Relationship of serum levels of VEGF and TGF-β1 with radiosensitivity of elderly patients with unresectable non-small cell lung cancer

This study aimed to evaluate the relationship of serum levels of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) with radiosensitivity of elderly patients with unresectable non-small cell lung cancer (NSCLC) receiving three-dimensional conformal radiation therapy (3D-CRT). Fifty-eight elderly patients with unresectable NSCLC and 40 healthy controls were enrolled in this study. Serum levels of VEGF and TGF-β1 were detected by the enzyme-linked immunosorbent assay (ELISA) method before and after 3D-CRT. Clinical performances of serum VEGF and TGF-β1 levels in predicting radiosensitivity of NSCLC patients with 3D-CRT were evaluated. Serum VEGF and TGF-β1 levels of NSCLC patients were higher than those of health controls (all p?<?0.05). After 3D-CRT treatment, 41 patients achieved effective clinical response (complete response (CR)?+?partial response (PR)) and 17 patients were ineffective clinical response (stable disease (SD)?+?progressive disease (PD)). There was no significant difference in the VEGF and TGF-β1 levels between the effective and ineffective groups before 3D-CRT (all p?>?0.05). Serum levels of VEGF and TGF-β1 after 3D-CRT in the effective group were lower compared with the levels before 3D-CRT treatment (p?<?0.001 and 0.027, respectively). However, no significant differences in serum VEGF and TGF-β1 levels between before and after 3D-CRT in the ineffective group were observed (p?=?0.196 and 0.517, respectively). We observed significant differences in serum VEGF and TGF-β1 levels between the effective and ineffective groups after 3D-CRT (p?<?0.001 and 0.013, respectively). Sensitivity and specificity of VEGF combined with TGF-β1 in predicting radiosensitivity of NSCLC patients with 3D-CRT were 87.8 and 94.1 %, respectively. In conclusion, our results indicate that serum VEGF and TGF-β1 levels may accurately predict radiosensitivity of elderly patients with unresectable NSCLC receiving 3D-CRT.     

Altered expression of TGF-β1 and TGF-βR2 in tissue samples compared to blood is associated with food habits and survival in esophageal squamous cell carcinoma

In the transforming growth factor β (TGF-β) signaling pathway, TGF-β1 and TGF-β receptor 2 (TGF-βR2) are essential regulatory components which play an important role in different type of cancer. Expressions of TGF-β1 and TGF-βR2 were done by real-time qPCR in both biopsy and blood samples collected from esophageal squamous cell carcinoma (ESCC) patients (n = 76). The expression profiles were correlated with different lifestyle factors and clinicopathological parameters. Kaplan-Meier survival analysis and Cox regression analysis were performed to estimate survival and hazard outcomes of different parameters. TGF-β1 showed upregulation in 91% tissue samples (2.84 ± 1.34*) and 55% blood samples (2.43 ± 1.24*) whereas expression of TGF-βR2 showed downregulation in 89% tissue samples (0.27 ± 0.23*) and 75% blood samples (0.30 ± 0.26*). Among all the parameters, TGF-β1 expression is significant with histopathology grade, consumption of betel nut and smoked food whereas TGF-βR2 expression is significant only with dysphagia grade in both blood and tissue samples and while analyzing both male and female patients separately. Consuming alcohol and hot food, difference in tumor stage and metastasis were found to have statistically significant (P < 0.05) impact on survival and mortality of male patients while consuming hot food, tobacco, metastasis and TGF-βR2 expression in tissue level were found to associate with survival and mortality of female patients. Expression of both TGF-β1 and TGF-βR2 in tissue samples may be prospective biomarkers for screening of ESCC among the Northeast population. Survival outcomes and hazard analysis supports the importance of some clinicopathological and lifestyle factors on ESCC development, whereas expression study depicts association of change in expression of the studied genes in ESCC patients.*Mean fold change.     

Association of -308 TNF-α promoter polymorphism with clinical aggressiveness in patients with head and neck squamous cell carcinoma

Genetic polymorphisms in the promoter region of the tumour necrosis factor-α (TNF-α) gene are involved in the regulation of the expression levels of its cytokine. Besides, these polymorphisms have been associated with the clinical behaviour of cancer. We investigated the −308 promoter region polymorphisms of the TNF-α gene and its association with the clinicopathological factors of a head and neck squamous cell carcinoma (HNSCC) sample. Furthermore, we analysed the impact of all the variables on the overall survival of patients. A sample of HNSCC (n = 89) was evaluated. Clinicopathological factors and overall survival data were gathered. The TNF-α gene was analysed by using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). Data analyses were performed by using bivariate and multivariate statistical tests. Significance was set at p < 0.05. HNSCC subjects carrying the A allele (GA/AA) exhibited associations with poor performance status (OR = 2.82, p = 0.039), lesions located on posterior areas (OR = 4.02, p = 0.002), and large-size tumours (OR = 2.91, p = 0.015). Subjects carrying only AA genotype exhibited association with poor performance status (OR = 6.667, p = 0.007). A worse overall survival was noted in subjects with large tumours (OR = 4.87, p = 0.005) and locoregional metastatic disease (OR = 2.50, p = 0.018). Our data suggests that the presence of the A allele/AA haplotype in HNSCC individuals might contribute to the higher clinical aggressiveness of malignant disease.     

Immunohistochemical study of β-catenin and functionally related molecular markers in tongue squamous cell carcinoma and its correlation with cellular proliferation

β-catenin plays an important role in the maintenance of cell adhesion and is a key component of the Wnt signaling pathway. However, little is known about its prognostic significance or its role in tumor progression in tongue squamous cell carcinoma (SCC). This study conducted an immunohistochemical analysis of the expression of β-catenin. Moreover, its possible correlation with clinical parameters and with the expression of the functionally related molecular markers cyclin D1 and p53 was evaluated in 50 cases of tongue SCC and 10 cases of normal tongue epithelium. The ki-67 labeling index (LI) was also examined to evaluate cellular proliferation. Our results showed a higher frequency of abnormal β-catenin expression, positive cyclin D1 and p53 expression, and a significantly higher ki-67 LI in the tongue SCC samples compared with normal tongue epithelium (P<0.05). Abnormal β-catenin and a higher ki-67 expression was significantly associated with moderately or poorly differentiated carcinoma (P<0.05). Cyclin D1-positive immunostaining showed a statistically significant association with lymph node metastasis (P<0.05). Furthermore, the abnormal expression of β-catenin significantly correlated with a higher ki-67 LI and p53 expression (P<0.05); however, there was no correlation with cyclin D1 expression (P>0.05). Taken together, our results suggest that abnormal β-catenin expression is related to the impaired cellular differentiation and proliferation involved in tumor progression in tongue SCC.     

Hypomethylation of LINE-1, and not centromeric SAT-α, is associated with centromeric instability in head and neck squamous cell carcinoma

Background

Head and neck squamous cell carcinoma (HNSCC) is a tumour type that generally carries very complex chromosomal aberrations. An interesting feature is the elevated occurrence (58?%) of whole arm translocations and isochromosomes, resulting from breakage and illegitimate recombination in centromeric or pericentromeric regions. We hypothesized that alterations in DNA methylation may play a role in the breakage of centromeric repeat sequences in these tumours.

Methods

We studied the DNA methylation status of global repeats (LINE-1), subtelomeric repeats (D4Z4) and centromeric repeats (SAT-??) in relation to centromeric instability in a series of HNSCC cancer cell lines and primary tumours. We analysed the methylation status by pyrosequencing and the chromosomal aberrations by microarray CGH.

Results

We found a significant association between centromeric instability and hypomethylation of LINE-1, but not D4Z4 and SAT-??.

Conclusion

These data suggest that centromeric instability is associated with genomic DNA hypomethylation only when occurring at specific DNA repeat sequences.     

Synergistic effects of imatinib and carboplatin on VEGF, PDGF and PDGF-Rα/ß expression in squamous cell carcinoma of the head and neck in vitro

Head and neck squamous cell carcinoma (HNSCC) is an aggressive epithelial malignancy. The development of new treatment modalities in order to improve long-term survival of patients with HNSCC is imperative. Numerous studies have demonstrated that carcinogenesis and tumor cell dissemination is influenced by the tumor microenvironment. The protein-kinase-receptors (PTKs) are essential elements of the intracellular signal transduction pathway and regulate cell growth, development and apoptosis. Cell proliferation, migration, induction of tumor vascularization and carcinogenesis, invasion is regulated by a variety of angiogenic factors, such as PDGF (platelet-derived growth factor), VEGF (vascular endothelial growth factor) and their respective tyrosine kinase receptors (PDGF-R and VEGF-R). They present promising targets for anti-cancer therapy through abrogation of impaired signaling pathways. Indeed, imatinib, a small molecule drug targeting these protein kinases, has antiproliferative effects in several cancer types. The purpose of this study was to investigate the potential synergism of imatinib and carboplatin on the expression of PDGF, PDGF-R α/? and VEGF in different HNSCC cell lines. Several tumor cell lines were subjected to increasing concentrations of carboplatin (3 and 7.5 μmol/l) and imatinib (18 and 30 μmol/l) and ELISA, immunohistochemical methods and RQ-PRC after 48, 72, 120 and 240 h were used to assess their expression levels. While PDGF-Rα/? expression was unimpaired at lower imatinib concentrations (18 μmol/l), PDGF-Rα/? expression was suppressed at 30 μmol/l, and suppression was enhanced by the presence of carboplatin. By RQ-PCR, a significant reduction of PDGF-Rα/? expression was detected (p<0.5). We observed explicit significant reduction in VEGF levels with increasing concentrations of imatinib and with the combination of the two chemotherapeutic drugs (p<0.5). We report for the first time evidence of synergism of imatinib and carboplatin in suppressing VEGF, PDGF and PDGF-Rα/? expression in HNSCC.     

Expression and correlation of Lewis y antigen and TGF-β1 in ovarian epithelial carcinoma

Lewis y is a difucosylated oligosaccharide carried by glycoconjugates on the cell surface. Elevation of Lewis y is frequently observed in epithelial-derived cancers. This study aimed to detect the expression and clinical significance of the Lewis y antigen and TGF-β1 (transforming growth factor β1) in ovarian epithelial tumors, and to evaluate the correlation between them. Immunohistochemical staining was used to detect the expression of Lewis y antigen and TGF-β1 in 60 cases of ovarian epithelial malignant tumors, 20 cases of borderline ovary tumors, 20 cases of benign ovary tumors and 10 cases of normal ovarian tissues. An immunofluorescence double labeling method was also used to detect the correlation between Lewis y antigen and TGF-β1. The positive rates of Lewis y antigen in ovarian epithelial cancer tissues was 88.33%, significantly higher compared to those of borderline ovarian tumors (60.00%) (P<0.05), benign ovarian tumors (35.00%) (P<0.01) and normal ovarian tissues (0%) (P<0.01). Its expression was not associated with clinical parameters; the positive rates of TGF-β1 in ovarian epithelial cancers were 78.33%, significantly higher compared to those of benign ovarian tumors (65.00%) (P<0.05) and normal ovarian tissues (40.00%) (P<0.05); the positive rates of the TGF-β1 and Lewis y were not associated with metastasis of lymph nodes and histological types, differentiation degree and clinical stage (P>0.05). Expression of Lewis y antigen and TGF-β1 was significantly positively associated with epithelial carcinoma. Close correlation between Lewis y, TGF-β1 and ovarian cancer was observed. Altered expression of Lewis y antigen may cause changes in TGF-β1 expression. Lewis y can increase the growth of ovarian cancer cells and the invasion ability by promoting TGF-β1 abnormal expression and by promoting angiogenesis and a change in its signal transduction pathway. This study provides theoretical evidence for the development of ovarian cancer biological treatments.     

Immunohistochemical study of VEGF expression in oral squamous cell carcinomas: correlation with the mTOR-HIF-1α pathway

The aim of this study was to clarify the relationship between vascular endothelial growth factor (VEGF) expression and clinicopathological factors in oral squamous cell carcinoma (OSCC). We also examined the correlation between the VEGF expression and the mammalian target of rapamycin (mTOR)-hypoxia inducible factor-1α (HIF-1α) pathway. Formalin-fixed paraffin-embedded tissues from 120 OSCC cases and 10 samples of normal mucosa were stained immunohistochemically for VEGF-A, VEGF-C, p-mTOR and HIF-1α proteins. VEGF-A and VEGF-C protein expression was detected in 76 out of 120 (63%) and 81 of 120 (67.5%) OSCCs, respectively, and their expression was significantly higher in primary OSCC than in normal oral mucosa. VEGF-A expression was significantly associated with the tumor stage and age. VEGF-C expression was significantly associated with the cancer cell invasion. The cases with combined p-mTOR+/HIF-1α(+)/VEGF-A(+) expression had a significantly higher tumor stage and invasion grade, and combined p-mTOR+/HIF-1α(+)/VEGF-C(+) expression was significantly associated with tumor stage, regional lymph node metastasis and invasion grade. In a survival analysis, no obvious correlation was observed with any of the immunohistochemical results. This study indicated that the mTOR-HIF-1α-VEGF pathway affects the progression of OSCC, and inhibition of this pathway may be useful for the treatment of OSCC.     

Oestrogen receptors and peripheral serum levels of oestradiol-17β in patients with mammary carcinoma

Oestradiol receptors were determined by isoelectric focusing in cytosol from breast cancer tissue from 12 pre- and 70 postmenopausal patients. Simultaneous determination of the peripheral serum levels of oestradiol-17β was carried out in the 12 premenopausal and 57 of the postmenopausal patients. At oestradiol-17β levels below 150 pM the receptor values ranged from below 0.01 to 7.19 fmole bound [³H] oestradiol-17β/μg of DNA. At oestradiol-17β levels above 150 pM only low receptor values (<0.25 fmole bound [³H] oestradiol-17β/μg of DNA) were recorded. The results indicate an influence of the endogenous oestrogen on the receptor assay probably due to blockade of the hormone binding site. It is proposed that an endocrine evaluation of the patient should be performed together with the oestrogen receptor assay.     

DNA polymerase β mutations and survival of patients with esophageal squamous cell carcinoma in Linzhou City,China

Linzhou City in northern China has a high incidence of esophageal squamous cell carcinoma (ESCC). This study retrospectively analyzed the data of 231 cases with ESCC collected from 1998 to 2012. Mutations of DNA polymerase β (polβ) gene in the ESCC samples from patients in Linzhou City were examined by amplifying polβ cDNA by RT-PCR followed by cloning and sequencing. Mutations in polβ were found in 105 cases (45.9 %). Nine types of mutations were identified in the polβ cDNA; the most common were 177–234 nt deletion (11.3 %), 462 nt G?→?T (9.1 %), and 648 nt G?→?C (6.9 %). Mutations in polβ appeared to be associated with TNM status (P?=?0.048). Follow-up data was used for survival analysis. The overall 5-year survival rate of the 231 patients was 37.4 %; the rate for patients with wild-type (WT) polβ was 41.8 %. Compared with the WT polβ group, the median survival for patients with specific mutations (177–234 nt deletion, 462 nt G?→?T, or 613 nt A?→?T) was significantly shorter (all P?=?0.000), and the 5-year survival rate decreased to 0 %. Patients with the 648 nt G?→?C mutation had improved survival (P?=?0.000) with a 5-year survival rate of 100 %. Our results identified nine types of mutations within polβ cDNA in ESCC patients with four mutations related to patient survival.     

TNF-α and IL-10 promoter polymorphisms, HPV infection, and cervical cancer risk

Although the implication of genetic factors in cervical cancer development remains to be elucidated, accumulative epidemiological evidence suggests that polymorphisms of cytokine genes may be involved in the etiology of cervical carcinoma. Tumor necrosis factor alpha (TNF-??) and interleukin-10 (IL-10) are two multifunctional cytokines implicated in inflammation, immunity, and cellular organization, and were proposed to play important roles in cancer biology. In order to determine whether IL-10 -1082 (G/A) and TNF-?? -238 (G/A) and -308 (G/A) polymorphisms are associated with susceptibility to cervical cancer, a case?Ccontrol study of 122 cancer patients and 176 healthy controls was conducted. Cervical samples were genotyped for both TNF-?? polymorphisms by PCR-RFLP assay. SNP-1082 from IL-10 gene was genotyped using pyrosequencing technology. The association between cervical cancer risk and the studied SNPs was evaluated by logistic regression. Under univariate analysis, none of these polymorphisms appeared associated with susceptibility of cervical cancer development or HPV infection. However, individuals carrying heterozygous genotype for TNF-?? -238 polymorphism seem to be at lower risk for cervical cancer development, with borderline significance (OR?=?0.42, P?=?0.069), as well as those carrying heterozygous genotypes for IL-10 and TNF-?? -238 (OR?=?0.40, P?=?0.08). In conclusion, these results suggest a potential effect of TNF-?? -238 G/A in the reduction of cervical cancer risk in Argentine women, but not TNF-?? -308 or IL-10. Larger studies are needed to fully understand the genetic predisposition for the development of cervical cancer.     

Bone marrow mast cell density correlates with serum levels of VEGF and CXC chemokines ENA-78 and GRO-α in multiple myeloma

Angiogenesis is a crucial process in growth and progression of multiple myeloma (MM). Mast cells (MCs) play an important role in MM angiogenesis. Various angiogenic mediators secreted by MCs regulate endothelial cell proliferation and function. Among them, ELR⁺ CXC chemokines, such as growth-related oncogen-alpha (GRO-α) and epithelial neutrophil activating protein-78 (ENA-78), have been described as potential mediators in regulation of angiogenesis. The purpose of the study was to quantify MCs in bone marrow (BM) biopsies of MM patients, expressed as MC density (MCD), and correlate it with serum concentrations of vascular endothelial factor (VEGF), GRO-α, ENA-78. Fifty-four newly diagnosed MM patients and 22 healthy controls were studied. Tryptase was used for the immunohistochemical stain of MCs. VEGF, GRO-α, and ENA-78 were measured in sera by ELISA. MCD and serum levels of GRO-α, ENA-78, and VEGF were significantly higher in MM patients compared to controls (p?<?0.001 in all cases). MCD was significantly increasing with increased stage of the disease (p?<?0.001). Furthermore, significant correlations were found between MCD with VEGF, GRO-α, and ENA-78. These findings support that MCs participate in the pathophysiology of MM and is implicated in the angiogenic process and disease progression.     

CRP, TNFα, IL-1ra, IL-6, IL-8 and IL-10 in blood serum of colorectal cancer patients

Blood serum cytokines: TNFalpha, IL-1ra, IL-6, IL-8, IL-10 as well as CRP were investigated in patients with colorectal cancer, prior treatment and 1, 10 and 42 days after surgery. There was an increase of the levels of CRP, IL-6 and IL-10 in most patients 24 hours after surgery. The levels of IL-1ra were elevated in patients in stage C and in several patients in stage B of the disease and there was a decrease of circulating TNFalpha in stage B patients. On day 10 and 42 after surgery, the levels of cytokines followed various patterns.     

Prognostic value of β-catenin,c-myc,and cyclin D1 expressions in patients with esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma (ESCC) is one of the most frequently diagnosed malignant tumors in North China. We have identified that Wnt2/β-catenin pathway is activated in ESCC cells and sodium nitroprusside (SNP) and siRNA against β-catenin not only inhibit the expressions of β-catenin and its major downstream effectors including c-myc and cyclin D1 but induce cell cycle arrest and apoptosis. The purpose of the present study was to analyze the relationship between pathological parameters including invasion depth and lymph node metastasis and the expressions of β-catenin, c-myc, and cyclin D1 in order to evaluate their values of prognosis in patients with ESCC. The expressions of β-catenin, c-myc, and cyclin D1 were detected immunohistochemically in the resected cancer tissues from 40 patients with ESCC. The β-catenin expression was reduced in 22 (55.0%) patients, which was closely correlated with invasion depth (P = 0.023) and lymph node metastasis (P = 0.003). There was the positive c-myc expression in 21 (52.5%), which was significantly correlated with invasion depth (P = 0.009) and lymph node metastasis (P = 0.001). Furthermore, the results of survival rates analyzed by Kaplan–Meier curve revealed that patients with the reduced expression of β-catenin had a poorer prognosis than those with the preserved expression (P = 0.031), and patients with the positive expression of c-myc also had a significantly poorer prognosis than those with the negative expression (P = 0.008). These findings demonstrate that β-catenin pathway plays a crucial role in the progression of ESCC, suggesting that both β-catenin and c-myc may be used as markers for predicting the prognosis of patients with ESCC.