Relationship between driver gene mutations, their relative protein expressions and survival in non-small cell lung carcinoma in Macao.

OBJECTIVES: We report the status of most common gene mutations in non-small cell lung carcinoma (NSCLC) in Macao, and explore the relationship between each gene mutation and clinicopathologic features and survival. METHODS: EGFR, KRAS and BRAF mutations were detected by PCR in 122 cases of NSCLC. ALK translocation and MET amplification were detected by fluorescence in situ hybridization (FISH). MET and thyroid transcription factor (TTF-1) were investigated by immunohistochemistry. Clinical data were collected for analyzing their correlation with the gene mutations. RESULTS: The mutation of EGFR, KRAS and BRAF was detected in 48 (39.3%), 13 (10.7%) and 3 (2.5%) of 122 cases of NSCLC, respectively. ALK translocation and MET amplification were detected in 7 (5.7%) and 3 cases (2.5%). The rate of EGFR mutation was significantly higher in female and non-smoker patients. In TTF-1 positive cases EGFR mutation was more frequent. Age of the patients over 62-year old was correlated with KRAS mutations. The concordance between ALK IHC and FISH was 58.3%. The MET protein in the cases with MET amplification was 100% positive. The survival was lower in the patients with positive MET protein than those with negative. MET protein was an independent prognostic factor for NSCLC. CONCLUSIONS: EGFR mutation occurred frequently in the female never smoke patients with NSCLC. KRAS mutation was more common in old patients. Negative MET protein expression could be used as a negative predictive marker of MET amplification. MET protein expression was an independent prognostic factor for NSCLC.

Collision of lymphoepithelioma-like carcinoma and adenocarcinoma of the lung: a case report.

We report a rare case of collision of lymphoepithelioma-like carcinoma (LELC) and adenocarcinoma (AC) in the lung. A 59-year-old woman had a history of fever and cough. A mass was found by X-ray in the left upper lung. Magnetic resonance imaging (MRI) shows a dumbbell-like mass in the fore and tongue segment of the left upper lung with irregular spiculate margin. Positron emission tomography/computed tomography (PET/CT) (18F-FDG) shows strong concentration of radioactivity (SUVmax 6.9-12.3 cm) in the lung mass only. The patient subsequently underwent resection of left upper lung and associated hilar lymph nodes. Histological examination revealed it was a collision carcinoma comprising LELC and AC. The hilar lymph nodes were tumuor free. The immunoreactions, Epstein-Barr early RNA in situ hybridization and molecular analyses, such as EGFR mutation, c-Met, anaplastic lymphoma kinase were different in both tumuor components, indicating they derived from different cell origin. This rare case was discussed.

Protein expression of HER2, 3, 4 in gastric cancer: correlation with clinical features and survival.

BACKGROUND AND AIMS: Despite significant improvements in targeted therapies for patients with advanced gastric cancer (GC), the prognosis of those patients remains poor. This study explores the expression and clinicopathological significance of human epidermal growth factor receptor 2, 3 and 4 (HER2, HER3, HER4) in GC, in order to find more prognostic biomarkers of GC and putative targets of therapy. METHODS: Immunohistochemistry was performed for HER2, HER3 and HER4 in 498 patients with GC using tissue microarray. Correlations between the receptor expression and clinicopathological features, as well as prognosis of the patients were statistically analysed. RESULTS: The high expression rates of HER2, HER3 and HER4 proteins in the patients were 8.6% (43/498), 20.7% (103/498) and 13.3% (66/498), respectively. High expression of HER2 and HER3 was correlated with proximal GC of the cardia (p<0.05). High expression of HER3 was associated with the tumour depth, tumour node metastasis (TNM) stage and lymph node metastasis (p<0.05). High expression of HER4 was associated with TNM stage (p<0.05) only. According to a regression model, high expression of HER3 was significantly associated with patients' poor survival (p=0.004). High expression rates of HER2, HER3 and HER4 were correlated with each other, but they were all associated merely with histologically intestinal-type adenocarcinoma of GC (p<0.05). CONCLUSIONS: HER3 is correlated with the malignant biological behaviour of GC. Expression of HER3 is a significant predictor of poor survival in GC. Therefore, the development of HER3-targeted agents may provide new possibilities in the treatment of GC.

Hepatocellular carcinoma with bile duct tumor thrombus: a clinicopathological analysis of factors predictive of recurrence and outcome after surgery.

Although hepatocellular carcinoma (HCC) with bile duct tumor thrombus (BDTT) is a rare entity, most patients experience tumor recurrence even after curative resection and the prognosis remains dismal. This study aimed to analyze the clinicopathological risk factors for recurrence and poor outcome after surgical treatment of HCC with BDTT.Clinicopathological data of 37 patients with HCC and BDTT who underwent surgical treatment from July 2005 to June 2012 at the authors' hospital were reviewed retrospectively. Prognostic factors and potential risk factors for recurrence were assessed by Cox proportional hazard model and binary logistic regression model, respectively.Among the 37 patients, anatomical and nonanatomical liver resection was performed in 26 and 11 patients, respectively. The resection was considered curative in 19 patients and palliative in 18 patients. Also, 21 cases had tumor recurrence after operation and 7 cases of them were reoperated. Multivariate binary logistic regression model revealed that surgical curability was the only independent risk factor associated with postoperative tumor recurrence (P = 0.034). In addition, postoperative overall survival rates at 1, 2, and 3 years were 64.2%, 38.9%, and 24.3%, respectively. Cox multivariate analysis indicated that surgical curability and tumor recurrence were independent prognostic factors for both overall survival and recurrence-free survival (P < 0.05).Although patients with HCC and BDTT had a relatively high rate of early recurrence after surgery, relatively favorable long-term outcome after curative hepatic resection could be achieved. Therefore, extensive and curative surgical treatment should be recommended when complete resection can be achieved and liver functional reserve is satisfactory.

[Use of extend lateral upper arm free flap for reconstruction of soft tissue defect after resection of oral cancer].

PURPOSE: To investigate the clinical outcomes of composite lateral arm free flap to reconstruct soft tissue defect after oral cancer ablation. METHODS: Fifteen cases of soft-tissue defects were reconstructed using composite lateral arm free flap from January 2011 to December 2013. Color Doppler ultrasound blood flow detector was used to avoid variation of posterior radial collateral artery (PRCA). Composite flaps were designed according to the direction of PRCA, shape and area of the soft-tissue defect, which were over condylus lateralis humeri, with areas ranged from 4 cｍ×5 cm-11 cm×5 cm and an average vascular pedicle length of 10cm. Primary closure was achieved on all donor sites. RESULTS: Fourteen flaps survived while vascular crisis was found in one flap 1 day post operation. The flap was replaced after an invalid salvation. All patients were followed up over 12 months, no recurrence was found. All patients achieved good recovery of morphology and functions with only 2 reporting local numbness of donor sites, which were relieved after 6 months. CONCLUSIONS: Composite lateral arm free flap, with consistent anatomy, suitable thickness and reliable blood supply, is a proper method to reconstruct postoperative soft tissue defect of patients with oral carcinoma.

Expression of CD44v3, erythropoietin and VEGF-C in gastric adenocarcinomas: correlations with clinicopathological features.

AIMS: To investigate the expression of CD44v3 and vascular endothelial growth factor C (VEGF-C) in gastric adenocarcinoma and the correlation with erythropoietin (EPO) and clinicopathological features. METHODS: The expression of CD44v3, VEGF-C and EPO was evaluated by immunohistochemical staining in 169 gastric adenocarcinomas. The correlations between these parameters and patients' clinicopathological features were analyzed statistically. RESULTS: CD44v3 and VEGF-C were positive in 50 (29.6%) and 82 (48.5%) patients, respectively. High CD44v3 expression was associated with poor cellular differentiation, extensive lymph node metastasis and advanced stage of gastric adenocarcinoma (P <0.05). High VEGF-C expression was significantly correlated with Lauren type, extensive lymph node metastasis and advanced stage of gastric adenocarcinoma (P <0.05). Univariate analysis of survival demonstrated that patients with a strong CD44v3 immunoreaction had a significantly worse overall survival compared with patients showing a weak CD44v3 immunoreaction (log-rank test: P = 0.0449). The mean survival time of patients with low CD44v3 expression was 30.6 months, which was much longer than the 21.6 months in patients with high expression. In addition, a strong association between immunohistochemical expression of CD44v3 and EPO was noted. CONCLUSION: Increased expression of CD44v3 and VEGF-C may play a significant role in the carcinogenesis and progression of gastric adenocarcinoma. High CD44v3 expression may be a predictor of poor prognosis in gastric adenocarcinoma patients. We infer that some mechanisms may exist in regulating the expression of CD44v3 and EPO.

Proteins related to early changes in carcinogenesis of hepatic oval cells after treatment with methylnitronitrosoguanidine.

Hepatic oval cells are considered as facultative progenitor/stem cells of liver and able to differentiate into either hepatocytes or biliary epithelial cells. The transformed oval cells by carcinogen possess potential to develop carcinomas in animal models. In order to better understand the molecular mechanism in carcinogenetic process, we used a proteomic approach to assess the early changes in protein expression of oval cells (OC3W3-15) initiated by methylnitronitrosoguanidine (MNNG). Meanwhile, we compared cell biologic characteristics of the MNNG treated OC3W3-15 cells and control oval cells by electron microscopy, flow cytometry, karyotype and soft agar assay. The mRNA levels of GGT and GSTP1 determined by real-time PCR were also detected in both cell lines. Our results showed that MNNG-treated OC3W3-15 cells exhibited characteristics of malignant transformation, including growth rate, chromosomal aberrations, abnormal DNA content, and the ability to form colonies. The cells expressed higher levels of the tumor marker AFP, GGT and GSTP1 mRNA than that of control cells. Significant changes of several proteins involved in the malignant transformation process, including cell cycle related proteins, proteins involved in organism development and cell differentiation, are found in OC3W3-15 cells. The proteins may provide early affection in malignant transformation of hepatic oval cells, and yield further insight into mechanism of carcinogenesis of hepatocellular carcinoma.

Regulation effect of zinc fingers and homeoboxes 2 on alpha-fetoprotein in human hepatocellular carcinoma.

Aim. To investigate the relationship between alpha-fetoprotein and zinc fingers and homeoboxes 2 in hepatocellular carcinoma. Materials and Methods. The expressions of zinc fingers and homeoboxes 2, nuclear factor-YA, and alpha-fetoprotein mRNA in 63 hepatocellular carcinoma were detected by reverse transcriptase-polymerase chain reaction and compared with the clinical parameters of the patients. Selectively, silence of zinc fingers and homeoboxes 2 in HepG2 cells was detected by RNA interference technique. Results. Alpha-fetoprotein mRNA expression was detected in 60.3% of hepatocellular carcinoma cases. Zinc fingers and homeoboxes 2 mRNA expression (36.5%) was significantly negatively correlated with serum alpha-fetoprotein concentration and mRNA expression. A strong positive correlation was found between zinc fingers and homeoboxes 2 and nuclear factor-YA mRNA expression (42.9%), while the latter was negatively correlated with serum alpha-fetoprotein concentration and mRNA expression. Treatment with zinc fingers and homeoboxes 2 small interfering RNA led to 85% and 83% silence of zinc fingers and homeoboxes 2 mRNA and protein expression and 60% and 61% reduction of nuclear factor-YA mRNA and protein levels in the HepG2 cells, respectively. Downregulation of zinc fingers and homeoboxes 2 also induced a 2.4-fold increase in both alpha-fetoprotein mRNA and protein levels. Conclusions. Zinc fingers and homeoboxes 2 can regulate alpha-fetoprotein expression via the interaction with nuclear factor-YA in human hepatocellular carcinoma and may be used as an adjuvant diagnostic marker for alpha-fetoprotein-negative hepatocellular carcinoma.

[Glypican 3 expression in hepatoblastoma and its diagnostic implication].

OBJECTIVE: To explore the expression and diagnostic significance of glypican-3 (GPC3) in hepatoblastoma. METHODS: Five tissue microarray paraffin blocks were constructed to include 54 cases of hepatoblastoma. The tumor tissue samples were obtained from 3 surgical biopsies, 33 needle biopsies, 5 stage I resection tumors, and 13 stage II resection tumors after transcatheter arterial chemoembolization. Ten samples of non-neoplastic hepatic tissue adjacent to tumor were used as control. Immunohistochemical staining of GPC3 (clone 1G12) was performed. Among the 54 cases of hepatoblastoma, 22 cases were fetal subtype, 24 cases were mixed fetal and embryonal subtype and 8 cases were mixed epithelial and mesenchymal type. RESULTS: GPC3 was positive in fetal epithelial cells (54/54, 100%), but negative or weakly positive in embryonic epithelial cells in all cases of hepatoblastoma. Undifferentiated small cells and all mesenchymal components were negative for the expression. Non-neoplastic hepatocytes adjacent to tumor were negative for GPC3 expression (0/10) . CONCLUSIONS: Fetal epithelial components of hepatoblastoma express GPC3 protein detectable by immunohistochemistry. Normal hepatocytes after birth, small cell undifferentiated and embryonic epithelial components of hepatoblastoma do not or weakly express GPC3 protein. Therefore, GPC3 immunohistochemistry offers a valuable aid to the diagnosis of hepatoblastoma in infants and children.

Recurrent meningeal hemangiopericytoma with multiple metastasis and hypoglycemia: a case report.

Aims. We report on the unusual case of a 43-year-old man who developed recurrent meningeal hemangiopericytoma and presented with hypoglycemia 6 years after excision of the tumor. Methods and Results. We utilized computed tomography to assure multiple tumor metastasis and cranial recurrence of previous meningeal hemangiopericytoma and clinical laboratory tests and immunohistochemical staining to characterize this case. Magnetic resonance imaging and computed tomography showed the recurrent tumor at original torcular site was increased in size. Abnormal low levels of growth hormone, insulin, and insulin-like growth factor-I except insulin-like growth factor-II were detected in the serum. By immunohistochemistry, the neoplastic cells characteristically express diffusely CD99, bcl2, and variable CD34. After radio- and chemotherapy, serum glucose level of the patient returned to normal. Conclusions. Comparing other brain tumors, meningeal hemangiopericytoma has a higher recurrent and metastatic rate, but this tumor with hypoglycemia is very rare.

[Amplification of HER2 gene in gastric carcinoma detected by dual in-situ hybridization].

OBJECTIVE: To explore the status of HER2 gene amplification and its product HER2 protein expression in gastric carcinoma, so as to aid in patient selection for anti-HER2 targeted chemotherapy. METHODS: Eighty-five cases of gastric carcinoma biopsy tissues were collected. The status of HER2 gene amplification was detected by dual in situ hybridization (dual-ISH). And HER2 protein was detected by immunohistochemistry. RESULTS: HER2 gene amplification was detected in 10/85 (11.8%) cases of gastric carcinoma, and no amplification was detected in 75/85 (88.2%) cases. In the 10 cases with HER2 amplification, HER2 immunoreaction scorings of 3+, 2+ and 0/1+ were present in 7, 2 and 1 cases, respectively. In the 75 cases without HER2 amplification, HER2 immunoreaction scorings of 3+, 2+ and 0/1+ were present in 0, 18 (24.0%) and 57 (76.0%) cases, respectively. Histologically, most gastric carcinoma with amplification of HER2 gene was moderately differentiated tubular adenocarcinoma. CONCLUSIONS: HER2 gene dual-ISH technique is a reliable and objective method for detecting HER2 gene amplification in gastric carcinoma biopsy. Clinically, only few gastric carcinomas show HER2 gene amplification and are suitable candidates for anti-HER2 targeted chemotherapy.

[Significance of glypican-3 immunohistochemistry in diagnosis of hepatocellular carcinoma].

OBJECTIVE: To explore the diagnostic significance of glypican-3 (GPC3) immunohistochemistry in hepatocellular carcinoma (HCC). METHODS: Fourteen tissue microarray paraffin blocks were constructed, which comprised 731 samples from hepatic tumors and paratumor tissues, including 357 cases of HCC, 26 cholangiocarcinoma, 171 HCC adjacent hepatic tissue including cirrhosis, 93 hemangioma adjacent hepatic tissues, and 84 carcinomas metastatic to liver. GPC3 (Clone 1G12) protein was detected immunohistochemically in all of cases with positive controls. RESULTS: GPC3 protein was positive in 72.0% HCC (257/357), but negative in the rest 374 of non-HCC cases, including cholangiocarcinoma, HCC adjacent hepatic tissue including cirrhosis, hemangioma adjacent hepatic tissues and metastatic carcinomas. GPC3 positive percentage was significantly correlated with histological grading of HCC (P < 0.01), highest in grade 3 (77.1%, 64/83) followed by grade 2 (73.3%, 187/255), grade 1 (6/12) and grade 4 (0). CONCLUSIONS: GPC3 is a valuable diagnostic marker for hepatocellular carcinoma with sensitivity of 72.0%, and a differential diagnostic marker from tumor adjacent hepatic tissue and carcinomas metastatic to liver with specificity of 100%.

Prognostic significance of erythropoietin and erythropoietin receptor in gastric adenocarcinoma.

AIM: To investigate the expression of Erythropoietin (Epo) and its receptor (EpoR) in gastric adenocarcinoma (GAC) and the correlation with angiogenesis and clinicopathological features. METHODS: The expressions of Epo, EpoR and vascular endothelial growth factor (VEGF), as well as microvessel density were evaluated in 172 GAC biopsies by immunohistochemical staining. The correlations between these parameters and patient's clinicopathological features were analyzed statistically. RESULTS: The proportion of Epo and EpoR alterations in GAC was higher than that in adjacent normal mucosa (P = 0.035 and 0.030). Epo high-expression was associated with EpoR high-expression, Lauren type, extensive lymph node metastasis and advanced stage of GAC (P = 0.018, 0.018, 0.004 and 0), while EpoR expression was linked with older age, World Health Organization type, extensive lymph node metastasis and advanced stage (P = 0.001, 0.013, 0.008 and 0.001). VEGF high expression was significantly correlated with EpoR low-expression, Lauren type, extensive lymph node metastasis and advanced stage (P = 0.001, 0.001, 0.001 and 0.007). The expression of Epo or EpoR was associated with microvessel density (P = 0.004 and 0.046). On multivariate analysis, only lymph node metastasis, abnormal Epo expression and tumor nodes metastases stage were independently associated with survival. In addition, a strong association with the immunohistochemical expression of EpoR and the angiogenic protein, VEGF, was noted. CONCLUSION: Increased expression of Epo and EpoR may play a significant role in the carcinogenesis, angiogenesis and progression of GAC. Epo may be an independent prognostic factor.

Diagnostic utility of glypican-3 for hepatocellular carcinoma on liver needle biopsy.

AIMS: Glypican 3 (GPC3) is a heparan sulphate proteoglycan that shows elevated levels in the serum of patients with hepatocellular carcinoma (HCC), but not in healthy blood donors or patients with benign liver disease. This study explores the value of GPC3 expression for diagnosis of HCC by immunohistochemistry in liver needle biopsy specimens. METHODS: Archival material of liver needle biopsies from 54 patients with HCC, nine with focal nodular hyperplasia or focal liver cell dysplasia, five with cirrhosis, seven with hepatitis B or unremarkable liver tissue, seven with cholangiocarcinoma, and 30 with metastatic tumours, was retrieved for immunohistochemical staining with GPC3 antibody and appropriate positive and negative controls. RESULTS: Forty-five out of 54 cases of HCC showed positive GPC3 staining (83.4%). In contrast, all 58 non-HCC cases of liver biopsies, including focal nodular hyperplasia, focal liver cell dysplasia, cirrhosis, hepatitis B or unremarkable liver tissue, cholangiocarcinoma and metastatic tumours, were negative for GPC3. The sensitivity and specificity of GPC3 in HCCs were 83.4% and 100%, respectively. CONCLUSIONS: GPC3 is a valuable diagnostic marker for diagnosing HCC on liver needle biopsy. It can be used to distinguish HCC from other benign hepatic conditions and metastatic tumours in the liver.

Overexpression of YKL-40 is an independent prognostic marker in gastric cancer.

YKL-40 is a growth factor for connective tissue cells and a migration factor for endothelial cells. Elevated serum level of YKL-40 has been associated with poor prognosis in many cancers. However, the status of YKL-40 expression and its clinical/prognostic significance in gastric cancer are unclear. In this study, the expression of YKL-40 was studied by immunohistochemistry in gastric cancer tissue microarray containing 172 primary gastric cancer cases and 70 adjacent nonneoplastic mucosa specimens. The correlations between YKL-40 expression and clinicopathologic features, as well as activation of PI3K/Akt pathways were addressed. Expression of YKL-40 was significantly higher in gastric cancer tissues than that in adjacent nonneoplastic tissues. Overexpression YKL-40 was found in 28.4% of gastric cancers and was significantly associated with tumor invasion (P = .007) and lymph node metastasis (P = .009). For survival study, overexpression of YKL-40 was significantly associated with worse outcome (P = .001). When known clinical variables were added to a multivariate analysis, TNM stage, tumor size, and overexpression of YKL-40 emerged as independent prognostic factors. Further study indicated that the oncogenic function of YKL-40 might be through the activation of Akt pathway. These results suggest that overexpression of YKL-40 is correlated with the aggressive behavior of tumor cells, which could be used as an independent molecular marker for the predicting poor prognosis of patients with gastric cancer.

Downregulation of ZIP kinase is associated with tumor invasion, metastasis and poor prognosis in gastric cancer.

Deletion of 19p13 is one of the most frequent genetic changes in gastric carcinoma (GC), implying the existence of a tumor suppressor gene (TSG) that plays an important role in GC development. To identify the candidate TSG at 19p, array-comparative genomic hybridization (CGH) was applied to study DNA copy-number changes on chromosomes 3, 5p, 13, 16q and 19. The result showed that gains of 16q21, 19q13.1, 5p15.1 and 3q26.31, and losses of 3p21.32, 3p22.2, 19q13.33 and 19p13.3, were frequently detected by array-CGH. One candidate TSG, ZIP kinase (ZIPK), at 19p13.3 was further characterized by immunohistochemistry using a tissue microarray containing 172 primary GCs. Downregulation of ZIPK was detected in 111/162 informative GCs, which was significantly associated with invasion, metastasis and poorer prognosis of GC. To investigate the association of the downregulation of ZIPK with apoptosis, apoptosis assay (TUNEL) was used to compare the apoptotic index between GCs with normal expression and downregulation of ZIPK. TUNEL assay showed that the apoptotic index in GCs with normal ZIPK expression was significantly higher than that in GCs with downregulation of ZIPK (p < 0.001), indicating that ZIPK plays an important pro-apoptotic role in GC. Taken together, we demonstrated here that ZIPK is a tumor suppresser gene and plays an important role in GC development through its pro-apoptotic function. Downregulation of ZIPK can be used to evaluate tumor invasiveness, metastasis and to predict survival of GC.

An intrasellar mixed gangliocytoma-adenoma including ependymal component, and review of the literature.

A rare case of intrasellar mixed gangliocytoma-adenoma including ependymal component, which presented clinically with acromegaly and menstrual disorder, is described here. The tumour was totally removed via trans-sphenoidal surgery. A histological examination of the resected specimen showed that the tumour was composed of ganglion cells, adenomatous cells and ependymal cells. Most intrasellar gangliocytomas are composed of two components: adenomatous cells and ganglion cells. In this case, in addition to mixed adenomatous and ganglion cells, focal ependymal cells forming small cysts were found. Based on these histopathological findings, it was inferred that stem cells existed in the pituitary gland during embryonic development. The stem cells were able to differentiate into three directions: ganglion cells, adenomatous cells and ependymal cells, and as a result an intrasellar mixed gangliocyto-adenoma including ependymal component developed.

[Role of AIB1 gene in development of esophageal squamous cell carcinoma and its clinicopathologic significance].

OBJECTIVE: To investigate the role of AIB1 gene in the development of esophageal squamous cell carcinoma (ESCC) and its clinicopathologic significance. METHODS: Two tissue microarrays, including 203 cases of ESCC, were prepared. Fluorescence in-situ hybridization (FISH) and immunohistochemistry were performed to detect the amplification of AIB1 gene and expression of its encoded protein in ESCC. The results were correlated with various clinicopathologic parameters. RESULTS: In the 203 cases of ESCC studied, FISH was successful in 115 cases. Amongst those, amplification/gain of AIB1 gene was observed in 15 cases, including high-level amplification in 5 cases (4.3%) and low-level gain in 10 cases (8.7%). As for immunohistochemical study, AIB1 protein was overexpressed in 94 cases of ESCC. There was a significant association of AIB1 overexpression and tumor staging. AIB1 was overexpressed in 66 of the 123 cases in advanced T stages (T3 to 4), compared with 25 of the 80 cases in early T stages (P = 0.008). Those cases with high-level amplification of AIB1 also showed overexpression of its encoded protein. On the other hand, 8 of the 10 cases with low-level gain of AIB1 showed protein overexpression. The remaining 41 of the 100 cases which did not have AIB1 gene amplification/gain demonstrated overexpression of AIB1 protein. CONCLUSION: Overexpression of AIB1 protein caused by gene amplification/gain or other molecular mechanisms may play an important role in the development and/or progression of a subset of ESCC.