorigenicity in vivo andin vitro. The ALDH+ population of LNCaP-IL6 cells had a high expression of the putative prostate cancerstem cells markers CD44 and integrin a2b1  compared to the ALDH2 population. However, we did not detect CD133 expression in the ALDH+ or ALDH2 prostate cancer cells. This is in agreement with the recent study by Pfeiffer and Schalken  suggesting that CD133 is not a marker for stem cells in prostate cancercell lines. Increased JAK/STAT3and NF-kB activity is expressed instem cells andtumor initiating stem-like cells in prostate cancer [18,36] and our findings of active STAT3and NF- kB in ALDH+ cells is in accordance with this suggestion. Taken together, our results, and others , indicate that ALDH expression can be a means of identifying cancerstemcell-like cells in prostate cancer.
activates transcription of the target genes of proliferation, anti-apoptosis, or cell cycle progression by binding directly to high-affinity DNA binding sites or by associating with other transcription factors. In addition to its diverse biologi- cal functions involved incell proliferation, differentiation, apoptosis, inflammation, and oncogenesis, accumulating evidence suggests that STAT3 also plays an important role in angiogenesis in both physiological and pathologi- cal situations (7-13). Several studies have reported that STAT3 directly regulates vascular endothelial growth factor A (VEGFA) expression to induce tumor angiogenesis in breast cancer, gastric cancerand hepatocellular carci- noma (4,5,14). In pancreatic cancer, STAT3-targeting RNA interference (RNAi) inhibits angiogenesis by suppressing the expression of matrix metalloproteinase-2 (MMP2) (15). In human melanoma brain metastases, activated STAT3 contributes to tumor angiogenesis by regulating the expression of basic fibroblast growth factor (FGF2) (3). These results indicate that STAT3 signaling interferes with the process of angiogenesis in various cancers through different mechanisms.
Os fatores de transcrição Ets são considerados alvos difíceis porque possuem pobre atividade enzimática, a regulação de seus genes alvo é complexa e existe nas ligações uma ampla rede de proteínas parceiras e necessárias para as suas funções (1). No entanto, encontramos correlação entre a média geométrica da expressão relativa dos fatores de transcrição ETs de PDEF e ETV4 entre os tecidos benigno e maligno no CaP (p<0,001 e p<0,001, respectivamente). E a expressão relativa da média geométrica do PDEF foi 102 vezes maior no tecido maligno prostático em relação ao benigno, enquanto que no ETV4 foi 6,09 vezes maior no tecido maligno prostático em relação ao benigno. Muitos artigos relatam que a expressão de PDEF é diminuída a medida que o tumor se torna mais agressivo e invasivo, se correlacionando com estadiamento e grau tumoral, como no câncer de próstata, mama, ovário e cólon (261), (279), (283), (285), (288). Porém o inverso também é demonstrado em câncer de próstata, mama e ovário, sendo sugerido até uma atividade supressora de tumor para PDEF (261), (285), (326-329). O coeficiente de correlação entre os dois fatores de transcrição no tecido benigno prostático foi muito forte com p<0,001, e o coeficiente de correlação no tecido maligno prostático foi forte com p=0,009; isso comprova que ambos são fatores de transcrição multifuncionais e que podem atuar em vias que se correlacionam, como as das metaloproteinases que leva a migração, invasão celular e metástases.
Brother of the regulator of imprinting sites (BORIS) also designed as CTCFL, CCCTC-binding factor-like, is a DNA- binding protein with functions incancer not fully understood. CTCF is a highly conserved, ubiquitously expressed, multifunc- tional chromatin factor that plays a role as a tumor suppressor gene [1–3]. BORIS is a mammalian paralog of CTCF, they share the same 11 zinc-finger domain but differ at N- and C- termini. Within this zinc-finger domain, BORIS and CTCF exhibit 70% of homology . In normal tissues, BORIS expression is restricted to germ cells, where it is involved in epigenetic reprogramming [4,5]. BORIS is expressed in spermatocytes during male germ line development, apparently in absence of CTCF . In tumors, BORIS is aberrantly expressed and its transcription was detected at different levels in several cancercell lines andin primary tumors . Due to its restricted expression in normal germinal tissues and its re-expression in a wide variety of tumors, BORIS belongs to cancer testis antigen (CTA) family. It has been shown that BORIS induced expression of other CTA genes, as MAGE-A1, NY-ESO- 1 [7,8] and SPANX  but not in all tumors [10,11]. In addition, we previously showed that BORIS activated hTERT expression by
various messages to promote or to inhibit antitumor responses. Despite plethora of investigations on various EVs, it seems we still need to fully decode the main messages of these silently whispering vesicles and examine their potentials in diagnosis and treatment of diseases (in particular malignancies) in which the involved cells use such intricate bio-machineries for their communications. The conducted studies together with the growing body of evidence indicate that EXOs provide great potentials as a novel nanoscale cellular machineries for various diagnostic and therapeutic purposes. However, there exist some striking questions to be addressed. What if these EVs convey signals to suppress the immunosurveillance or danger signals to make TME much more permissive? Is it likely that they shuttle danger signals to neighboring cells/tissue? Do the malignant cells use such capacity for the migration and hence colonization into the neighboring cells/tissue? What are the main roles of lipid rafts , membranous caveolae and clathrin coated-pitsd in this process? Taken all, in the best scenario, we may capitalize on these cell-free vaccination system. And, if we exploit these paramount and worth pursuing nanoshuttles for cancer immunotherapy and vaccination, which issues need to be considered to improve the exosomal immunogenicity? To the best of our knowledge, these EVs need to be optimized in terms of (a) the antigen-loading efficiency, (b) the compositions, morphology and sizes, (c) the in vivo trafficking, and finally (d) the biological fates and impacts in the target cells.
Uma vez que a angiogênese é um importante evento na progressão do adenocarcinoma pancreático, há uma aparente contradição com a sua característica de hipovascularização na tomografia computadorizada com contraste endovenoso. Wang et al. (2003) ressaltaram que a atenuação do tumor pancreático é o resultado da combinação da DMV intra-tumoral, da desmoplasia e do tecido pancreático normal peritumoral que inclui, também, os vasos de maior calibre intra e peritumorais. Além disso, a perfusão do tecido tumoral depende da organização da drenagem venosa e linfática (linfangiogênese) que é significativamente desorganizada nas neoplasias (Jain et al., 1988; Cuenod et al., 2006; Kadota et al., 2008).
Prostate-specific membrane antigen (PSMA) is the most extensively studied biomarker and antigen of prostate cancer. It is overexpressed in almost all prostate cancers, and the expression level increases with prostate cancer progression. PSMA is also highly expressed in the neovasculature of solid tumours including prostate cancer. As a result, numerous PSMA-specific ligands have been discovered for prostate cancer diagnosis and therapy, and one of them has been approved for clinical use. Moreover, a number of other PSMA-specific ligands are currently evaluated in clinical studies. In this review we discuss four major types of PSMA-specific ligands, including antibody, aptamer, peptide, and small molecule inhibitor. Their emerging applications in prostate cancer diagnosis, targeted drug delivery, and therapy are also discussed.
However, our study has several limitations, as we could not prevent all potential bias among these studies. Although 21 studies were included in the final analysis, the average sample size was small (mean,98). For articles without HR and 95%CI, we contacted the corresponding author, but received no reply. Consequently, we extrapolated these metrics from the data or curves in these articles indirectly, which could be another potential source of bias. Different methods of examining bFGF expression, primary antibody source and concentration, and threshold cut-off values may introduce more bias as well. IHC is a relatively complicated tech- nique with many steps and is observer-dependent. Moreover, our meta-analysis data did not include information on age, smoking status, tumor size and other factors, which might result in confounding bias. Lastly, we included studies which dichotomized bFGF expression into the high and low expression group. Other studies that included bFGF expression as a continuous variable were excluded because the data was not extractable.
The addition of plerixafor shows a consistent increase in collection rate success, reducing the number of apheresis and not increasing toxicity. Strategies of preemptive use of plerix- afor have been considered a promising way to optimize and rationalize the use of this agent in patients who have high chance of failure with classic mobilization based on G-CSF with or without chemotherapy. This strategy would reduce failure in mobilization, especially in poor mobilizers, ensuring collection and transplantation as well as reducing time and costs of the mobilization procedure. However, external valid- ity of these algorithms is limited, so it is recommended that each institution sets up a strategy appropriate to its standards for the preemptive use of plerixafor.
Studies show the important roles for some chemokines in hematopoiesis and organ development. Hematopoiesis is an active progression controlled by a variety of cytokines. It is well clear that the chemokine family plays an critical role in this regulatory system. At least 25 chemokines of the CC, CXC and C subgroups have been found to sup- press the in vitro proliferation of myeloid progenitor cells (12, 13). However, the in vivo evidence based on knock- out mice studies provides data of contrasting hematopoi- etic effect for only few of the chemokines and their recep- tors. For example, CCL3 (MIP-1α) arrests cell cycling and reduces the bone marrow progenitor cells number in mice (14). Mice depleted of CCR1, as CCL3 receptor, display enhanced lineage-committed myeloproliferation and leukocyte mobilization into the blood stream (15). CXCL12 (SDF-1) is constitutively expressed by stromal cells in bone marrow, promotes proliferation of B cell progenitors (16), and recruit hematopoietic precursors to the bone marrow in embryogenesis time (17). Mice de- ficient in CXCL12 (SDF-1) or its receptor (CXCR4) die before birth with in B lymphopoiesis and myelopoiesis deficiency and also with an incomplete development of the cardiac septums and cerebellum development, it show the involvement of CXCL12 /CXCR4 pair in a number of vital developmental processes (18-20). The thymus is organ for T cell development and it expresses mRNA for several chemokines with lymphocyte-attracting proper- ties, that well described (21,22). Some chemokines con- trol the development and organization of secondary lym- phoid organs such as peripheral lymph nodes and Peyer’s patches (23,24).
Surgical resection remains the most effective treatment for patients with early-stage NSCLC. However, the reported 5- year survival rates of patients with stage I disease are between 60% and 90% (26,27). The rate of recurrence in patients with stage I NSCLC suggests that systemic tumorcell dissemination occurs before tumor resection but remains undetectable by current clinical techniques (28). Based on the high frequency of recurrence in patients with stage I NSCLC, which is presumed to be a localized disease, several clinical trials have been conducted to analyze the benefit of adjuvant chemotherapy for stage I NSCLC patients who have undergone complete resection, although the results are conflicting. A retrospective analysis per- formed by the Cancerand Leukemia Group B (CALGB) 9633, which randomly assigned patients with stage IB NSCLC to receive adjuvant chemotherapy or no additional treatment, demonstrated a possible benefit for patients with tumors larger than 4 cm but not for patients with smaller primary tumors (29). Meta-analysis of the Lung Adjuvant Cisplatin Evaluation (LACE) suggested a poorer outcome for patients with stage IA tumors who received adjuvant chemotherapy when compared with patients with stage IA tumors who did not receive treatment (30). Thus, the current standard of care for stage I NSCLC patients who have undergone resection remains post-operative surveillance regardless of the possibility of occult ‘‘micrometastatic’’ disease. If appropriate methods were available to determine which stage I NSCLC patients would benefit from post- operative chemotherapy, then patients with an increased risk for recurrence could be selected for therapy and would be expected to experience substantial clinical benefit from adjuvant systemic chemotherapy. Therefore, methods to
As a new emerging therapy target, the FGFR gene has drawn much interest for developing specific inhibitors such as the multiple target inhibitors dovitinib, Ki23057, and ponatinib, and the highly selective inhibitors AZD4547 and BGJ398. Several preclinical studies have shown the striking therapeutic efficacy of AZD4547 and BGJ398 on FGFR gene–amplified cancers both in vitro andin vivo [15,36,37]. Some ongoing clinical trials have been summarized in a published paper . Recently, a phase II study was designed to assess the activity of the FGFR inhibitor AZD4547 in patients with FGFR1- and FGFR2-amplified breast, squamous lung, or stomach cancers, whose cancers had progressed following previous chemotherapy (NCT01795768). Our data indicated that both FGFR1 and FGFR2 amplification were associated with poor survival in breast, lung, and gastric cancers. It is therefore reasonable to conduct more clinical trials that set FGFR copy number as an inclusion criterion. More importantly, our data highlighted the need for collaborative efforts in addressing FGFR as a therapeutic target. For example, the sample size for clinical trial evaluating anti-FGFR2 drug efficacy is about
The expression levels of ERBB3 incancer tissues compared to matched normal tissues were highly variable, ranging from 0.06 to 60.2-fold change respectively (Fig 1). 38.8% (19/49) of tumours had increased ERBB3 expression levels (>2-fold) compared to matched normal tis- sues (Fig 1). The median level of ERBB3 expression in adenocarcinomas (0.141) was signifi- cantly higher than that in normal tissues (0.079) (Wilcoxon matched-pairs test, p<0.0005) (Fig 1). Similar results were obtained in adenomas compared to normal tissues, although the differ- ence did not reach statistical significance (Paired Student’s T test, p = 0.061) probably due to the low number of adenoma samples in this patient cohort (n = 4) (Fig 2A). The mean value of the logarithm of the fold change (0.587) was significantly different from 0 (p = 0.009, one-sam- ple t-test). Linear regression modelling indicated that ERBB3 expression levels were also signif- icantly higher in stage IV tumours compared to stage I tumours (Coefficient = 1.36, (95% confidence interval (CI) 0.07, 2.64), p = 0.039, data not shown). There were no other significant relationships between ERBB3 expression levels and clinical markers such as disease stage, grade and TNM stages. Increased expression of ERBB3 in adenomas and adenocarcinomas compared to normal tissues was confirmed by immunohistochemistry (Fig 2B). ERBB3 was predominantly localised to the apical and basolateral cell surface of normal intestinal epithelial cells and was detected at the cell surface in the majority of tumours with some diffuse cyto- plasmic staining apparent in a few cases (S1 Fig).