Atypical Hyperplasia Tissues Research Articles

Expression of LATS1 in head and neck squamous cell carcinoma and its effect on tumor cell proliferation and invasion.

Nowadays, the incidence and mortality of head and neck tumors are gradually increasing. Head and neck malignant tumors (such as laryngeal cancer, hypopharyngeal cancer, oral cancer, nasopharyngeal cancer, oropharyngeal cancer, and other head and neck malignancies) are more common among systemic tumors. The most common pathological head and neck tumor type is squamous cell carcinoma, accounting for about 90%. In this study, immunohistochemical methods were used to collect the normal squamous epithelial tissues of the head and neck, atypical hyperplasia tissues, and head and neck squamous cell carcinomas on a tissue chip for detection. The recombinant LATS1 overexpression plasmid was prepared and transferred into B88 cells. Western blotting, MTT, and Transwell chamber methods were used to detect the effects of LATS1 proliferation, migration, and B88 cell overexpression. The experimental results showed that in head and neck squamous cell carcinoma, the expression of LATS1 protein decreased from 59.3% to 11.3%. At the same time, this protein inhibited the proliferation, migration, and invasion of head and neck squamous epithelial cells and also inhibited epithelium- Interstitial transformation exerts its tumor suppressor effect, indicating that LATS1 may play a tumor suppressor effect as a tumor suppressor gene. An in-depth study of the role and mechanism of LATS1 protein in the occurrence of head and neck squamous cell carcinoma may provide new opportunities for the treatment of head and neck squamous cell carcinoma in the future.

Expression and clinical significance of CXCL17 and GPR35 in endometrial carcinoma.

Endometrial carcinoma is one of the most common gynecologic malignancies. CXCL17-CXCR8 (GPR35) axis is reported to play an indispensability role in tumors. Our purpose is to screen possible prognostic and immune-related factors in endometrial carcinoma by detecting the mRNA and protein expression of CXCL17 and CXCR8. We use the qRT-PCR method to test the mRNA expression of CXCL17 and CXCR8 in 35 pairs of endometrial carcinoma and adjacent tissue. The protein expression of CXCL17 and CXCR8 in 30 cases of normal proliferative endometrium, 30 cases of endometrial atypical hyperplasia and 50 cases of endometrial carcinoma was detected by tissue microarray immunohistochemistry. There was no significant difference in the positive expression rate between endometrial adenocarcinoma tissue and endometrial atypical hyperplasia tissue (P > 0.05). But significantly better than normal proliferative tissue (P < 0.001). Correlation analysis of CXCR8 and CXCL17 in endometrial carcinoma showed a positive correlation (r = 0.9123, P < 0.0001). For patients with endometrial cancer, the overall survival (OS) of patients with high CXCL17 expression was significantly higher than that low CXCL17 expression (log-rank test, P < 0.0001), whereas CXCR8 had no statistical significance. But the expression of CXCR8 is an independent prognostic factor of OS in endometrial carcinoma patients. Our study showed that CXCL17 and CXCR8 may be involved in the occurrence and development of endometrial cancer. High expression of CXCL17 may be used as a biomarker for predicting survival. Because CXCL17 and CXCL18 are related to lymphocytes and immune regulation, they are expected to become potential targets for immunotherapy.

Spectroscopic evaluation of carcinogenesis in endometrial cancer

Carcinogenesis is a multifaceted process of cancer formation. The transformation of normal cells into cancerous ones may be difficult to determine at a very early stage. Therefore, methods enabling identification of initial changes caused by cancer require novel approaches. Although physical spectroscopic methods such as FT-Raman and Fourier Transform InfraRed (FTIR) are used to detect chemical changes in cancer tissues, their potential has not been investigated with respect to carcinogenesis. The study aimed to evaluate the usefulness of FT-Raman and FTIR spectroscopy as diagnostic methods of endometrial cancer carcinogenesis. The results indicated development of endometrial cancer was accompanied with chemical changes in nucleic acid, amide I and lipids in Raman spectra. FTIR spectra showed that tissues with development of carcinogenesis were characterized by changes in carbohydrates and amides vibrations. Principal component analysis and hierarchical cluster analysis of Raman spectra demonstrated similarity of tissues with cancer cells and lesions considered precursor of cancer (complex atypical hyperplasia), however they differed from the control samples. Pearson correlation test showed correlation between cancer and complex atypical hyperplasia tissues and between non-cancerous tissue samples. The results of the study indicate that Raman spectroscopy is more effective in assessing the development of carcinogenesis in endometrial cancer than FTIR.

KRAS, YWHAE, SP1 and MSRA as biomarkers in endometrial cancer.

BackgroundWe demonstrated that drinking hydrogen-rich water (HRW) inhibits endometrial tumor growth in our previous work. This research is to identify differentially expressed proteins (DEPs) between HRW and purified water groups in a xenograft mouse model of endometrial cancer (EC).MethodsSamples were analyzed using tandem mass tags (TMTs) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). DEPs were identified using bioinformatics to determine potential molecular functions and immunohistochemical (IHC) staining.ResultsIn total, 11 DEPs were identified in the HRW group relative to the control. The up-regulated proteins included Gatad1, Ttyh3, Nek4, Dyrk2, and Gimap1, while the down-regulated proteins included SP1, Msl1, Plekha7, Dtwd2, MSRA, and KRAS. Gene Ontology (GO) annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were associated with the binding region, biological regulation, endocrine resistance, estrogen signaling, choline metabolism in cancer and human cytomegalovirus infection. Furthermore, network analysis indicated that KRAS and MSRA interact with YWHAE. KRAS, YWHAE and SP1 were strongly expressed, while MSRA was weak expressed in atypical hyperplasia and EC tissue as well as in HRW group in xenograft tumor tissue.ConclusionsKRAS, YWHAE, SP1 and MSRA might be regarded as focused biomarkers to assess the prognosis of EC.

Simultaneous FTIR and Raman Spectroscopy in Endometrial Atypical Hyperplasia and Cancer.

Currently, endometrial carcinoma (EC) is the most common genital cancer in high-income countries. Some types of endometrial hyperplasia (EH) may be progressing to this malignancy. The diagnosis of EC and EH is based on time consuming histopathology evaluation, which is subjective and causes discrepancies in reassessment. Therefore, there is a need to create methods of objective evaluation allowing the diagnosis of early changes. The study aimed to simultaneously asses Fourier Transform Infrared (FTIR) and Raman spectroscopy combined with multidimensional analysis to identify the tissues of endometrial cancer, atypical hyperplasia and the normal control group, and differentiate them. The results of FTIR and Raman spectroscopy revealed quantitative and qualitative changes in the nucleic acid and protein in the groups of cancer and atypical hyperplasia, in comparison with the control group. Changes in the lipid region were also observed in Raman spectra. Pearson correlation coefficient demonstrated a statistically significant correlation between Raman spectra for the cancer and atypical hyperplasia groups (0.747, p < 0.05) and for atypical hyperplasia and the controls (0.507, p < 0.05), while FTIR spectra demonstrated a statistically significant positive correlation for the same group as in Raman data and for the control and cancer groups (0.966, p < 0.05). To summarize, the method of spectroscopy enables differentiation of atypical hyperplasia and endometrial cancer tissues from the physiological endometrial tissue.

Decreased Nek9 expression correlates with aggressive behaviour and predicts unfavourable prognosis in breast cancer

As a new member of Neks family, Nek9 regulates spindle assembly and controls chromosome alignment and centrosome separation. In the current study we aimed to investigate the expression of Nek9 in breast cancer and its clinical significance. We evaluated the expression of Nek9 in invasive ductal carcinoma (IDC, n=316), ductal carcinoma in situ (DCIS), usual ductal hyperplasia, atypical ductal hyperplasia, fibroadenoma and normal breast tissues using immunohistochemistry. The results revealed significantly reduced Nek9 in IDCs (41.8%) compared to benign breast lesions. Moreover, gradually reduced Nek9 was found from DCIS to invasive carcinoma and metastatic tumour within the same tumours. The decrease in Nek9 expression was associated with larger tumour size (p=0.0087), high grade (p<0.0001) and high Ki-67 index (p<0.0020). TCGA and GEO datasets analysis revealed low level of Nek9 mRNA was more frequent in triple negative breast cancers, and associated with poor overall survival and distant metastasis-free survival. These findings suggest an important role of Nek9 in the progression of breast cancer, and aberrantly expressed Nek9 correlates with more aggressive clinicopathological variables and predicts poor clinical prognosis. Nek9 may serve as a potential predictive factor for patients with breast cancer.

Estrogen/estrogen receptor promotes the proliferation of endometrial carcinoma cells by enhancing hMOF expression.

BackgroundThis study aims to analyse the expression of human MOF in endometrial carcinoma cells and its relationship with estrogen and estrogen receptor and to investigate the effect of estrogen–human MOF on the malignant biological behaviours of endometrial carcinoma cells.MethodsThe expression of human MOF was detected in different endometrial tissues by immunohistochemistry. The effects of human MOF, human MOF combined with estrogen stimulation and estrogen plus anti-human MOF antibody blocking on the proliferation of endometrial carcinoma cells were evaluated by western blotting, real-time polymerase chain reaction, cell proliferation assay and cell cycle distribution. Bioinformatics was used to identify the correlations of human MOF and estrogen and involved pathways.ResultsThe expression levels of human MOF in endometrial carcinoma tissues were significantly higher than that in atypical hyperplasia and normal endometrial tissues. High expression of human MOF was associated with late-stage cancer, lymph node metastasis and short survival time, and it was also an independent prognostic risk factor for endometrial carcinoma. After human MOF knockdown, the proliferation, migration and invasive capacity of Ishikawa cells decreased and cell apoptosis increased. After stimulation with estrogen, the PI3K/Akt and Ras–Raf–MEK–ERK signalling pathways were activated, and the expression of the human MOF protein was increased. human MOF (KAT8) expression showed a positive correlation with ESR1 expression, and KAT8-associated genes were enriched in the cell cycle pathways and splicing pathways.ConclusionHuman MOF was highly expressed in endometrial carcinoma and associated with proliferation. Estrogen/estrogen receptor enhanced human MOF expression; promoted the proliferation, migration and invasion of Ishikawa cells; and inhibited cell apoptosis by activating the PI3K/Akt and Ras–Raf–MEK–ERK signalling pathways.

Proteomic analysis in endometrial cancer and endometrial hyperplasia tissues by 2D-DIGE technique

ObjectiveTo compare the protein expression of complex atypical endometrial hyperplasia, endometrial carcinoma and healthy endometrial tissues, and by this way, to identify proteins that can be used for diagnosis, prognosis and therapeutic targets. MethodsHistopathological examination of the D&C material had reported “benign endometrial changes”, “complex atypical endometrial hyperplasia” and “endometrioid adenocarcinoma” and 30 patients ,who underwent surgery with these diagnosis, were studied.Protein profiles of the study groups were detected using 2D-DIGE technique and compared to the control group. Protein spots which showing different expression, were defined by MALDI TOF/TOF-MS method. ResultsIn the present study, significant elevations were observed in the levels of K2C8, UAP56, ENOA, ACTB, GRP78, GSTP1, PSME1, CALR, PPIA, PDIA3 and IDHc proteins when comparisons were made among the cancer cases and the healthy and complex atypical hyperplasia cases. We determined that the induction of CALR activity may be a factor that progresses apoptosis, thus, may be a hope for postoperative new chemotherapy treatment methods. Moreover, when the expressions of the CAH1 and PPIB proteins are compared to complex atypical hyperplasia and endometrial adenocarcinoma stages, we determined that the CAH1 and PPIB levels increased in more advanced stages. Among these indicators, the proteins that had the closest relation to advanced stage cancer were determined as K2C8, UAP56 and GRP78. ConclusionWe think that it would be useful to determine the diagnosis, prediction of prognosis and identifying therapeutic targets of the highlighted proteins of our study that are K2C8, UAP56, GRP78 and CALR in endometrial cancer.

Expressions of growth factor receptor binding protein 7 and human epidermal growth factor receptor 2 in endometrial carcinoma and their significances

Objective To investigate the expressions of growth factor receptor binding protein 7(GRB7) and human epidermal growth factor receptor 2 (HER2) in endometrial carcinoma and their clinical significances. Methods Immunohistochemistry was used to detect the expressions of GRB7 and HER2 in 52 endometrial carcinoma tissues, 38 endometrial atypical hyperplasia tissues and 30 normal endometrial tissues collected from patients in the Sixth People's Hospital of Shanghai Jiao Tong University from May 2015 to September 2017. The relationship between the expressions of GRB7 and HER2 and clinicopathological features of endometrial carcinoma patients was analyzed. Results The positive expression rates of GRB7 in endometrial carcinoma, endometrial atypical hyperplasia and normal endometrial tissues were 71.2% (37/52), 39.5% (15/38), and 13.3% (2/30), respectively, and the difference was statistically significant (χ2 = 35.14, P < 0.05). The positive expression rates of HER2 in endometrial carcinoma, endometrial atypical hyperplasia and normal endometrial tissues were 67.3% (35/52), 34.2% (13/38), and 10.0% (3/30), respectively, and the difference was statistically significant (χ2 = 29.360, P < 0.05). The positive expression rates of GRB7 in endometrial carcinoma patients with different clinical stages, histological grades and pathological types were statistically significant (all P < 0.05). The positive expression rates of HER2 in endometrial carcinoma patients with different clinical stages, histological grades, lymph nodes metastasis and myometrial invasion were statistically significant (all P < 0.05). The expression of GRB7 in endometrial carcinoma was positively correlated with the expression of HER2(r = 0.375, P < 0.05). Conclusion The high expressions of GRB7 and HER2 in endometrial carcinoma are closely related to the malignant degree of endometrial carcinoma. Key words: Endometrial neoplasms; Growth factor receptor binding protein 7; Human epidermal growth factor receptor 2; Neoplasm staging

Effects of long noncoding RNA (linc-VLDLR) existing in extracellular vesicles on the occurrence and multidrug resistance of esophageal cancer cells

ObjectiveTo investigate the relationship between the expression of linc-VLDLR in extracellular vesicles (EVs) and esophageal carcinomas development and drug resistance. MethodsThe expression of linc-VLDLR and ABCG2 mRNA in 60 cases of esophageal carcinoma tissue, para-carcinoma tissue and the normal esophagus tissue were detected using Fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR). Fifty percent inhibiting concentration (IC50) of adriamycin (ADM) to Eca109 cells was detected by MTT assay, after the treatment of different concentrations of adriamycin (ADM) on esophageal squamous cell carcinoma Eca109 cell line for 24 h. EVs were extracted from culture medium after the treatment of three concentrations of ADM (setting based on the IC50) on Eca109 cells for 24 h. Linc-VLDLR expression in EVs was detected by qRT-PCR. After the treatment of the extracted EVs on virgin Eca109 cells for 48 h, then intervening these cells for 24 h by different concentrations of ADM, the new values of IC50 were detected by MTT assay. Cell cycle, cell apoptosis and ABCG2 protein expression of these Eca109 cells were detected by flow cytometry (FCM). Linc-VLDLR and ABCG2 mRNA expression in these Eca109 cells were detected by qRT-PCR. ResultsExpression of linc-VLDLR and ABCG2 mRNA in esophageal squamous cell carcinoma tissue were significantly higher than that in esophageal atypical hyperplasia and normal esophagus tissue, P < 0.01. After the treatment of ADM on Eca109 cells for 24 h, IC50 of Eca109 cells was detected as (0.44 ± 0.02) μg/mL, thus ADM concentrations of 0, 0.2, 0.4 and 0.8 μg/mL were selected to accomplish the following parts of this study. After four groups of Eca109 cells were treated by ADM in different concentrations separately, extracted EVs from the supernatant of all four groups, then labeling these four groups as EVs1, 2, 3 and 4. Linc-VLDLR expression in EVs4 was significantly higher than that in EVs1-3, P < 0.01. After the treatment of EVs1-4 on virgin Eca109 cells for 48 h, new values of IC50 of Eca109 to ADM were detected by MTT. It was found that the IC50 value of group EVs4 was significantly higher than that of other groups, P < 0.05. Flow cytometry results showed that the proliferation index of Eca109 cells in EVs4 was significantly higher than that in EVs1-3 and control groups, P < 0.01. Whereas, there was an obviously downward trend in the apoptosis rate of EVs4, compared to other three groups, P < 0.01. Linc-VLDLR and ABCG2 mRNA and protein expression level in Eca109 cells of EVs4 group were significantly higher than that of EVs1-3 and control groups, P < 0.05. ConclusionsHigh expression of Linc-VLDLR and ABCG2 gene in esophageal cancer cells affected the formation of esophageal cancer drug resistance. EVs released by drug-resistant cells were proved that they could upregulate the expression of ABCG2 in esophageal cancer cells and thus regulate the drug resistance of esophageal cancer cells, which was related to the linc-VLDLR carried by EVs.

BKCa participates in E2 inducing endometrial adenocarcinoma by activating MEK/ERK pathway

BackgroundThe large-conductance, voltage-gated, calcium (Ca (2+))-activated potassium channel (BKCa) plays an important role in regulating Ca (2+) signaling and cell physiological function, and is aberrantly expressed in some types of cancers. The present study focuses on identifying the oncogenic potential and clinical significance of BKCa in endometrial adenocarcinoma, as well as exploring the mechanistic relevance by 17β -estradiol (E2) inducing aberrant activation of MEK1/2 and ERK1/2 via BKCa.MethodsThe expression of BKCa, ERK1/2 and p-ERK1/2 were examined by immunohistochemical staining in 263 cases, including 185 primary types I endometrial cancer tissues, 38 atypical endometrial hyperplasia tissues and 40 normal endometrium tissues. Cell growth, cycle, apoptosis rate, migration and invasion was separately tested in Ishikawa cells using siRNA-BKCa and/or E2 treatment, as well as the expression of these interested proteins by western blot analysis.ResultsWe showed that expression of BKCa is significantly elevated in 185 types I endometrial adenocarcinoma tissues compared to those of the normal endometrium and atypical endometrial hyperplasia tissues. Furthermore, in vitro observations revealed that down-regulation of BKCa expression inhibited cell growth by both enhancing apoptosis and blocking G1/S transition, suppressed cell migration and invasion in Ishakiwa cells, and decreased the expression of p-MEK1/2 and p-ERK1/2. Additionally, RNAi-mediated knockdown of BKCa attenuated the increased cellular growth and invasion, as well as the elevated expression of p-MEK1/2 and p-ERK1/2 proteins, induced by E2 stimulation. More importantly, the aberrant expression of BKCa and p-ERK1/2 were closely related with poor prognostic factors in type I endometrial cancer, and up-regulated expression of p-ERK1/2 was significantly associated with shorter disease-free survival (DFS) and overall survival (OS) and was an independent prognostic factor in type I endometrial cancer patients.ConclusionOur results demonstrated that BKCa and the key downstream effectors p-ERK1/2 could be involved in important signaling pathways in initiation and development of endometrial adenocarcinoma and may provide a new therapeutic approach for women with endometrial cancer.

Expression of ELF5 in endometrial carcinoma tissues and its clinical significance.

Expression of E74-like factor 5 (ELF5) in endometrial carcinoma tissues and its clinical significance were investigated. Eighty-four endometrial carcinoma tissues, 30 cases of atypical hyperplasia of endometrium and 30 cases of normal endometrial tissues were selected. Immunohistochemical method was utilized to detect the expression of ELF5 in different endometrial tissues. Moreover, its correlation with clinical pathological indexes of patients with endometrial carcinoma was analyzed. The postoperative follow-up was conducted in all the patients with endometrial carcinoma until June 30th, 2017. The Kaplan-Meier method was used for survival analysis so as to analyze the association of ELF5 expression level with clinical pathological indexes; Cox's proportional hazards regression model was utilized for univariate and multivariate analyses to screen independent risk factors for prognosis of endometrial carcinoma. In normal endometrial tissues, atypical hyperplasia and endometrial carcinoma tissues, the positive expression rates of ELF5 showed a decreased tendency (P=0.016). The positive expression rate of ELF5 in endometrial carcinoma tissues was lower in comparison to normal endometrial tissues (P=0.016). The expression of ELF5 was in accordance with the International Federation of Gynecology and Obstetrics (FIGO) staging of endometrial carcinoma (P<0.05), pathological grading (P<0.05), pathological typing (P=0.001), state of lymph node metastasis (P<0.05) and depth of myometrial invasion (P<0.05). Kaplan-Meier method for survival analysis showed that the average survival time of patients with negative ELF5 expression was shorter in comparison to the patients with positive expression (P=0.004). FIGO staging (P=0.004), pathological grading (P=0.048), depth of myometrial invasion (P=0.024) and lymph node metastasis (P=0.020) were related to the prognosis of patients with endometrial carcinoma, The univariate Cox's regression model analysis indicated that FIGO staging (P=0.010), pathological grading (P=0.040), depth of myometrial invasion (P=0.037), lymph node metastasis (P=0.029) and ELF5 (P=0.010) were associated with the prognosis of patients with endometrial carcinoma. Further, multivariate analysis revealed that ELF5 was an independent risk factor for prognosis of patients with endometrial carcinoma (P=0.035). The expression of ELF5 has a correlation with the occurrence, development and prognosis of endometrial carcinoma.

Identification of differentially expressed proteins in the gastric mucosal atypical hyperplasia tissue microenvironment

In the present study, the interaction of proteins in the microenvironment of gastric mucosal atypical hyperplasia was analyzed. The stromata of normal gastric mucosa (NGM) and gastric mucosal atypical hyperplasia (GMAH) tissues were purified with laser capture microdissection (LCM). The differentially expressed GMAH proteins of the NGM and GMAH tissues were identified by quantitative proteomic techniques with isotope labeling. The cross-talk between differentially expressed proteins in NGM and GMAH tissues was then analyzed by bioinformatics. There were 165 differentially expressed proteins identified from the stromata of NGM and GMAH tissues. Among them, 99 proteins were upregulated and 66 were downregulated in GMAH tissue. The present study demonstrated that these proteins in gastric mucosal atypical hyperplasia were involved in cancer-associated signaling pathways, including the p53, mitogen-activated protein kinase (MAPK), cell cycle and apoptosis signaling pathways, and were involved in cellular growth, cellular proliferation, apoptosis and the humoral immune response. The results of the present study suggest that the 165 differentially expressed proteins, including S100 calcium-binding protein A6 (S100A6) and superoxide dismutase 3 (SOD3) in the microenvironment of gastric mucosal atypical hyperplasia, are involved in the p53, MAPK, cell cycle and apoptosis signaling pathways, and serve a function in the pathogenesis of gastric cancer.

Abnormal Expression of PICT-1 and Its Codon 389 Polymorphism Is a Risk Factor for Human Endometrial Cancer

Objective: The protein interacting with carboxyl terminus-1 (PICT-1) gene has been implicated as a tumor suppressor gene, and its alterations have been reported in several cancers. This study investigated the association of PICT-1 alterations with endometrial carcinogenesis. Methods: We analyzed the entire coding region of the PICT-1 gene using polymerase chain reaction-single-strand conformation polymorphism and DNA sequencing to examine PICT-1 mutations in endometrial cancer. Western blotting and immunohistochemical staining were performed to analyze the protein expression and cellular localization of PICT-1 in endometrial cancer cell lines and patient samples. Results: The codon 389 polymorphism of PICT-1 increased the risk of endometrial cancer. Interestingly, 2 of 13 endometrial cancers somatically acquired this mutation compared to normal counterparts. Immunohistochemical staining revealed lower levels of PICT-1 in samples from atypical endometrial hyperplasia and endometrial cancer tissues compared to normal endometrial tissues (p < 0.01). This decrease in PICT-1 expression was significantly correlated with histological grade and lymph node metastasis (p < 0.05). Conclusions: The findings of this study suggest that disruption of PICT-1 protein expression and codon 389 polymorphism can contribute to the pathogenesis or neoplastic progression of endometrial cancer.

Expression and clinical correlation of NGAL and VEGF in endometrial carcinoma.

To determine the expression of neutrophil gelatinase-associated lipocalin (NGAL) and vascular endothelial growth factor (VEGF) in endometrial carcinoma. The clinical features (FIGO staging, pathological type, differentiation level, myometrial invasion depth and tumor size) and immunostaining analysis of endometrial carcinoma (30 cases), atypical hyperplasia (30 cases) and proliferative endometrial tissues (30 cases) were analyzed. Immunohistochemistry and reverse transcription PCR (RT-PCR) were performed to detect the expression of NGAL and VEGF. The positive tissue immunostaining and mRNA expression of NGAL and VEGF in endometrial carcinoma were significantly higher than in either atypical hyperplasia or proliferative endometrial tissues (p<0.05). The relative expression level of NGAL and VEGF was positively correlated with worse FIGO staging, higher differentiation level and a greater myometrial invasion depth (p<0.05); but not with patient age, pathological type or tumor size (p>0.05). The abnormal high expression of NGAL and VEGF observed in the endometrial carcinoma may be an important biomarker for early tumor diagnosis or as a novel target for therapeutic intervention.

Relationship between expression of genes associated with retinoid interferon induced mortality-19, signal transducer and activators of transcription 3, and myeloid cell leukemia-1 with proliferation and apoptosis of esophageal squamous carcinoma cells

Objective To investigate the relationship between the expression of genes associated with retinoid interferon induced mortality-19 (GRIM-19), signal transducer and activators of transcription 3 (STAT3), and myeloid cell leukemia-1 (Mcl-1) with esophageal squamous carcinoma cells proliferation and apoptosis and the possible mechanism. Methods The protein expression of GRIM-19, STAT3, Mcl-1 and proliferation cell nuclear antigen (Ki-67) in 50 cases of esophageal squamous carcinoma tissues, 30 cases of atypical tissue adjacent to carcinoma and 50 cases of normal esophageal mucosa tissues was detected by immunohistochemical streptavidin avidin-peroxidase (SP) method. The mRNA expression of GRIM-19, STAT3, Mcl-1 and Ki-67 in 50 cases of esophageal squamous carcinoma tissues, 30 cases of atypical tissue adjacent to carcinoma and 50 cases of normal esophageal mucosa tissues was detected by in situ hybridization. TdT-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) method was applied to examine apoptosis of esophageal squamous carcinoma cells. Results The positive expression rate of GRIM-19 protein and mRNA in esophageal squamous carcinoma tissues was significantly lower than that in atypical hyperplasia tissue adjacent to carcinoma and normal esophageal mucosa (protein: χ2=12.130, P=0.000; mRNA: χ2=11.830, P=0.000). The positive expression rate of STAT3 and Mcl-1 protein and mRNA in esophageal squamous carcinoma tissues was significantly higher than that in atypical hyperplasia tissue adjacent to carcinoma and normal esophageal mucosa (STAT3 protein: χ2=4.910, P=0.000, STAT3 mRNA: χ2=5.410, P=0.000; Mcl-1 protein: χ2=4.970, P=0.000, Mcl-1 mRNA: χ2=5.370, P=0.000). The apoptosis of tumor cells in esophageal squamous carcinoma tissues was associated with the positive expression rate of GRIM-19, STAT3 and Mcl-1. The apoptosis index (AI) in the group of GRIM-19 positive expression was higher than that in the group of GRIM-19 negative expression (F=18.355, P=0.048). The AI in the group of STAT3 positive expression was lower than that in the group of STAT3 negative expression (F=19.003, P=0.036). The AI in the group of Mcl-1 positive expression was lower than that in the group of Mcl-1 negative expression (F=18.225, P=0.044). Conclusion The down-regulation of GRIM-19, and up-regulation of STAT3 and Mcl-1 may promote esophageal squamous cancer cell proliferation, and inhibit esophageal squamous cancer cell apoptosis. Key words: Genes associated with retinoid interferon induced mortality-19; Signal transducer and activators of transcription 3; Myeloid cell leukemia-1; Esophageal squamous carcinoma cells; Apoptosis

Clinicopathological significance of c-MYC in esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma is one of the most common malignant tumors. The oncogene c-MYC is thought to be important in the initiation, promotion, and therapy resistance of cancer. In this study, we aim to investigate the clinicopathologic roles of c-MYC in esophageal squamous cell carcinoma tissue. This study is aimed at discovering and analyzing c-MYC expression in a series of human esophageal tissues. A total of 95 esophageal squamous cell carcinoma samples were analyzed by the western blotting and immunohistochemistry techniques. Then, correlation of c-MYC expression with clinicopathological features of esophageal squamous cell carcinoma patients was statistically analyzed. In most esophageal squamous cell carcinoma cases, the c-MYC expression was positive in tumor tissues. The positive rate of c-MYC expression in tumor tissues was 61.05%, obviously higher than the adjacent normal tissues (8.42%, 8/92) and atypical hyperplasia tissues (19.75%, 16/95). There was a statistical difference among adjacent normal tissues, atypical hyperplasia tissues, and tumor tissues. Overexpression of the c-MYC was detected in 61.05% (58/95) esophageal squamous cell carcinomas, which was significantly correlated with the degree of differentiation (p = 0.004). The positive rate of c-MYC expression was 40.0% in well-differentiated esophageal tissues, with a significantly statistical difference (p = 0.004). The positive rate of c-MYC was 41.5% in T1 + T2 esophageal tissues and 74.1% in T3 + T4 esophageal tissues, with a significantly statistical difference (p = 0.001). The positive rate of c-MYC was 45.0% in I + II esophageal tissues and 72.2% in III + IV esophageal tissues, with a significantly statistical difference (p = 0.011). The c-MYC expression strongly correlated with clinical staging (p = 0.011), differentiation degree (p = 0.004), lymph node metastasis (p = 0.003), and invasion depth (p = 0.001) of patients with esophageal squamous cell carcinoma. The c-MYC was differentially expressed in a series of human esophageal tissues, and the aberrant c-MYC expression could be a potential factor in carcinogenesis and progression of esophageal squamous cell carcinoma. There was a statistical signification for c-MYC in esophageal squamous cell carcinoma patients to analyze clinicopathological features. It possibly becomes a new diagnostic indicator of esophageal squamous cell carcinoma.

Abstract 4660: HPRT as a biomarker for tumor classification and its variability of expression in breast cancer

Abstract The purpose of this study is to investigate the differential expression of HRPT between various breast cancer tissues in comparison to healthy breast tissue in order to evaluate its potential use as a diagnostic tool. Breast cancer is the most common malignancy diagnosed in women and remains incurable in the metastatic setting requiring accurate and reliable tools for early diagnosis. We analyzed HPRT, a purine salvage pathway enzyme responsible for synthesizing 90% of the available GMP and IMP in cellular maintenance. Due to its critical role in maintaining necessary nucleotides for cell proliferation, we hypothesized an increase in the expression of HPRT in cancer. Tissues from 48 patients with breast carcinoma were stained using standard immunohistochemistry (IHC) techniques. We examined infiltrating ductal carcinoma, margin of carcinoma, hyperplasia, atypical hyperplasia, adenosis, collagen fibers tissues, and normal breast tissue in order to determine expression between various breast cancers. Briefly, tissues were treated with a polyclonal anti-HPRT antibody along with a GADPH positive control and a universal negative control. Tissues were incubated with an HRP-polymer conjugated antibody, and DAP substrate which reacts to antibody-antigen binding sites and results in a color change. Adenosis and hyperplasia are benign breast conditions and are early signs of breast carcinoma development. Hyperplasia tissues, which are in a state of rapid proliferation, showed significant upregulation of HPRT and adenosis tissues showed approximately 42% upregulation. This HPRT upregulation in pre-stage 1 tissues indicates that it may be used as a biomarker to detect early stages of breast cancer. Expression of HPRT was also found to be upregulated in 65% of infiltrating duct carcinoma showing high expression in active malignant areas. In marginal tissue located on the perimeter of primary tumors only 31% of tissues show upregulation of the protein. Because the levels of HPRT in marginal tissue is significantly lower than those found in primary tumors, HPRT may serve as a useful diagnostic tool when evaluating successful tumor removal. These data suggests HPRT as a useful diagnostic tool when identifying tissue with a high proliferation rate indicative of potential cancer development. Additionally, HPRT may also be used as a tool for pathologists to evaluate the success of tumor removal. Citation Format: Michelle H. Townsend, Abigail M. Felsted, Jacob Clarke, Richard A. Robison, Kim L. O'Neill. HPRT as a biomarker for tumor classification and its variability of expression in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4660. doi:10.1158/1538-7445.AM2017-4660

Differences and Relationships Between Normal and Atypical Ductal Hyperplasia, Ductal Carcinoma In Situ, and Invasive Ductal Carcinoma Tissues in the Breast Based on Raman Spectroscopy.

The aim of this study was to find the differences and relationships between normal, atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS), and invasive ductal carcinoma (IDC) lesions of the breast based on biochemical characteristics determined by Raman spectroscopy (RS). After collecting 39 frozen sections from patients who underwent surgical resection or mammotome biopsy, nine normal tissues, seven ADH, eight DCIS, and 15 IDC lesions were detected using confocal RS. We then used leave-one-out cross-validation (LOOCV) and radial basis function (RBF) to build a support vector machine (SVM) diagnosis model. Pronounced mean Raman spectra differences were observed between normal tissues, ADH, DCIS, and IDC tissues. Most noticeable was the increased protein and reduced lipid levels of ADH tissues compared to normal tissues. The major spectra differences in ADH, DCIS, and IDC spectrograms were evidenced by a red shift with a broad peak of CH2 (1301 cm-1), the intensity of the stretching vibration peak of carotenoids (1526 cm-1), a relatively strong band of amide-I (1656 cm-1), and the nuclear (882 cm-1) acid peak. Atypical ductal hyperplasia tissues had the largest constituent variations between subjects. During the disease progression, IDC tissues have smaller inter-subject constituent variations than DCIS and ADH tissues. The overall accuracy of SVM model is 74.39%. The sensitivities of normal tissue, ADH, DCIS, and IDC are 62.5%, 50%, 90%, and 66.7%, respectively. The specificities of normal tissue, ADH, DCIS, and IDC are 100%, 100%, 66.7%, and 89.06%, respectively. Atypical ductal hyperplasia shows significant differences and the relationship between normal tissue and malignant disease. Further study to explain the biochemical relationships between these differences will shed more light into a better understanding of the mechanism by which ADH converts to DCIS and to IDC.

HDAC2 regulates cell proliferation, cell cycle progression and cell apoptosis in esophageal squamous cell carcinoma EC9706 cells.

Increasing evidence has demonstrated that histone deacetylase 2 (HDAC2) participates in the regulation of a variety of biological processes in numerous tumors. However, the potential role of HDAC2 in the development and progression of esophageal squamous cell carcinoma (ESCC) remains elusive. Immunohistochemistry was utilized to detect the expression of HDAC2, Cell Counting Kit-8 was used to determine the cell proliferation, and flow cytometry was employed to investigate cell cycle and cell apoptosis. Finally, western blotting was employed to detect the protein expression of cyclin D1, p21, B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax). The present study found that expression of HDAC2 protein in ESCC tissues was significantly increased compared with atypical hyperplasia tissues and normal esophageal mucosa (P<0.001). The expression of HDAC2 was not associated with the age or gender of patients (P>0.05), but was closely associated with the histological grade, invasion depth, tumor-node-metastasis stage and lymph node metastasis, respectively (all P<0.001). HDAC2 small interfering RNA effectively downregulated the expression of HDAC2 protein in ESCC EC9706 cells. Downregulation of HDAC2 expression evidently inhibited cell proliferation, arrested cell cycle at the G0/G1 phase and induced cell apoptosis in ESCC EC9706 cells, coupled with increased expression of p21 and Bax proteins and decreased expression of cyclin D1 and Bcl-2 proteins. Overall, the present findings suggest that HDAC2 may play an important role in the development and progression of ESCC and be considered as a novel molecular target for the treatment of ESCC.