Kyu Yeoun Won

Clinicopathologic correlation of beclin-1 and bcl-2 expression in human breast cancer

The human beclin-1 gene, located on chromosome 17q21, has been identified as the mammalian orthologue of Atg6 (autophagy-related gene) and may be a haploinsufficient tumor suppressor gene. The function and expression of beclin-1 in human breast cancer are largely unknown. We investigated the expression of beclin-1 and bcl-2 in human breast cancer. Tissue samples from 125 cases of invasive breast cancer were used for the present study. Immunohistochemical staining for beclin-1 and bcl-2 was evaluated using tissue microarray, then the 2 proteins were correlated with clinicopathologic parameters. Positive beclin-1 expression and bcl-2 expression in breast cancer tissue were observed in 53 cases (42.4%) and 48 cases (38.4%), respectively. Beclin-1 expression was inversely correlated with bcl-2 expression in breast cancer tissue (P = .035). Beclin-1 expression significantly correlated with nuclear pleomorphism and mitotic count. Bcl-2 expression in breast cancer tissue significantly correlated with histologic grade, tubule formation, nuclear pleomorphism, mitotic count, estrogen receptor, and distant metastasis. Our results suggest that beclin-1 might play a role in the inhibition of the development of breast cancer and that inhibition might be due to an interaction with bcl-2 protein.

Regulatory role of p53 in cancer metabolism via SCO2 and TIGAR in human breast cancer

Cancer cells show a higher rate of anaerobic respiration than normal cells. The exact mechanisms for this higher glycolysis rate in cancer cells remain to be elucidated. The results of recent studies have indicated that p53, the most commonly mutated tumor suppressor gene, may have important functions in the regulation of energy-generating metabolic pathways that switch from oxidative phosphorylation to glycolysis via the synthesis of cytochrome c oxidase 2 (SCO2), p53-transactivated TP53-induced glycolysis (TIGAR), and apoptosis regulator. We evaluated the expression of p53, SCO2, TIGAR, and COX in 113 cases of invasive breast cancer using immunohistochemistry. A high expression of p53, SCO2, TIGAR, and COX was noted in 27.5% (31 cases), 84.1% (95 cases), 74.3% (84 cases), and 73.4% (83 cases) of the breast tumors, respectively. A high p53 expression was significantly associated with low expression levels of SCO2 (P = .008), COX (P < .0001), and TIGAR (P = .007). On the survival analysis, the low SCO2-expressing breast cancer patients showed a significantly poorer prognosis than that of the high SCO2-expressing breast cancer patients (P = .0078). These results suggest that p53 can modulate the metabolic pathways via the proteins SCO2 and TIGAR in human breast cancer.

Prognostic implication of immunohistochemical Runx2 expression in osteosarcoma.

Aims and background Osteosarcoma is the most common primary bone malignancy. Many genetic markers have proven prognostic value in osteosarcoma and studies are under way to determine their potential application as specific therapeutic targets. Runx2, Indian hedgehog (IHH), and Sox9 are proteins that play major roles in bone formation and tumorigenesis. We studied the protein expression of Runx2, IHH, and Sox9 in osteosarcoma and correlated their expression with clinicopathological variables. We also studied the prognostic value of the expression of these three genes in osteosarcoma. Methods and study design We produced 48 formalin-fixed, paraffin-embedded tissue microarrays containing osteosarcoma tissue cores for immunohistochemical staining of Runx2, IHH and Sox9. We evaluated the expression of each gene by immunohistochemical staining and analyzed the relationship between expression and clinicopathological parameters. Results High expression of Runx2 was significantly related to metastasis (P = 0.015). High expression of Runx2 indicated a trend toward a poor survival rate (P = 0.056). High expression of IHH and Sox9 were not related to any clinicopathological parameters. Conclusions High expression of Runx2 was significantly related to tumor metastasis in osteosarcoma. Our results suggest that overexpression of Runx2 might be a useful prognostic marker in osteosarcoma cases.

MicroRNA-199b-5p is involved in the Notch signaling pathway in osteosarcoma

MicroRNAs (miRNAs) play important roles in the development, differentiation, and function of different cell types and in the pathogenesis of various human diseases. miRNAs are differentially expressed in normal and cancer cells. The investigation of miRNA expression between healthy subjects and patients with osteosarcoma is crucial for future clinical trials. We performed miRNA microarray analysis on 8 formalin-fixed, paraffin-embedded osteosarcoma tissue samples. We confirmed the results of the microarray analysis using reverse transcription polymerase chain reaction. miRNA profiling of osteosarcoma tissue samples showed that expression of 10 miRNAs had increased 10-fold compared with normal controls. Among the 10 miRNAs, 3 miRNAs (miR-199b-5p, miR-338-3p, and miR-891a) were confirmed to have been up-regulated by reverse transcription polymerase chain reaction. After transfection of 4 osteosarcoma cell lines with miR-199b-5p inhibitor, the expression of Notch pathway components in the transfected cell lines was changed. These results revealed that miR-199b-5p plays a role in Notch signaling in osteosarcoma. Recently, the inhibition of Notch and HES1 signaling has been suggested as a potential therapeutic strategy to prevent metastasis in human osteosarcoma. Taken together with our results, we suggest that miR-199b-5p inhibitor may also be a therapeutic option for osteosarcoma.

Decreased Beclin-1 expression is correlated with the growth of the primary tumor in patients with squamous cell carcinoma and adenocarcinoma of the lung

Beclin-1 is a Bcl-2-interacting protein, and it may delay cell cycle progression and induce autophagy. The function and expression of Beclin-1 and Bcl-2 in squamous cell carcinoma and adenocarcinoma of the lung remain largely unknown. Herein, we investigated the expression of Beclin-1 and Bcl-2 in squamous cell carcinoma and adenocarcinoma of the lung. Tissue samples from 262 cases were used in this study. Immunohistochemical staining for Beclin-1 and Bcl-2 were conducted using a tissue microarray. In squamous cell carcinoma, Beclin-1 expression was strongly positive in 48 (28.6%) of 168 samples, it was moderately positive in 42 (25.0%) of 168 samples, and it was negative or weakly positive in 78 (46.4%) of 168 samples. In adenocarcinoma, Beclin-1 expression was strongly positive in 26 (27.7%) of 94 samples, it was moderately positive in 27 (28.7%) of 94 samples, and it was negative or weakly positive in 41 (43.6%) of 94 samples. Beclin-1 expression was inversely correlated with the tumor size and the primary tumor stage (pT) in both types of tumor. Especially, the TNM stage of adenocarcinoma was inversely correlated with Beclin-1 expression. Our results suggest that a progressively reduced Beclin-1 expression is correlated with the primary tumor growth of squamous cell carcinoma and adenocarcinoma of the lung.

RANK signalling in bone lesions with osteoclast-like giant cells.

Aims: The interactions between the receptor activator of NF-κB (RANK), its ligand (RANKL), and the decoy receptor for RANKL, osteoprotegerin (OPG), play a pivotal role in promoting osteoclast differentiation and activation leading to bone resorption. Giant cell tumours, chondroblastomas, and aneurysmal bone cysts harbour osteolytic lesions containing osteoclast-like giant cells. We investigated the characteristics of the RANKL signalling pathway in each of these bone lesions. Methods: We evaluated 44 cases of giant cell tumour, 12 cases of chondroblastoma, six cases of aneurysmal bone cyst, and five cases of metastatic giant cell tumour (including paired primary giant cell tumours). We assessed RANK, RANKL, and OPG expression in chondroblastomas, giant cell tumours, and aneurysmal bone cysts using immunohistochemical methods. Results: Our findings revealed that RANK, RANKL, and OPG expression differed significantly among disease types. Giant cells of chondroblastomas showed significantly higher RANK expression than the giant cells of giant cell tumours and aneurysmal bone cysts; similarly, stromal cells of chondroblastomas showed significantly higher OPG expression than the stromal cells of giant cell tumours and aneurysmal bone cysts. Furthermore, giant cells of giant cell tumours expressed significantly more RANK than the giant cells of aneurysmal bone cysts. Conclusions: The expression of RANK, RANKL, and OPG in osteoclast-like giant cells differs significantly by disease; OPG expression differs significantly between giant cell tumours and chondroblastomas.

Reduced expression of Raf-1 kinase inhibitory protein predicts regional lymph node metastasis and shorter survival in esophageal squamous cell carcinoma

Raf-1 kinase inhibitory protein (RKIP), a suppressor of metastasis, is associated inversely with the progression and metastasis of human malignancies. The present study evaluated relationships between RKIP expression and metastatic potential, clinicopathological characteristics and patient outcome in esophageal squamous cell carcinoma (ESCC). We examined tissue specimens from 138 patients with thoracic ESCC. Using immunohistochemistry, RKIP expression was detected in ESCC in situ, primary ESCC and nodal metastatic ESCC. RKIP expression was reduced in 28.9% (13/45) of ESCC in situ, in 50.0% (69/138) of primary ESCC and in 71.4% (65/91) of nodal metastatic ESCC. These levels of RKIP down-regulation differed significantly. RKIP expression was associated inversely with histological grade (P=0.008), pathological T stage (P=0.044), lymphatic invasion (P=0.019), regional lymph node metastasis (LNM; P=0.002) and stage (P=0.041). Pathological T stage (P=0.001), lymphatic invasion (P<0.001) and reduced RKIP expression (P=0.039) were independent predictors of regional LNM in ESCC. In addition, the postoperative survival of patients with RKIP-reduced ESCC was significantly shorter than for patients with RKIP-positive ESCC (P=0.004). Reduced RKIP expression in ESCC correlated with advanced disease, regional LNM and poor prognosis. RKIP expression may serve as a novel clinical biomarker in patients with ESCC.

Polarized CD163+ tumor-associated macrophages are associated with increased angiogenesis and CXCL12 expression in gastric cancer.

PURPOSE Tumor-associated macrophages (TAMs) play a significant role in tumor progression and angiogenesis. However, the prognostic value of TAMs in different histologic locations of gastric cancer (GC) is still unknown. We evaluated the distribution of TAMs in different histologic locations to investigate its importance in predicting prognosis and the relationship with angiogenesis and CXCL12 expression in GC. METHODS AND MATERIALS The distribution of TAMs and microvessel density (MVD) in 113 GC samples were evaluated by immunohistochemical staining of CD163 and CD105, respectively. The extent of TAM distribution in the tumor was categorized into three groups: infiltrated TAMs in the tumor nest (TN), tumor stroma (TS) and invasive tumor margin (TM). The expression of CXCL12 in GC were evaluated by immunohistochemical staining of tissues from 88 GC samples. RESULTS The increased CD163+ TAMs in TS and TM were closely correlated with tumor size, depth of invasion, TNM stage, lymph node metastasis, and lymphovascular invasion. TAMs in TN was not related with any clinicopathologic characteristics except histologic differentiation. The high infiltration of CD163+ TAMs in TS and TM were significantly correlated with poor overall survival. Regardless of location, CD163+ TAMs were significantly correlated with increased MVD. CXCL12 expression was significantly associated with increased CD163+ TAMs in TS and TM. CONCLUSIONS TAMs in different histologic locations in GC were related to distinct aspects of tumor progression. CD163+ TAMs in TS and TM are associated with tumor progression and CXCL12 expression in GC. TAMs may be involved in tumor progression through the angiogenesis.

Expression of BDNF, TrkB, and p53 in early-stage squamous cell carcinoma of the uterine cervix.

Aims: TrkB and BDNF play a critical role in cancer invasion and metastasis. We investigated the potential role of these proteins, together with p53, in squamous cell carcinoma (SCC) of the uterine cervix by focusing on the clinicopathological correlation and outcome. Methods: We performed RT-PCR analysis of TrkB and BDNF mRNA expression in cervical cancer cell lines, as well as tumour and normal tissue of the cervix. We analysed TrkB, BDNF, and p53 protein expression using immunohistochemistry in 80 patients with cervical SCC. Results: The expression of BDNF and TrkB mRNA was higher in human cervical cancer cell lines and SCC tissues than in normal tissues. Positive TrkB expression was significantly correlated with low FIGO stage (p = 0.032), low T stage (p = 0.027), less than 1/2 stromal invasion of the cervix (p = 0.018), 4 cm or smaller for the greatest dimension of the tumour (p = 0.015) and absent lymphatic invasion (p = 0.032). Positive BDNF expression was significantly related to low FIGO stage (p = 0.001), less than 1/2 stromal invasion of the cervix (p = 0.040), absent lymphatic invasion (p = 0.026), and absent lymph node metastasis (p = 0.003). In contrast, p53 expression was significantly related to advanced T stage (p = 0.001), greater than 4 cm for the greatest dimension of the tumour (p = 0.002), more than 1/2 stromal invasion of the cervix (p = 0.005), and poor survival (p = 0.0218). Conclusions: TrkB and BDNF expression might play a significant role in the early events in tumorigenesis of cervical SCC. However, p53 is involved late in the process of tumour progression and therefore is predictive of a poor prognosis in SCC.

Absolute monocyte and lymphocyte count prognostic score for patients with gastric cancer

AIM To measure the prognostic significance of absolute monocyte count/absolute lymphocyte count prognostic score (AMLPS) in patients with gastric cancer. METHODS We retrospectively examined the combination of absolute monocyte count (AMC) and absolute lymphocyte count (ALC) as prognostic variables in a cohort of 299 gastric cancer patients who underwent surgical resection between 2006 and 2013 and were followed at a single institution. Both AMC and ALC were dichotomized into two groups using cut-off points determined by receiving operator characteristic curve analysis. An AMLPS was generated, which stratified patients into three risk groups: low risk (both low AMC and high ALC), intermediate risk (either high AMC or low ALC), and high risk (both high AMC and low ALC). The primary objective of the study was to validate the impact of AMLPS on both disease-free survival (DFS) and overall survival (OS), and the second objective was to assess the AMLPS as an independent prognostic factor for survival in comparison with known prognostic factors. RESULTS Using data from the entire cohort, the most discriminative cut-off values of AMC and ALC selected on the receiver operating characteristic curve were 672.4/μL and 1734/μL for DFS and OS. AMLPS risk groups included 158 (52.8%) patients in the low-risk, 128 (42.8%) in the intermediate-risk, and 13 (4.3%) in the high-risk group. With a median follow-up of 37.2 mo (range: 1.7-91.4 mo), five-year DFS rates in the low-, intermediate-, and high-risk groups were 83.4%, 78.7%, and 19.8%, respectively. And five-year OS rates in the low-, intermediate-, and high-risk groups were 89.3%, 81.1%, and 14.4%, respectively. On multivariate analysis performed with patient- and tumor-related factors, we identified AMLPS, age, and pathologic tumor-node-metastasis stage as the most valuable prognostic factors impacting DFS and OS. CONCLUSION AMLPS identified patients with a poor DFS and OS, and it was independent of age, pathologic stage, and various inflammatory markers.