Cancer Biomarkers - Volume 17, issue 2

Authors: Enein, Azza A. Aboul | Rahman, Hala A. Abdel | Sharkawy, Nahla El | Elhamid, Samah Abd | Abbas, Sonia M.A. | Abdelfaatah, Rafaat | Khalil, Mohamed | Fathalla, Lamiaa A.

Article Type: Research Article

Abstract: BACKGROUND: CD99 was first isolated as an antigen on the T acute lymphoblastic leukemia cells. It has been shown to participate in T cell adhesion and is widely expressed on a variety of hematopoietic and non-hematopoietic cell types. AIM OF WORK: Detection of the expression pattern of CD99 on leukemic and normal T cells and assessing the possibility of its use as a tool for the diagnosis and monitoring of T-ALL cases. METHODOLOGY: We used flow cytometry technique to determine the expression level of CD99 in 62 newly diagnosed T-ALL patients. Patients were …followed up for the presence of minimal residual disease on day 15 and day 42 post-therapy. 20 age and sex matched healthy controls were enrolled in our study. RESULTS: CD99 was expressed in all T-ALL patients, with a higher median expression level when compared to controls (58.5% versus 1.38%, p< 0.001). On day 42 post-therapy, 100% of follow up patients who had initial CD99 expression ≤ 50% had no minimal residual disease, while only 45.5% of those who had initial CD99 expression > 50% had no minimal residual disease (P= 0.03). There was no significant influence of CD99 expression on the 1-year overall survival probability (P= 0.82). CONCLUSION: CD99 could be used to complement current strategy relying on TdT for diagnosis and monitoring of minimal residual disease during the post-therapy follow up of T-ALL patients. Further studies are needed to confirm these findings. Show more

Keywords: CD99, T acute lymphoblastic leukemia, disease free survival

DOI: 10.3233/CBM-160608

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 117-123, 2016

Authors: Akers, Johnny C. | Ramakrishnan, Valya | Yang, Isaac | Hua, Wei | Mao, Ying | Carter, Bob S. | Chen, Clark C.

Article Type: Research Article

Abstract: BACKGROUND: Tumor specific genetic material can be detected in extracellular vesicles (EVs) isolated from blood, cerebrospinal fluid (CSF), and other biofluids of glioblastoma patients. As such, EVs have emerged as a promising platform for biomarker discovery. However, the optimal procedure to transport clinical EV samples remains poorly characterized. METHODS: We examined the stability of EVs isolated from CSF of glioblastoma patients that were stored under different conditions. EV recovery was determined by Nanoparticle tracking analysis, and qRT-PCR was performed to determine the levels of miRNAs. RESULTS: CSF EVs that were lyophilized and stored …at room temperature (RT) for seven days exhibited a 37-43% reduction in EV number. This reduction was further associated with decreased abundance of representative miRNAs. In contrast, the EV number and morphology remained largely unchanged if CSF were stored at RT. Total RNA and representative miRNA levels were well-preserved under this condition for up to seven days. A single cycle of freezing and thawing did not significantly alter EV number, morphology, RNA content, or miRNA levels. However, incremental decreases in these parameters were observed after two cycles of freezing and thawing. CONCLUSIONS: These results suggest that EVs in CSF are stable at RT for at least seven days. Repeated cycles of freezing/thawing should be avoided to minimize experimental artifacts. Show more

Keywords: CSF, EV, exosome, stability, lyophilization, freeze thaw

DOI: 10.3233/CBM-160609

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 125-132, 2016

Authors: Bagci, Binnur | Sari, Musa | Karadayi, Kursat | Turan, Mustafa | Ozdemir, Ozturk | Bagci, Gokhan

Article Type: Research Article

Abstract: BACKGROUND: Colorectal cancer is a serious disease that causes significant morbidity and mortality in developed countries. Genetic changes, such as mutations in proto-oncogenes and DNA repair genes, and loss of function in the tumor suppressor genes cause colorectal cancer development. Abnormal DNA methylation is also known to play a crucial role in colorectal carcinogenesis. OBJECTIVE: In this study, frequencies of KRAS and BRAF mutations, promoter hypermethylation profiles of SFRP2, DAPK1, MGMT, HIC1 and p16 genes, and possible associations between hypermethylation of these genes and KRAS and BRAF mutations were aimed to find out. METHODS: …Ninety three colorectal cancer tissues and 14 normal colon mucosas were included in the study. Common twelve KRAS gene mutation were investigated with using reverse-hybridization strip assay method. BRAF V600E mutations were investigated with RFLP method. Hypermethylation status of five tumor suppressor genes were detected by using reverse-hybridization strip assay method after bisulfite modification of DNA. RESULTS: KRAS and BRAF mutation frequencies were determined as 54.84% and 12.9%, respectively. Promoter hypermethylation frequencies of tumor suppressor genes SFRP2, DAPK1, MGMT, HIC1 and p16 were determined as 66.7%, 45.2%, 40.9%, 40.9% and 15.1%, respectively. Statistically significant associations were found between BRAF mutation and SFRP2 and p16 tumor suppressor genes hypermethylation (SFRP2; p= 0.005, p16; p= 0.016). Compared to rectum, SFRP2 (p= 0.017) and MGMT (p= 0.013) genes have statistically significantly higher promoter hypermethylation in colon. CONCLUSIONS: Results of the current study have confirmed that KRAS mutations and SFRP2 hypermethylation can be used as genetic markers in colorectal cancer. Show more

Keywords: Colorectal cancer, hypermethylation, oncogene, tumor suppressor gene, KRAS, BRAF, SFRP2, DAPK1, MGMT, HIC1, p16

DOI: 10.3233/CBM-160624

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 133-143, 2016

Authors: Kaigorodova, Evgeniya V. | Zavyalova, Marina V. | Bychkov, Vyacheslav A. | Perelmuter, Vladimir M. | Choynzonov, Evgenii L.

Article Type: Research Article

Abstract: BACKGROUND: The small heat shock protein 27 kDA (Hsp27) acts as an ATP-independent chaperone in protein folding, but is also implicated in architecture of the cytoskeleton, cell migration, metabolism, cell survival, growth/differentiation, mRNA stabilization, and tumor progression. OBJECTIVE: To study the intracellular localization of phosphorylated and non-phosphorylated forms of Hsp27 in squamous cell carcinoma of the larynx (SCCL) and to evaluate their relationship with regional lymphatic metastasis and overall five-year survival. METHODS: Tumor biopsies of larynx tissue were collected from 50 patients who were between the ages of 30 to 80 years and …had a confirmed diagnosis of squamous cell carcinoma of the larynx. Immunohistochemistry was used to determine the intracellular localization of the phosphorylated and non-phosphorylated forms of Hsp27. RESULTS: The study revealed that the Hsp27 chaperone was expressed in both the cytoplasm and the nucleus of tumor cells in SCCL. The biopsies of patients with lymph node metastases showed significantly higher expression of the phosphorylated and unphosphorylated forms of Hsp27 in the nucleus compared to those of patients without lymph node metastases. At the same time, the cytoplasmic expression of Hsp27 in these patients did not differ statistically. Analysis of the overall five-year survival rates showed that negative Hsp27 expression in the nucleus of tumor cells is associated with the survival rate of patients with SCCL. CONCLUSION: The nuclear expression of phosphorylated and unphosphorylated forms of Hsp27 is a molecular marker of unfavorable squamous cell carcinoma of the larynx associated with lymphogenous metastasis and decreased total five-year survival. Show more

Keywords: Larynx cancer, heat shock protein 27, phospho S78-Hsp27, prognostic markers, immunohistochemistry

DOI: 10.3233/CBM-160625

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 145-153, 2016

Authors: Osama, Amany | Sabry, Dina | Hassany, Sahar M. | Abdelmoneim, Soha Saoud | Sabry, Abeer

Article Type: Research Article

Abstract: AIMS: The study aimed to investigate the quantitative expression of NANOG, p38 α , NCF2, ELF and TGF-β genes in patients with colorectal adenocarcinoma, adenoma and normal colonic tissue and their correlation with SIRT-1 protein level expression. METHOD: This study enrolled one hundred sixty seven patients; group A: 87 patients with colonoscopic findings of no adenoma or adenocarcinoma and group B: 80 patients with colorectal mass. Consecutive colonoscopic examinations were conducted, and tissue samples were taken from the colonic lesions/masses. Total RNA was isolated and mRNA expression level of NANOG, mitogen activated p38α , Neutrophil …Cytosol Factor 2 (NCF2), Embryonic Liver Fodrin (ELF) and Transforming Growth Factor Beta (TGF-β) genes were quantified by qRT-PCR. Sirt-1 protein expression level was assessed by quantitative western blot. RESULTS: There were significantly high level of mRNA transcripts expression of the genes studied in patients with adenocarcinoma and adenoma compared with normal tissue (P value < 0.01), NANOG, NCF2, ELF and TGF-β at a cut of > 0.314, > 0.392, 0.349 and 0.333 respectively showed sensitivity (96.5%, 98.8%, 95.3%, 98.8%) and specificity of (95.3%, 92.6%, 89.5%, 93.8%) respectively in diagnosing colonic adenocarcinoma. Sirt-1 protein level was significantly highly expressed in colorectal adenocarcinoma compared to normal and adenoma colonic tissue and positively correlated with NANOG. CONCLUSION: Over expression of NANOG, p38α , NCF2, ELF and TGF-β genes in both cases of adenocarcinoma and adenoma could have a diagnostic value. SIRT-1 and NANOG are high correlated biological markers for diagnosis and prognosis follow up in patients with adenocarcinoma. Show more

Keywords: NANOG, p38α , NCF2, ELF, TGF-β , SIRT-I, colon adenocarcinoma

DOI: 10.3233/CBM-160626

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 155-163, 2016

Authors: Xia, Haifeng | Shen, Ji | Chen, Shaomu | Huang, Haitao | Xu, Yaozeng | Ma, Haitao

Article Type: Research Article

Abstract: BACKGROUND: The prognostic value of vascular endothelial growth factor C (VEGF-C) in patients with esophageal cancer (EC) remains controversial. The aim of this meta-analysis was to clarify the association of VEGF-C with survival in EC patients. METHODS: We performed a meta-analysis that included eligible studies to expound the effect of VEGF-C in EC survival. Eligible studies published until November 2015 was identified using available databases. STATA 12.0 was performed in this meta-analysis. RESULTS: We identified 13 studies, including 1203 patients, in this meta-analysis. The combined hazard ratio of 1.70 (95% CI, 1.43-2.03, P …< 0.001) shows that VEGF-C overexpression was significantly correlated with poor overall survival in EC patients. Furthermore, the results suggested a significant relationship between VEGF-C expression and overall survival was also showed in studies with patient source, patient number ≥ 70, methods detecting VEGF-C by reverse transcription PCR (RT-PCR) or ELISA and histology type. Moreover, combined odds ratio of VEGF-C displayed that VEGF-C overexpression was significantly association with stage, depth of tumor invasion, lymph node status and metastasis of EC (P < 0.05). However, it has no correlation with differentiation degree of EC (P > 0.05). CONCLUSION: VEGF-C overexpression shows an unfavorable prognosis for EC patients. Show more

Keywords: VEGF-C, prognosis, esophageal cancer, meta-analysis

DOI: 10.3233/CBM-160627

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 165-170, 2016

Authors: Lu, Lin | Shen, Yuan | Tseng, Kuo-Fu | Liu, Wenlian | Duan, Hui | Meng, Wei

Article Type: Research Article

Abstract: BACKGROUND: Endometrial cancer (EC) is a common female malignancy. Most patients were diagnosed at an early stage with a favorable prognosis. But more than 30% patients were high risk at III/IV stage with invading deep into the myometrium and progressively lead to local or extra pelvic metastasis. Urothelial cancer-associated 1 (UCA1) had been reported as the oncogenic long non-coding RNA in many tumors, but the role of UCA1 in EC is still unclear. METHOD: QRT-PCR was used to analysis the level of UCA1 in the proliferative endometrium, primary EC tissue and lymph node metastasis tissue of …EC. According to the QRT-PCR results of primary EC tissue, survival curves were made using the Kaplan-Meier method, and the log rank test was used to analyze the differences of clinicopathological characteristics and survival between the low expression and high expression group of UCA1 in EC patients. Moreover we knocked down the expression of UCA1 in EC cell lines HTB-111 and Ishikawa, and detected the migration and invasion ability of them with wound healing assay and transwell assay. RESULTS: The expression level of UCA1 using QRT-PCR method in lymph node metastasis tissue was the highest than that in the proliferative endometrium and primary EC tissues (1.15 ± 0.23, 3.23 ± 1.06 vs. 6.42 ± 1.46, P < 0.0001). For the primary EC tissue, the median fold change of UCA1 was used as a cutoff value. High UCA1 expression was observed to be closely correlated with lymph node metastasis (P = 0.008), distant metastasis (P = 0.003), grade (P = 0.009), advanced TNM stage (P = 0.031) and vessel invasive (P = 0.032). The 5-year overall survival rate in the high expression group was 41.7%, compared with 72.7% in the low expression group (P = 0.023). After silencing the UCA1, the migration and invasion ability of EC cell lines reduced significantly. CONCLUSION: Our findings provide the convincing evidence that the UCA1 plays an important role in the metastasis of EC and may serve as a novel molecular marker to predict the aggressive tumor progression and unfavorable prognosis of EC patients. Show more

Keywords: Endometrial cancer, Long noncoding RNA, UCA1, metastasis

DOI: 10.3233/CBM-160628

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 171-177, 2016

Authors: Yang, Liu | Wang, Tiejun | Zhang, Jun | Liu, Zhonghao | Wang, Xuxia

Article Type: Research Article

Abstract: BACKGROUND: BTB/POZ domain-containing protein 7 (BTBD7) is recognized as a regulatory gene that regulates epithelial cell dynamics and branching morphogenesis. It is also reported for regulating epithelial-mesenchymal transition (EMT) molecules and involved in the process of invasion and metastasis of lung cancer and hepatocellular carcinoma. Slug is a transcriptional factor of EMT which plays a crucial role in the process of primary salivary adenoid cystic carcinoma (SACC). However, the role of BTBD7 in SACC and the correlation with Slug have not been identified. This study investigated the expression of BTBD7 and correlation with Slug, as well as the prognostic …significance of BTBD7 in SACC. METHODS: The expression of BTBD7 and Slug were examined in ACC-LM and ACC-83 cell lines and immunohistochemically in paraffin embedded tissue specimens from 66 primary SACC patients. Statistical analyses were performed to evaluate the correlation between BTBD7 expression and Slug expression and the prognostic significance of BTBD7 expression. RESULTS: BTBD7 protein expression was initially verified in ACC-LM and ACC-83 cell lines. The positive rate of BTBD7 expression was 62.1% in SACC to 20% in normal salivary tissues comparatively. BTBD7 expression was significantly correlated with Slug expression in SACC (P< 0.05). Increased BTBD7 expression was significantly associated with the TNM stage, tissue typing, distant metastasis and patients' poor clinical outcome. CONCLUSIONS: Positive expression of BTBD7 in SACC could play an important role in the development of cancer and may serve as a favorable predictor for diagnosis and poor prognosis of patients. Show more

Keywords: BTBD7, Slug, salivary adenoid cystic carcinoma, prognosis

DOI: 10.3233/CBM-160629

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 179-185, 2016

Authors: Zhang, Kaijiong | Luo, Zhenglian | Zhang, Yi | Zhang, Li | Wu, Lichun | Liu, Lian | Yang, Jie | Song, Xiaoyu | Liu, Jinbo

Article Type: Research Article

Abstract: BACKGROUND: Long non-coding RNA (lncRNA) H19 has been well studied playing an important role in breast cancer (BC) progress and the expression of H19 may service as a diagnostic target for BC. However, it is unclear if circulating lncRNA H19 could as a potential biomarker for BC diagnosis and monitor. OBJECTIVE: The objective of our study was to determine whether plasma lncRNA H19 could be used as biomarkers for the screening and early diagnosis of breast cancer. METHODS: In this study, we carried out a quantitative real-time PCR (qRT-PCR) method to examine the …expression levels of lncRNA H19 in 24 pairs of BC tissues and 20 pairs of BC plasma. The differentially expressed of plasma H19 was further validated in another 102 BC patients and 96 healthy controls. The potential correlations between plasma H19 levels and clinicopathological characteristics were analyzed. Receiver operating characteristic (ROC) curve was performed to evaluate the diagnostic values of plasma H19 between 30 early stage BC patients and 30 healthy controls. 24 paired pre- and postoperative plasma samples were detected to assess the tumor monitoring values. RESULTS: The results revealed that the expression of H19 was significantly increased in BC tissues and plasma compared with healthy controls (P< 0.05), and plasma H19 levels were significantly correlated with estrogen receptor (ER) (P= 0.008), progesterone receptor (PR) (P= 0.025), c-erbB-2 (P= 0.043) and lymph node metastasis (P= 0.006). The area under the ROC curve (AUC) of plasma H19 was 0.81(sensitivity, 56.7%; specificity, 86.7%; P< 0.0001), which was higher than the values of carcinoembryonic antigen (CEA) and carbohydrate antigen 153 (CA153). Furthermore, plasma H19 levels were significantly decreased in postoperative samples than preoperative samples (P= 0.0006). CONCLUSION: Plasma H19 may serve as a potential biomarker for BC early screening and prognosis monitor. Show more

Keywords: Breast cancer, lncRNA H19, plasma biomarker

DOI: 10.3233/CBM-160630

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 187-194, 2016

Authors: Gong, Long | Xu, Ying | Hu, Ya-Qin | Ding, Qiu-Ju | Yi, Chun-Hua | Huang, Wei | Zhou, Ming

Article Type: Research Article

Abstract: OBJECTIVES: The study explored the association between rs10069690C/T and rs2736100G/T of human telomerase reverse transcriptase (hTERT ) gene, and the prognosis of thyroid cancer. METHODS: The study had 452 thyroid cancer patients recruited as case group who hospitalized in Jingzhou Central Hospital from January 2001 to June 2004 and 452 healthy people recruited as control group at the same area. The hTERT gene polymorphisms at rs10069690 C/T and rs2736100 G/T were tested by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The association between patients' life quality and hTERT gene polymorphisms six months after …surgery was evaluated based on the Cancer patients' quality of life index rating scale. RESULTS: There were statistical differences in genotype and allele frequencies of rs10069690 C/T between the case group and control group (both P < 0.05). An association between rs10069690C/T polymorphism and an increased risk of thyroid cancer was shown by logistic regression analysis (CT vs. CC, OR = 1.333, 95%CI = 1.006-1.766, P = 0.045; TT vs. CC, OR = 1.910, 95%CI = 1.084-3.367, P = 0.023; CT + TT vs. CC, OR = 2.246, 95%CI = 1.078-1.840, P = 0.006; T vs. C, OR = 1.376, 95%CI = 1.104-1.715, P = 0.004). Genotype frequency of rs2736100G/T between the two groups had no statistical differences (P > 0.05). After stratification according to age, T stage, tumor size and tumor node metastasis (TNM) stage, the distribution frequencies of CC genotype and CT + TT genotype of rs10069690C/T showed significant difference (P < 0.05). The life quality of patients with CC genotype was better than that of patients with CT $+$ TT genotype. The results of Cox regression model multifactor analysis showed that age, T stage, tumor size and rs10069690C/T were independent risk factors of thyroid cancer prognosis. CONCLUSIONS: hTERT gene polymorphism at rs10069690C/T is associated with the risk and prognosis of thyroid cancer, but hTERT gene polymorphism at rs2736100G/T is not. Show more

Keywords: Human telomerase reverse transcriptase (hTERT), thyroid cancer, gene polymorphism, prognosis, rs10069690C/T, rs2736100G/T, life quality evaluation

DOI: 10.3233/CBM-160631

Citation: Cancer Biomarkers, vol. 17, no. 2, pp. 195-204, 2016