NF-κB2 and RELB offer prognostic information in colorectal cancer and NFKB2 rs7897947 represents a genetic risk factor for disease development.

The Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) family of transcription factors plays an important role in immune responses and cancer development and progression. We have focused on NF-κB2 and RELB of the alternative pathway of NF-κB, which remains largely underexplored in colorectal cancer (CRC). We found that NF-κB2 and RELB protein levels were upregulated in tumour and surrounding stromal tissue compared to distant non-neoplastic tissue (NN) and associated stroma (p<0.001 in all associations). Moreover, low RELB protein expression was associated with decreased overall survival (p = 0.032). Lower RELB gene expression levels were observed in tumour compared to NN tissue (p = 0.003) and were associated with shorter time to progression (TTP) (p = 0.025). NF-κB2 gene expression levels were similar in tumour and NN tissue, but higher tumour levels were prognostic for improved survival (p = 0.038) and TTP (p<0.001). We also assessed the significance of two NF-κB2 genetic polymorphisms, rs12769316 and rs7897947. Both polymorphisms were associated with lymph node infiltration (p = 0.045 and p = 0.009, respectively). In addition, rs12769316 AA homozygotes relapsed less often compared to G allele carriers (p = 0.029). Moreover, rs7897947 allele frequencies differed significantly between CRC patients and healthy controls (p<0.001) and the minor allele (G) was associated with reduced risk for developing CRC (p<0.001, OR: 0.527, 95% CI: 0.387-0.717). In conclusion, the alternative NF-κB pathway appears deregulated in CRC. Moreover, NF-κB2 and RELB expression levels seem to be significant for the clinical outcome of CRC patients and rs7897947 appears to be a risk factor for CRC development.

Variant of BCL3 gene is strongly associated with five-year survival of non-small-cell lung cancer patients.

OBJECTIVES: BCL3, a known atypical IκB family member, has been documented to be upregulated in hematological malignancies and in some solid tumors, functioning as a crucial player in tumor development. Recently, rs8100239, a tag-Single Nucleotide Polymorphism (SNP) in BCL3 (T>A) has been identified, but there are no data regarding its involvement in non-small-cell lung cancer (NSCLC) initiation and progression. MATERIALS AND METHODS: To study the possible association of BCL3 with NSCLC, 268 patients and 279 healthy controls were genotyped for rs8100239. Moreover, BCL3 protein expression was also investigated in 112 NSCLC cases through an immunohistochemical analysis. RESULTS: NSCLC patients with AA genotype displayed significantly worse prognosis compared to T allele carriers (P<0.001), who had less frequent intermediate nuclear BCL3 expression (P=0.042). In addition, overexpression of BCL3 was detected in tumor specimens, compared to normal tissue (P<0.001). Furthermore, BCL3 protein levels were associated with five-year survival (P=0.039), maximum diameter of lesion (P=0.012), grade (P=0.002) and relapse frequency (P=0.041). CONCLUSIONS: The present study is the first to show a relationship between the genetic variation rs8100239 of BCL3 and cancer patients' survival. It also represents the first quantitative evaluation of BCL3 expression in NSCLC. Our findings indicate that rs8100239 may be considered as a novel prognostic indicator, demonstrating also the overexpression of BCL3 protein in NSCLC and implicating this pivotal molecule in the pathogenesis of NSCLC.

Association of functional VKORC1 promoter polymorphism with occurrence and clinical aspects of ischemic stroke in a Greek population.

Genetic factors are considered to play an important role in determining the susceptibility to the occurrence, clinical course, and functional outcome of an acute ischemic stroke (IS). Undercarboxylation of specific vitamin K-dependent proteins, due to genetic polymorphisms of VKORC1, can affect both vascular calcification and thrombogenicity. We sought to determine the association of VKORC1 -1639G > A polymorphism with IS incidence, age of onset, severity of disease, and functional outcome after an acute IS. VKORC1 -1639G > A polymorphism was determined in 145 consecutive patients with first ever IS and 145 age- and sex-matched control subjects of Greek Caucasian origin using PCR-RFLP. Stroke severity and functional outcome were assessed on admission and at one month after stroke, respectively. Frequency of VKORC1 -1639G > A genotypes did not differ between IS patients and controls (OR = 1.12, P = 0.51). Moreover, carriage of the A allele was not associated with age of stroke onset, severity of disease (Scandinavian stroke scale score 32.2 versus 32.9, resp., P = 0.96), or poor outcome at 1 month post-stroke (52.9 versus 64.4%, resp., P = 0.31). In conclusion, VKORC1 -1639G > A polymorphism is not a genetic determinant of IS occurrence, age of onset, severity, or functional outcome of disease in a Greek population.

Association of monocyte chemoattractant protein-1 -2518A>G polymorphism with occurrence, severity, and outcome in ischemic stroke.

Monocyte chemoattractant protein-1 (MCP-1) is implicated in promoting atherosclerotic diseases, including stroke. Therefore, several studies have investigated the association between variants of the MCP-1 gene and risk of atherosclerotic diseases. We sought to determine the occurrence of MCP-1 -2518A>G polymorphism in patients with ischemic stroke (IS), and studied its association with the severity of disease and functional outcome after an acute IS. One hundred and forty-five consecutive patients with first ever IS and 145 age- and sex-matched control subjects were recruited. Stroke severity and functional outcome were assessed on admission and at one month post-stroke, respectively. Genotyping for the MCP-1 -2518A>G polymorphism was performed by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). No significant difference in the frequency of MCP-1 -2518A>G genotypes between IS patients and controls was found, with OR = 0.69 (95 % CI 0.46-1.04, P = 0.08). Moreover, carriage of the G allele was not associated with stroke severity (Scandinavian stroke scale score 33.1 vs. 32.5, respectively, P = 0.71), or poor outcome at 1 month post-stroke (63.9 vs. 59.7 %, respectively, P = 0.61). In conclusion, we were unable to demonstrate a significant association of the MCP-1 -2518A>G gene polymorphism with IS occurrence, severity or functional outcome in a Caucasian population. However, larger studies are necessary to fully elucidate the role of this polymorphism in IS.

Association of endothelial nitric oxide synthase polymorphism G894T with functional outcome in acute stroke patients.

OBJECTIVES: Endothelial nitric oxide synthase (eNOS) G894T polymorphism has been previously associated with vascular diseases including stroke, but often with conflicting results. Experimental evidence suggests that some eNOS modalities may be important in preventing stroke, while others may be important during the dynamic inflammatory processes triggered at the acute phase of stroke. Thus, we examined the possible association of eNOS G894T variation with stroke severity and functional outcome. METHODS: One hundred and eight consecutive patients with first ever acute atherothrombotic or lacunar stroke, and 193 stroke-free subjects were recruited. Demographic data, medical history, and vascular risk factors were collected. Assessments for stroke severity and functional outcome were carried out on admission and at one month post-stroke, respectively. eNOS G894T genotypes were produced using a standard restriction-fragment-length polymorphism technique. RESULTS: eNOS G894T polymorphism genotypic distributions did not differ between stroke patients and controls, as well as between each stroke subtype and controls. Carriage of T allele (GT and TT genotypes versus GG) was not significantly associated with either stroke severity or functional outcome in the univariate analysis. However, after accounting for age, gender and stroke severity, the presence of T allele strongly predicted poor outcome [odd ratio 8·49 (95% confidence intervals 1·57-46·0)]. Further adjustments for other important confounders could not alter the above significant association. DISCUSSION: Carriers of the eNOS 894T allele are at increased risk for adverse outcome at one month post-stroke, independently of severity and other confounding factors. Therapeutic interventions targeting patients with this unfavorable mutation may be an appealing approach.

Functional inflammatory genotypes in ischemic stroke: could we use them to predict age of onset and long-term outcome?

Functional single-nucleotide polymorphisms (SNPs) of inflammatory cytokines have been previously related to the occurrence of an ischemic stroke (IS). We investigated whether five functional SNPs (i.e., TNF-α-308G>A, IL6-174G>C, IL12B 1188A>C, IL4-589C>T, and IL10-1082G>A) might be associated with the age of onset and 6-month outcome of an acute IS. A probe-free real-time PCR methodology was used to genotype 145 consecutively admitted cases with a first-ever IS. Simple Kaplan-Mayer and adjusted Cox regression analyses showed no association between inflammatory genotypes and the age of IS onset. IL6-174G>C, IL12B 1188A>C, IL4-589C>T, and IL10-1082G>A were not found to significantly contribute to the long-term outcome of the disease. However, carriage of the TNF-α-308 GG genotype was significantly associated with reduced odds for an adverse outcome. Larger studies are needed to confirm our results.

The survivin -31 snp in human colorectal cancer correlates with survivin splice variant expression and improved overall survival.

BACKGROUND: Survivin is involved in the regulation of cell division and survival, two key processes in cancer. The majority of studies on survivin in colorectal cancer (CRC) have focused on protein expression and less is known about the expression of survivin splicing variants or survivin gene polymorphisms in CRC. In the present study, the mRNA levels of the five known isoforms of survivin as well as survivin protein were assessed in matched normal and neoplastic colorectal tissue. Moreover, the 9386 C/T and -31 G/C polymorphisms were investigated. METHODS: Quantitative RT-PCR was used to assess mRNA levels in fresh/frozen tissue samples. Protein levels were immunohistochemically evaluated on formalin-fixed paraffin-embedded tissue sections. Individuals were genotyped using real time PCR. RESULTS: Expression of all 5 survivin splice variants as well as survivin protein was elevated in colorectal carcinomas compared to normal tissue. Specific splice variant expression differentially correlated with clinicopathological parameters. Furthermore, both snps correlated with splice variant levels or their ratios in colorectal carcinomas while the -31 G/C snp may be related to CRC development and improved overall survival. CONCLUSION: Our results support a role of survivin in colorectal carcinogenesis while the -31 G/C snp may constitute a marker of survival.

VEGF polymorphisms may be associated with susceptibility to colorectal cancer: a case-control study.

The vascular endothelial growth factor (VEGF) has a pivotal role in angiogenesis. VEGF levels appear to be influenced by single nucleotide polymorphisms (SNPs) of the VEGF gene. The aim of this study was to assess the importance of four VEGF SNPs in modulating susceptibility to colorectal cancer. We have genotyped 223 patients with colorectal cancer and 264 healthy individuals for the -2578C>A, -1498C>T, -634G>C and +936C>T VEGF SNPs using Taqman probes in polymerase chain reactions. The -2578 A, -1498 C and -634 G alleles were more frequently detected in CRC patients compared to healthy controls. Moreover, the haplotype -2578C/-1498T was less frequent in CRC patients while the -2578A/-1498C haplotype was significantly more frequent in patients compared to healthy controls. VEGF -2578C>A and -1498C>T SNPs and -2578/-1498 haplotypes appear to be associated with susceptibility to CRC.

The survivin -31 snp in human colorectal cancer correlates with survivin splice variant expression and improved overall survival.

BACKGROUND: survivin is involved in the regulation of cell division and survival, two key processes in cancer. The majority of studies on survivin in colorectal cancer (CRC) have focused on protein expression and less is known about the expression of survivin splicing variants or survivin gene polymorphisms in CRC. In the present study, the mRNA levels of the five known isoforms of survivin as well as survivin protein were assessed in matched normal and neoplastic colorectal tissue. Moreover, the 9386C/T and -31G/C polymorphisms were investigated. METHODS: quantitative RT-PCR was used to assess mRNA levels in fresh/frozen tissue samples. Protein levels were immunohistochemically evaluated on formalin-fixed paraffin-embedded tissue sections. Individuals were genotyped using real time PCR. RESULTS: expression of all 5 survivin splice variants as well as survivin protein was elevated in colorectal carcinomas compared to normal tissue. Specific splice variant expression differentially correlated with clinicopathological parameters. Furthermore, both snps correlated with splice variant levels or their ratios in colorectal carcinomas while the -31G/C snp may be related to CRC development and improved overall survival. CONCLUSION: our results support a role of survivin in colorectal carcinogenesis while the -31G/C snp may constitute a marker of survival.

Prion protein expression and the M129V polymorphism of the PRNP gene in patients with colorectal cancer.

The prion protein, PrP(C), is known mostly for its involvement in neurodegenerative spongiform encephalopathies. However, a role for this molecule in cancer is becoming increasingly recognized partly because it promotes cell proliferation and inhibits apoptosis. Moreover, the codon 129 polymorphism (M129V) of the PRNP gene (the PrP(C)-encoding gene) has been associated with neurodegenerative disease development and severity, while no information is available regarding its role in colorectal cancer (CRC) incidence and disease progression. We have previously reported that expression levels of PRNP may have a prognostic value in CRC, suggesting a role for the prion protein in CRC. The aim of this study was to investigate retrospectively the possible role of M129V and PrP(C) expression in patients with CRC. The M129V single nucleotide polymorphism was genotyped by real time polymerase chain reactions in 110 patients with CRC and 124 healthy donors. Moreover, protein expression was assessed by immunohistochemistry in 68 patients with CRC. Allele frequencies were similar in patients and healthy controls indicating that the M129V polymorphism is not a risk factor for CRC. Furthermore, it did not correlate with any clinicopathological parameters. By contrast, PrP(C) expression was highly elevated in neoplastic compared to normal tissue and differed depending on the primary site. Interestingly, protein levels were correlated with disease recurrence (P = 0.007). Conclusively, PrP(C) overexpression may constitute a prognostic marker for disease recurrence and potentially a new target for anticancer therapy. However, further studies are needed to evaluate prospectively the role of PrP(C) expression in patients with CRC.

Acute post-stroke adiponectin in relation to stroke severity, progression and 6 month functional outcome.

BACKGROUND: Circulating adiponectin (ADPN) has been inversely associated with the risk of coronary artery disease and ischemic stroke (IS), due to its anti-inflammatory and anti-atherosclerotic properties. Recent experimental studies have suggested that ADPN may as well exert cerebroprotective properties in brain ischemia, therefore modifying disease outcome. We investigated whether acute post-stroke ADPN in humans might be associated with disease severity, progression and outcome. METHODS: Serum ADPN was measured in 82 consecutive acute IS patients. Severity at presentation and stroke progression within the first week were evaluated according to internationally agreed definitions. Disability and functional outcomes were assessed on months 1, 3 and 6 using the modified Rankin scale (mRS) and Barthel index (BI). Additional data included information on infarct size, mortality, recurrent IS and mental state. RESULTS: Higher ADPN was indicative of greater disability (mRS) on month 1 (OR=1·141, 95% CI=1·012-1·286, p=0·031), but this result was not replicated using the BI. ADPN was not found to be associated with either stroke severity, clinical progression, infarct size, recurrent IS, mortality, mental state, disability or functional outcome during the 6 month follow-up. CONCLUSIONS: Despite previous experimental evidence, serum ADPN measured shortly after an acute IS in humans does not seem to reliably predict disease severity, progression or outcome. The concept that circulating ADPN may beneficially modify long-term outcome of an acute IS may not be the case for human stroke.