VEGFR3 suppression through miR-1236 inhibits proliferation and induces apoptosis in ovarian cancer via ERK1/2 and AKT signaling pathways.

Vascular endothelial growth factor receptor 3 (VEGFR3) is expressed in cancer cell lines and exerts a critical role in cancer progression. However, the signaling pathways of VEGFR3 in ovarian cancer cell proliferation remain unclear. This study aimed to demonstrate the signaling pathways of VEGFR3 through the upregulated expression of miR-1236 in ovarian cancer cells. We found that the messenger RNA and protein of VEGFR3 were expressed in the ovarian cancer cell lines, but downregulated after microRNA-1236 (miR-1236) transfection. The inhibition of VEGFR3, using miR-1236, significantly reduced cell proliferation, clonogenic survival, migration, and invasion ability in SKOV3 and OVCAR3 cells (p < 0.01). The flow cytometry results indicated that the rate of apoptotic cells in SKOV3 (38.65%) and OVCAR3 (41.95%) cells increased following VEGFR3 inhibition. Moreover, VEGFR3 stimulation (using a specific ligand, VEGF-CS) significantly increased extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (AKT) phosphorylation (p < 0.01), whereas VEGFR3 suppression reduced p-ERK1/2 (67.94% in SKOV3 and 93.52% in OVCAR3) and p-AKT (59.56% in SKOV3 and 78.73% in OVCAR3) compared to the VEGF-CS treated group. This finding demonstrated that miR-1236 may act as an endogenous regulator of ERK1/2 and AKT signaling by blocking the upstream regulator of VEGFR3. Overall, we demonstrated the important role of the miR-1236/VEGFR3 axis in ovarian cancer cell proliferation by regulating the ERK1/2 and AKT signaling that might be an effective strategy against ovarian cancer.

Circulating plasma miR222-3P status and its potential diagnostic performance in prostate cancer.

BACKGROUND: Although studies suggest that miR222-3p is dysregulated in prostate cancer (PC) cells and tissues, the possible changes in the level of miR222-3p in the plasma samples of PC patients remained unclear. The present study aimed to evaluate the diagnostic value of the plasma miR222-3p expression level as a potential biomarker in PC, benign prostatic hyperplasia (BPH) and healthy people. METHODS: Blood samples were collected from 100 adult males (54 patients with PC, 27 patients with BPH and 19 healthy individuals) referred to our affiliated hospital. The expression level of miR222-3p was evaluated using a quantitative reverse transcription-polymerase chain reaction. Receiver operating characteristic curves were used to evaluate miR222-3p diagnostic accuracy for discriminating between the PC, BPH and healthy individuals. RESULTS: The expression level of miR222-3p was significantly higher in PC patients compared to healthy individuals as a fold change of 5.3 (p = 0.009), but not for BPH individuals. The diagnostic value of the plasma miR222-3p for discrimination of the PC patients from healthy individuals was reasonable [cut-off value (fold change relative to miR16-5p) = 1.69, area under the curve = 0.73, sensitivity = 0.75 and specificity = 0.74]. CONCLUSIONS: Circulating plasma miR-222-3p significantly upregulated in PC patients, but not in BPH ones. Besides these preliminary results showed that miR222-3p has the potential to discriminate PC patients from healthy ones. Addittional studies with a larger sample size are required to confirm these data.

Body composition and serum levels of matrix metalloproteinase-9, adiponectin and AMP-activated protein kinase in breast cancer survivors.

Background: Available data suggest that obesity is related to changes in the several adipocyte-derived proteins levels, which are involved in cancer recurrence. The purpose of this work was to investigate the correlation between obesity with metalloproteinase-9 (MMP-9), adiponectin and adiponectin and AMP-activated protein kinase (AMPK) levels by comparing serum levels of MMP-9, AMPK in normal weight and obese breast cancer survivors. Materials and Methods: In this cross-sectional study, 30 normal weight breast cancer survivors (body mass index [BMI] 18.5-25 kg/m2) and 30 obese breast cancer survivors (BMI ≥30 kg/m2) were investigated. Anthropometric parameters and serum levels of MMP-9, adiponectin, and AMPK were compared between the two groups. Results: No differences were detected in the serum levels of MMP-9, adiponectin, and AMPK in obese patients and normal weight patients (P > 0.05). There were no correlations between MMP-9, adiponectin, and AMPK levels with anthropometric measurements in two groups (P > 0.05). Conclusion: We found that there was a lack of correlation between obesity measures and serum levels of MMP-9, adiponectin, and AMPK. In breast cancer survivors, it seems that circulating levels of adiponectin, AMPK, and MMP-9 do not change in obesity state.

Circulating status of microRNAs 660-5p and 210-3p in breast cancer patients.

BACKGROUND: MicroRNAs (miRs), which are stable in the blood, comprise small non-coding RNAs that regulate gene expression. They have important roles in almost all biological pathways, especially in cancer-relevant processes, and have an abnormal expression in breast cancer. In recent studies, the aberrant expression level of various microRNAs has been demonstrated in human cancer. In the present study, the status of serum microRNA-210-3p and microRNA-660-5p expression levels in breast cancer patients was determined compared to healthy controls. METHODS: Serum samples were collected from 40 newly diagnosed breast cancer patients and 40 healthy controls. A real-time quantitative polymerase chain reaction was utilized to detect the expression levels of these microRNAs. Data analysis was conducted with p < 0.05 being considered statistically significant. RESULTS: The data obtained showed that serum levels of miR-660-5p and miR-210-3p were significantly up-regulated in breast cancer patients compared to healthy controls (p < 0.001 and p = 0.001, respectively). In addition, significant up-regulation was observed in the early stage (in situ, stage I and II) of breast cancer patients (n = 25) compared to healthy (n = 40) controls (p < 0.001 and p < 0.05, respectively). Receiver-operating characteristic curve analysis indicated that the serum miR-660-5p and miR-210-3p levels have reasonable sensitivity (79% and 68%) and specificity (61% and 51%) for the detection of breast cancer patients (area under the receiver-operating curve = 0.774 and 0.716, respectively). CONCLUSIONS: Although the results show a reasonable diagnostic accuracy of these microRNAs for detection of breast cancer in this small and preliminary study, further large-scale studies are essential to confirm the presented results.

Evaluation of the plasma level of long non-coding RNA PCAT1 in prostatic hyperplasia and newly diagnosed prostate cancer patients.

BACKGROUND: Prostate cancer (PCa) is generally detected by prostate-specific antigen (PSA) as one of the most widely applied tumor markers over decades for its high sensitivity. Nevertheless, it causes overtreatment or an unnecessary biopsy because of its limited specificity. PCa-associated ncRNA transcript 1 (PCAT1), the newly identified long non-coding RNA (lncRNA) has been reported to associate with the progress of PCa. In vitro studies proposed that PCAT-1 may be an appealing candidate for diagnostic accuracy improvement with regard to its notable overexpression in PCa cells. The present study aimed to evaluate the diagnostic potential of the plasma PCAT1 expression levels in PCa patients in comparison to benign prostatic hyperplasia (BPH) patients and healthy controls. METHODS: The plasma lncRNA PCAT1 level was measured by a real-time quantitative reverse transcriptase-polymerase chain reaction in 40 men newly diagnosed with PCa, 20 patients with BPH and 20 healthy subjects. The results were analyzed statistically using SPSS, version 25 (IBM Corp., Armonk, NY, USA). RESULTS: The expression of PCAT1 was significantly higher in healthy subjects compared to BPH patients (p = 0.03). The diagnostic accuracy of the plasma lncRNA PCAT-1 for discrimination of the healthy subjects than BPH patients was reasonable (area under the receiver operating characteristic curve = 0.799; sensitivity = 71%; specificity = 74%; negative predictive value = 74%; positive predictive value = 71%). CONCLUSIONS: It appears that the plasma levels of PCAT1 expression have reasonable diagnostic accuracy for the discrimination of healthy individuals compared to those with BPH, although no significant difference of PCAT1 expression levels was observed in comparisons between the PCa with BPH and normal groups.

CRISPR/Cas9 technology as a potent molecular tool for gene therapy.

Clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR-Cas9) is an RNA-guided gene editing tool which offers several advantageous characteristics in comparison with the conventional methods (e.g., zinc finger nucleases and transcription activator-like effector nucleases) such as cost-effectiveness, flexibility, and being easy-to-use. Despite some limitations such as efficient delivery and safety, CRISPR-Cas9 is still the most convenient tool for gene editing purposes. Due to the potential capability of the CRISPR-Cas9 system in genome editing and correction of casual mutations, it can be considered as a possible therapeutic system in the treatment of disorders associated with the genome mutations and in particular cancer treatment. In this review, we will discuss CRISPR-Cas-based gene editing along with its classifications and mechanism of action. Furthermore, the therapeutic application of the CRISPR-Cas9 system in mutational disorders, delivery systems, as well as its advantages and limitations with a special emphasis on cancer treatment will be discussed.

RAS/MAPK signaling functions in oxidative stress, DNA damage response and cancer progression.

Mitogen-activated protein kinase (MAPK) signaling pathways organize a great constitution network that regulates several physiological processes, like cell growth, differentiation, and apoptotic cell death. Due to the crucial importance of this signaling pathway, dysregulation of the MAPK signaling cascades is involved in the pathogenesis of various human cancer types. Oxidative stress and DNA damage are two important factors which in common lead to carcinogenesis through dysregulation of this signaling pathway. Reactive oxygen species (ROS) are a common subproduct of oxidative energy metabolism and are considered to be a significant physiological modulator of several intracellular signaling pathways including the MAPK pathway. Studies demonstrated that the MAP kinases extracellular signal-regulated kinase (ERK) 1/2 and p38 were activated in response to oxidative stress. In addition, DNA damage is a partly common circumstance in cell life and may result in mutation, cancer, and even cell death. Recently, accumulating evidence illustrated that the MEK/ERK pathway is associated with the suitable performance of cellular DNA damage response (DDR), the main pathway of tumor suppression. During DDR, the MEK/ERK pathway is regularly activated, which contributes to the appropriate activation of DDR checkpoints to inhibit cell division. Therefore, the aim of this review is to comprehensively discuss the critical function of MAPK signaling in oxidative stress, DNA damage, and cancer progression.

Crosstalk between Phosphoinositide 3-kinase/Akt signaling pathway with DNA damage response and oxidative stress in cancer.

The phosphatidylinositol 3-kinases (PI3K)/Akt signaling pathway is one of the well-characterized and most important signaling pathways activated in response to DNA damage. This review discusses the most recent discoveries on the involvement of PI3K/Akt signaling pathway in cancer development, as well as stimulation of some important signaling networks involved in the maintenance of cellular homeostasis upon DNA damage, with an exploration of how PI3K/Akt signaling pathway contributes to the regulation of modulators and effectors underlying DNA damage response, the intricate, protein-based signal transduction network, which decides between cell cycle arrest, DNA repair, and apoptosis, the elimination of irreparably damaged cells to maintain homeostasis. The review continues by looking at the interplay between cell cycle checkpoints, checking the repair of damage inflicted to the DNA before entering DNA replication to facilitate DNA synthesis, and PI3K/Akt signaling pathway. We then investigate the challenges the cells overcome to ameliorate damages induced by oxidative activities, for example, the recruitment of many pathways and factors to maintain integrity and hemostasis. Finally, the review provides a discussion of how cells use the PI3K/Akt signaling pathway to regulate the balance between these networks.

The roles of FGF21 in atherosclerosis pathogenesis.

FGF21 is a peptide hormone that regulates homeostasis of lipid and glucose as well as energy metabolism. It is mainly expressed and secreted in liver and adipose tissues, and it is expressed in lower amounts in the aorta. Recent clinical and preclinical studies indicate increased serum FGF21 levels in atherosclerosis patients. Also, FGF21 therapy has been reported to reduce the initiation and progression of atherosclerosis in animal models and in vitro studies. Moreover, growing evidence indicates that administration of exogenous FGF21 induces anti-atherosclerotic effects, because of its ability to reduce lipid profile, alleviation of oxidative stress, inflammation, and apoptosis. Therefore, FGF21 can not only be considered as a biomarker for predicting atherosclerosis, but also induce protective effects against atherosclerosis. Besides, serum levels of FGF21 increase in various diseases including in diabetes mellitus, hypertension, and obesity, which may be related to initiating and exacerbating atherosclerosis. On the other hand, FGF21 therapy significantly improves lipid profiles, and reduces vascular inflammation and oxidative stress in atherosclerosis related diseases. Therefore, further prospective studies are needed to clarify whether FGF21 can be used as a prognostic biomarker to identify individuals at future risk of atherosclerosis in these atherosclerosis-associated diseases. In this review, we will discuss the possible mechanism by which FGF21 protects against atherosclerosis.

Multiple Functions of Long Non-Coding RNAs in Oxidative Stress, DNA Damage Response and Cancer Progression.

In addition to aberrant alternation of transcriptome, it is now suggested that dysregulation of the non-coding transcripts, particularly long non-coding RNAs (lncRNAs), which comprise the majority of the genome, is contributed to cancer initiation and progression. As the result of recent huge efforts, the possible roles of numerous lncRNAs in the human cancers were characterized, as well as various strategies with inhibitory effects to target these transcripts on the transformed cells. Moreover, DNA damage response (DDR) pathway is a complex regulatory network responsible for the identification of disruptions in DNA structure, integrity and stability- it is reported to be associated with the up-regulation and down-regulation of lncRNAs. This review explores the involvement of the various lncRNAs in different human cancers, afterwards discusses the association of the lncRNAs expression with the DDR and oxidative stress, which are implicated in a myriad pathophysiological and physiological intra- and extracellular damages. J. Cell. Biochem. 119: 223-236, 2018. © 2017 Wiley Periodicals, Inc.

ABO Blood Group and Prevalence of Osteoporosis and Osteopenia in the Elderly Population: An Amirkola Health and Ageing Project (AHAP)-Based Study.

Osteoporosis is known as a degenerative disease of the skeletal system and its main complication is fracture, which influences quality of life in the elderly. There are 4 major blood groups in humans based on the presence of A and B antigens. According to the investigations, there are reported relations between blood types and some diseases. In this study, the association between the ABO blood group and the prevalence of osteoporosis and osteopenia in an elderly population was investigated. Medical records of 990 elderly people were investigated in a cross-sectional study and the association between their blood group and the incidence of osteoporosis and osteopenia was analyzed using SPSS version 17.0 (SPSS Inc., Chicago, IL, USA). The results showed that ABO blood groups had no association with the prevalence of osteoporosis in both elderly men and women. The association between age and osteoporosis was significant and the association between this disorder and gender was significant too. The results also indicate that there is no association between RH+ and RH- blood types and osteoporosis and osteopenia in both men and women. Based on this finding, it would be reasonable to conduct extensive studies.

Evaluation of scalp hair nickel and chromium level changes in patients with fixed orthodontic appliance: a one-year follow-up study.

BACKGROUND AND OBJECTIVES: The release of metal ions from orthodontic appliances is part of the dissolution and biomechanical processes of alloys. Nickel (Ni) and chromium (Cr) are the elements commonly used in the manufacture of various components of fixed orthodontic appliances, including bands, brackets and wires. This study was aimed to measure the Ni and Cr ions levels in the scalp hair of patients treated with fixed orthodontic appliances in comparison of the control group. MATERIALS AND METHODS: The patient group consisted of 24 patients treated with fixed orthodontic appliances for one year, while the control group included 28 healthy individuals without orthodontic appliances. Analysis of the Cr and Ni was performed using atomic absorption spectrophotometer by graphite furnace method. The data were analyzed via student and paired samples t-test and ANOVA repeated measurement test. RESULTS: After one year, the levels of Ni and Cr in two groups showed significant differences (0.086 ± 0.007 and 0.258 ± 0.009 µg/g for control group and 0.149 ± 0.010 and 0.339 ± 0.013 µg/g for patient group, respectively for Ni and Cr, p < .001). ANCOVA test by removing the effects of age, gender and the baseline levels of Ni and Cr showed that changes in these ions in the scalp hair of both groups after one year were statistically significant. CONCLUSION: Due to the slightly elevated levels of Ni and Cr ions in the scalp hair of patients treated with fixed orthodontic appliances and considering the cytotoxic and allergic effects of these ions, changing the ingredients in fixed orthodontic appliances is suggested for the future.

Human serum miR-34a as an indicator of exposure to ionizing radiation.

Radiation exposure in industrial accidents or nuclear device attacks is a major public health concern. There is an urgent need for markers that rapidly identify people exposed to ionizing radiation (IR). Finding a blood-based marker is advantageous because of the ease of sample collection. This study was designed to test the hypothesis that serum miR-34a could serve as an indicator of exposure to IR. Therefore, 44 women with breast cancer, where radiotherapy was part of their therapeutic protocol, were investigated in this study. After demonstrating the appropriateness of our microRNA (miRNA) extraction efficiency and miRNA assay in human serum, we analyzed the miR-34a level in paired serum samples before and after radiotherapy. Fifty Gy X-ray irradiation in daily dose fractions of 2 Gy, 5 days per week, was used in this study. We demonstrated that IR significantly increased serum level of miR-34a. By measuring miR-34a in serum, we could distinguish irradiated patients with sensitivity of 65 % and specificity of 75 %. According to this study, serum miR-34a has the potential to be used as an indicator of radiation exposure.

Hyaluronic acid algorithm-based models for assessment of liver fibrosis: translation from basic science to clinical application.

BACKGROUND: The estimation of liver fibrosis is usually dependent on liver biopsy evaluation. Because of its disadvantages and side effects, researchers try to find non-invasive methods for the assessment of liver injuries. Hyaluronic acid has been proposed as an index for scoring the severity of fibrosis, alone or in algorithm models. The algorithm model in which hyaluronic acid was used as a major constituent was more reliable and accurate in diagnosis than hyaluronic acid alone. This review described various hyaluronic acid algorithm-based models for assessing liver fibrosis. DATA SOURCE: A PubMed database search was performed to identify the articles relevant to hyaluronic acid algorithm-based models for estimating liver fibrosis. RESULT: The use of hyaluronic acid in an algorithm model is an extra and valuable tool for assessing liver fibrosis. CONCLUSIONS: Although hyaluronic acid algorithm-based models have good diagnostic power in liver fibrosis assessment, they cannot render the need for liver biopsy obsolete and it is better to use them in parallel with liver biopsy. They can be used when frequent liver biopsy is not possible in situations such as highlighting the efficacy of treatment protocol for liver fibrosis.

Pre and post radiotherapy serum oxidant/antioxidant status in breast cancer patients: Impact of age, BMI and clinical stage of the disease.

AIM: In this study the effects of radiation therapy (RT) on serum oxidant/antioxidant status in breast cancer patients and the impact of age, BMI and clinical stage of the disease on the aforementioned variables were investigated. BACKGROUND: RT that is used for cancer treatment is dependent on the production of reactive oxygen species. MATERIALS AND METHODS: Eighty patients with breast cancer participated in this study and received RT at a dose of 50 Gy for 5 weeks. Blood samples were obtained in one day before and after the end of RT. Serum status of malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were analyzed by spectrophotometry or ELISA and selenium (Se) level were analyzed by atomic absorption spectrometry. Paired t-test was used for comparing pre and post radiotherapy data. RESULTS: Before and after the radiotherapy, a significant increase in MDA level was observed, while a significant decrease in GPx activity, SOD, TAS and Se levels were found (p < 0.05). The level of the CAT enzyme had no significant changes (p = 0.568). The results showed some changes in the status of TAS, SOD and GPx which are associated with age, BMI and clinical stage of the disease. CONCLUSION: It seems that RT would have the potential to cause variations in the status of antioxidant/oxidant system. Although, some changes in variables were observed by sub-classification of the age, BMI and the disease stage, but it seems that these changes are not necessarily dependent to them.

Starved human fibroblasts secrete acidic proteins inducing post re-feeding proliferation and in vitro cell migration: a potential tool for wound healing.

BACKGROUND INFORMATION: There are several reports indicating that starved fibroblasts show higher proliferation rates when re-fed with foetal bovine serum. We have evidence demonstrating that this phenomenon is related to secretory proteins which may be beneficial to wound healing. RESULTS: After re-feeding, 16 and 72 h serum-starved fibroblasts showed the highest and lowest proliferation rates, 1.59 and 0.51-fold difference compared to the non-starved control, respectively (P < 0.05). However, the latest value could be normalised by incubating cells with 16 h-starved fibroblast cell culture supernatant (16-SFS), prior to re-feeding. A strong correlation was found between total protein level in starved fibroblast culture supernatants and post re-feeding proliferation rates (r(2) = 0.90, P < 0.001). Two-dimensional gel electrophoresis analysis of 16-SFS confirmed the presence of proteins with relative molecular weights of 10-120 kDa and pI ranging from 4 to 6. A significant difference in calcium influx course was found between 16-SFS and the negative control (Dulbecco's Modified Eagle Medium) (P < 0.05). There was no significant difference in Ca(2+) concentrations after 1 h between non-starved controls and 16-SFS-treated fibroblasts. The scratch test demonstrated that the 16-SFS is able to induce fibroblast migration. CONCLUSIONS: We concluded that human starved fibroblasts secrete proteins that are able to induce post re-feeding cell proliferation and fibroblasts migration, probably through the induction of a sustained calcium influx. This is worth being considered as a potential tool for wound healing.

Effect of granulocyte colony-stimulating factor administration on tissue regeneration due to carbon tetrachloride-induced liver damage in experimental model.

The mechanism by which granulocyte colony-stimulating factor (G-CSF) could lead to the protection from liver injury is not well known. Therefore, the resolution of this role needs further basic and clinical experimental investigation. Acute liver injury was induced in rats by single intraperitoneal injection of a 0.50-mL/kg dose of carbon tetrachloride (CCl4 ). Granulocyte colony-stimulating factor or vehicle of 150 µg/kg was given immediately after intoxicating the liver by CCl4 . The animals were divided into four groups of twelve each. Administration of G-CSF caused a decrease in the activity of liver enzymes,  aminotransferases, compared with the untreated group.

SAR131675 exhibits anticancer activity on human ovarian cancer cells through inhibition of VEGFR-3/ERK1/2/AKT signaling pathway.

Vascular endothelial growth factor receptor-3 (VEGFR-3) is known to participate in tumorigenesis and lymphangiogenesis, and as such, has the potential to serve as a molecular target for cancer therapy. SAR131675 is a highly selective VEGFR-3 antagonist that has an inhibitive effect on lymphatic cell growth. However, the anticancer effects and underlying mechanisms of SAR131675 in ovarian cancer remain poorly understood. In this study, we investigated the pathological role of VEGFR-3, and the effects of SAR131675 on proliferation, cell cycle, migration, and apoptosis in ovarian cancer cells. Our results showed that the mRNA and protein of VEGFR-3 were expressed in OVCAR3 and SKOV3 ovarian cancer cells, and this receptor was activated following stimulation with 50 ng/ml VEGF-C Cys156Ser (VEGF-CS), a selective ligand for VEGFR-3. Enhancing VEGFR-3 phosphorylation by treatment of ovarian cancer cells with VEGF-CS resulted in increased levels of phosphorylated extracellular signal-regulated kinases 1/2 (ERK1/2) and AKT. Moreover, our data demonstrated that SAR131675 inhibited VEGF-CS-mediated proliferation, colony formation, and migration of cancer cells in a dose-dependent manner. In addition, inhibition of VEGFR-3 activation with SAR131675 significantly increased cell cycle arrest and promoted apoptosis in both OVCAR3 and SKOV3 cells. Mechanistically, SAR131675 effectively suppressed the VEGF-CS-induced phosphorylation of VEGFR-3 and its downstream effectors including activated ERK1/2 and AKT in ovarian cancer cells. Our results reveal an anticancer activity of SAR131675 on the growth and migration of ovarian cancer cells, which may be through inhibiting VEGFR-3/ERK1/2/AKT pathway. SAR131675 may serve as an effective targeted drug for ovarian cancer.

Expression Status of Rap1 Pathway-Related Genes in Liver Metastases Compared with Corresponding Primary Colorectal Cancer.

Understanding molecular networks of CRLM is an ongoing area of research. In this study, paired CRC tissue and adjacent noncancerous tissue from 15 non-metastatic CRC patients and paired CRC tissue and matched liver metastatic tissues from 15 CRLM patients along with their adjacent noncancerous tissues were evaluated. We assessed Rap1 pathway-related genes including NRAS, FGF-1, NGF, and KDR expression by qRT-PCR and their protein status by Western blot. In CRLM patients, NRAS, FGF1, and KDR mRNA and protein were expressed at higher levels in metastatic than in CRC primary tumor and adjacent noncancerous tissue (p < 0.05). In non-metastatic patients, NRAS, FGF1, KDR, and NGF gene expression did not differ between CRC primary tumor-and adjacent noncancerous tissue (p > 0.05). ROC curve analysis showed a reasonable diagnostic accuracy of NRAS, FGF1, KDR, and FGF for the discrimination of metastatic patients from non- metastatic ones on analysis of their primary tumors. The data suggest that further functional studies on Rap1-related genes' role in CRLM are needed. In conclusion, the present data broaden our knowledge about specific molecular characteristics of CRLM. An increased understanding of the molecular features of metastasis has the potential to create more successful treatment, or prevention, of metastasis, especially in multimodal primary tumor treatment.