Jing Hui Hou

miR-125b is Methylated and Functions as A Tumor Suppressor by Regulating the ETS1 proto-oncogene in Human Invasive Breast Cancer

The microRNA miR-125b is dysregulated in various human cancers but its underlying mechanisms of action are poorly understood. Here, we report that miR-125b is downregulated in invasive breast cancers where it predicts poor patient survival. Hypermethylation of the miR-125b promoter partially accounted for reduction of miR-125b expression in human breast cancer. Ectopic restoration of miR-125b expression in breast cancer cells suppressed proliferation, induced G(1) cell-cycle arrest in vitro, and inhibited tumorigenesis in vivo. We identified the ETS1 gene as a novel direct target of miR-125b. siRNA-mediated ETS1 knockdown phenocopied the effect of miR-125b in breast cell lines and ETS1 overexpression in invasive breast cancer tissues also correlated with poor patient prognosis. Taken together, our findings point to an important role for miR-125b in the molecular etiology of invasive breast cancer, and they suggest miR-125b as a potential theranostic tool in this disease.

Elevated expressions of survivin and VEGF protein are strong independent predictors of survival in advanced nasopharyngeal carcinoma

BackgroundNasopharyngeal carcinoma (NPC) is one of the most common malignancies in southern China. The China 1992 TNM staging system has been widely used for prognosis prediction of NPC patients in China. Although NPC patients can be classified according to their clinical stage in this system, their prognosis may vary significantly.Method280 cases of NPC with clinical follow-up data were collected and expressions of survivin and VEGF in tumor tissues were investigated by immunohistochemistry (IHC). Apoptosis index (AI) in 100 cases of NPC was detected by the TUNEL method.ResultsExpression of survivin and VEGF were significantly associated with TNM stage, T-stage and metastasis of NPC. The patients with survivin and VEGF over-expression presented lower 5-year survival rate, as compared to those of low-expression (42.32% vs. 70.54%, 40.1% vs. 67.8%, respectively, P < 0.05), especially in advanced stage patients (36.51% vs. 73.41%, 35.03% vs. 65.22%, respectively, P < 0.05). The 5-year survival rate in NPC patients with survivin and VEGF dual over-expression was significantly lower than that of patients with dual low-expression (18.22% vs. 73.54%, respectively; P = 0.0003). Multivariate analysis indicated that both survivin and VEGF over-expression in NPC tumor tissues were strong independent factors of poor prognosis in NPC patients. The mean AI in the 39 survivin low-expression cases was 144.7 ± 39.9, which was significantly higher than that in 61 survivin over-expression cases (111.6 ± 39.8) (T test, P < 0.05).ConclusionSurvivin and VEGF over-expression are independent prognostic factors for the patients with NPC. These results also suggest that tumor survivin and VEGF expressions are valuable prognostic markers for prognosis prediction in NPC patients.

The density of macrophages in the invasive front is inversely correlated to liver metastasis in colon cancer

BackgroundAlthough an abundance of evidence has indicated that tumor-associated macrophages (TAMs) are associated with a favorable prognosis in patients with colon cancer, it is still unknown how TAMs exert a protective effect. This study examined whether TAMs are involved in hepatic metastasis of colon cancer.Materials and methodsOne hundred and sixty cases of pathologically-confirmed specimens were obtained from colon carcinoma patients with TNM stage IIIB and IV between January 1997 and July 2004 at the Cancer Center of Sun Yat-Sen University. The density of macrophages in the invasive front (CD68TFHotspot) was scored with an immunohistochemical assay. The relationship between the CD68TFHotspot and the clinicopathologic parameters, the potential of hepatic metastasis, and the 5-year survival rate were analyzed.ResultsTAMs were associated with the incidence of hepatic metastasis and the 5-year survival rate in patients with colon cancers. Both univariate and multivariate analyses revealed that the CD68TFHotspot was independently prognostic of survival. A higher 5-year survival rate among patients with stage IIIB after radical resection occurred in patients with a higher macrophage infiltration in the invasive front (81.0%) than in those with a lower macrophage infiltration (48.6%). Most importantly, the CD68TFHotspot was associated with both the potential of hepatic metastasis and the interval between colon resection and the occurrence of hepatic metastasis.ConclusionThis study showed evidence that TAMs infiltrated in the invasive front are associated with improvement in both hepatic metastasis and overall survival in colon cancer, implying that TAMs have protective potential in colon cancers and might serve as a novel therapeutic target.

High expression of H3K27me3 in human hepatocellular carcinomas correlates closely with vascular invasion and predicts worse prognosis in patients

It has been suggested that trimethylation of lysine 27 on histone H3 (H3K27me3) is a crucial epigenetic process in tumorigenesis. However, the expression dynamics of H3K27me3 and its clinicopathological/prognostic significance in hepatocellular carcinoma (HCC) are unclear. In this study, immunohistochemical analysis (IHC) was used to examine protein expression of H3K27me3 in HCC tissues from two independent cohorts and corresponding nontumorous hepatocellular tissues by tissue microarray. The optimal cutpoint of H3K27me3 expression was assessed by the X-tile program. Our results showed that the cutpoint for high expression of H3K27me3 in HCCs was determined when more than 70% of the tumor cells showed positive staining. High expression of H3K27me3 was observed in 134 of 212 (63.2%) and 76 of 126 (60.4%) of HCCs in the testing and validation cohorts, respectively. Correlation analysis demonstrated that high expression of H3K27me3 in HCCs was significantly correlated with large tumor size, multiplicity, poor differentiation, advanced clinical stage and vascular invasion (P < 0.05). In addition, high expression of H3K27me3 in HCC patients was associated closely with shortened survival time, independent of serum α-fetoprotein levels, tumor size and multiplicity, clinical stage, vascular invasion and relapse as evidenced by univariate and multivariate analysis in both cohorts (P < 0.05). In different subsets of HCC patients, H3K27me3 expression was also a prognostic indicator in patients with stage II tumors (P < 0.05). Thus, these findings provide evidence that a high expression of H3K27me3, as detected by IHC, correlates closely with vascular invasion of HCCs and is an independent molecular marker for poor prognosis in patients with HCC.

Overexpression of MMP-2 in laryngeal squamous cell carcinoma: a potential indicator for poor prognosis.

OBJECTIVES: To investigate the expression and clinical significance of gelatinases (MMP-2 and MMP-9) in patients with laryngeal squamous cell carcinoma (LSCC). STUDY DESIGN AND SETTING: In a retrospective study of 72 consecutive patients with LSCC hospitalized in a single cancer center, immunohisto-chemistry was used to examine the expression of MMP-2 and MMP-9 in surgical samples. The results were compared to clinicopathological features and prognosis. RESULTS: The positive expression of MMP-2 and MMP-9 in patients with LSCC was 50% (36/72) and 73.6% (53/72), respectively. According to the expression scale, there were 36 patients of -, 26 patients of +, 7 patients of + +, and 3 patients of + + + expression of MMP-2; 19 patients of -, 26 patients of +,16 patients of ++, and 11 patients of + + + expression of MMP-9. There was no significant relationship found between the expression of MMP-2 or MMP-9 and clinicopathological features of LSCC, such as histological grade, primary site, T stage, N stage, and clinical stage. The 5-year overall survival (OS) and disease-free survival (DFS) rate calculated by Kaplan-Meier method in patients with negative and positive expression of MMP-9 and MMP-2 was 73.68%, 50.94%, 73.68%, and 49.06% in MMP-9 and 72.22%, 41.67%, 72.22%, and 38.89% in MMP-2, respectively. Significant 5-year survival difference was found between patients with negative and positive expression of MMP-2 (log rank = 6.74, P = 0.0094). There was significant lower survival rate in patients with higher positive expression of MMP-2 (log rank = 11.77, P = 0.0028). In glottic laryngeal cancer, positive expression of MMP-2 could predict poor survival and was more likely to present primary recurrence. CONCLUSION: The expression of MMP-2 could be used as a potential predictor for poor prognosis in patients with LSCC.

Epstein-Barr virus encoded latent membrane protein 1 regulates mTOR signaling pathway genes which predict poor prognosis of nasopharyngeal carcinoma

BackgroundThe oncoprotein Epstain-Barr Virus (EBV)-encoded latent membrane protein1 (LMP1) modulates the pathological effects of the NF-κB, AP-1 and JAK/STAT pathways in nasopharyngeal carcinoma (NPC).MethodsMicroarray analysis was performed on the NPC cell line HONE1 stably transfected with a LMP1-expression plasmid or an empty vector. Based on assigned pathways analyzed using the KEGG database, the mTOR signaling pathway was selected for verification by quantitative RT-PCR. Western blot, RNA interference and immunofluorescence were used to determine the relationship between LMP1 and mTOR signing pathway genes, and their clinical significance to NPC.ResultsOur studies revealed that overexpression of LMP1 upregulated the mTOR signaling pathway, possibly through phosphorylation of AKT/mTOR/P70S6K/4EBP1 in the NPC cell lines HONE1 and 6-10B. Knockdown of LMP1 reduced expression of p-mTOR and p-4EBP1 in EBV-positive NPC cell line C666-1. In addition, LMP1 expression closely correlated with expression of p-mTOR, p-P70S6K and p-4EBP1 in NPC tumors. Expression of p-P70S6K, p-4EBP1 and LMP1, but not p-mTOR, significantly correlated with overall survival of NPC patients. However, only LMP1 was an independent prognostic factor.ConclusionsThese results suggest that the mTOR signaling pathway is regulated by LMP1 expression in NPC. LMP1 and the genes in the mTOR pathway such as p-P70S6K and p-4EBP1 may be potential prognostic biomarkers.

Secreted protein acidic and rich in cysteine (SPARC) is associated with nasopharyngeal carcinoma metastasis and poor prognosis

BackgroundThe aim of the present study was to analyse the expression of Secreted protein acidic and rich in cysteine (SPARC) in nasopharyngeal carcinoma (NPC) specimens, and to evaluate its correlation with clinicopathologic features, including survival of patients with NPCMethodsNPC tissue microarrays (TMAs) were constructed from Sun Yat-sen University Cancer Center (SYSUCC), another three centers on mainland China, Singapore and Hong Kong. Using quantitative RT-PCR and Western-blotting techniques, we detected mRNA and protein expression of SPARC in NPC cell lines and immortalized nasopharyngeal epithelial cells (NPECs) induced by Bmi-1 (NPEC2 Bmi-1). The difference of SPARC expression in the cell lines was tested using a t-test method. The relationship between the SPARC expression and clinicopathological data was assessed by chi-square. Survival analysis was estimated using the Kaplan-Meier approach with log-rank test. Univariate and multivariate analyses of clinical variables were performed using Cox proportional hazards regression models.ResultsThe expression levels of SPARC mRNA and protein were markedly higher in NPC cell lines than in NPEC2 Bmi-1. Especially, the expression levels of SPARC mRNA and protein were much lower in the 6-10B than in the 5-8 F (P = 0.002, P = 0.001). SPARC immunostaining revealed cytoplasmic localization in NPC cells and no staining in the stroma and epithelium.In addition, high level of SPARC positively correlated with the status of distant metastasis (P = 0.001) and WHO histological classification (P = 0.023). NPC patients with high SPARC expression also had a significantly poorer prognosis than patients with low SPARC expression (log-rank test, P < 0.001), especially patients with advanced stage disease (log-rank, P < 0.001). Multivariate analysis suggested that the level of SPARC expression was an independent prognostic indicator for the overall survival of patients with NPC (P < 0.001).ConclusionsSPARC expression is common in NPC patients. Our data shows that elevated SPARC expression is a potential unfavorable prognostic factor for patients with NPC.

nm23-H1 expression in nasopharyngeal carcinoma: Correlation with clinical outcome

The expression levels of nm23‐H1 mRNA and its protein in human nasopharyngeal carcinoma (NPC) were detected to clarify the relationship between nm23‐H1 and metastasis and prognosis of patients with NPC. nm23‐H1 mRNA expression in fresh tissues from 78 patients with NPC was investigated by in situ hybridization and RT‐PCR. Routine labeling streptavidin‐biotin immuno‐histochemistry with the nm23‐H1 murine monoclonal antibody was employed to study the expression of nm23‐H1 protein in paraffin‐embedded specimens from 231 patients with NPC treated in our hospital. The clinical pathologic data and results of follow‐up were collected. Comparisons between expression of nm23‐H1 protein or mRNA and clinical outcome were performed using the χ2 test. Multivariate prognostic analyses were performed by the Cox regression model. We found that nm23‐H1‐negative tumors were associated with a higher incidence of lymph‐node metastasis (84.2%) than nm23‐H1‐positive ones (32.8%, p < 0.01). The distant metastasis and loco‐regional recurrence rates in the nm23‐H1‐negative group were 55.8% and 31.68%, respectively but only 17.2% and 11.5%, respectively, in the nm23‐H1‐positive group (p < 0.01). A significant association was found between expression of nm23‐H1 protein and prognosis (p < 0.01). Expression of nm23‐H1 protein indicated favorable prognosis, suggesting that the absence of nm23‐H1 protein expression was significantly associated with lymph‐node metastasis, recurrence and distant metastasis in NPC. Expression of the nm23‐H1 gene may be valuable for assessing the prognosis of NPC. Int. J. Cancer (Pred. Oncol.) 79:596–600, 1998.

Nasopharyngeal extranodal NK/T-Cell lymphoma, nasal type: Retrospective study of 18 consecutive cases in Guangzhou, China

Objective: The aim of this study was to investigate the frequency and clinicopathologic features of nasopharyngeal extranodal NK/T-cell lymphoma, nasal type (NKTCL), as well as DNA sequence variation of Epstein—Barr virus (EBV) in neoplastic cells harboring in NKTCLs from Guangzhou district. Materials and methods: The clinical data of 18 unselected consecutive nasopharyngeal NKTCLs in one institution were reviewed retrospectively. Immunohistochemical staining and EBV-encoded RNAs (EBERs) in situ hybridization were applied. DNA extraction, polymerase chain reaction (PCR), nested PCR, and sequencing for analyzing the C-terminal and N-terminal regions of LMP1 gene as well as BamHI F fragment of EBV were applied in 16 available samples. Results: NKTCLs accounted for 69.2% (18/26) of nasopharyngeal T- and NK-cell lineage non-Hodgkin lymphomas. In all, 10 out of 18 patients (55.56%) had cervical lymph node(s) involvement. The serum anti-EBV antibody level was elevated (VCA-IgA titer ≥1:40) in 6 of 12 available patients. Two major immunophenotypic subtypes, namely, TIA-1+/EBERs+/CD56+ (10 cases) and TIA-1+/EBERs+/CD56- (8 cases) could be recognized. Genotyping analysis revealed that 10 out of 13 cases (76.9%) of NKTCL were harbored with del-LMP1 [del-LMP1 (Gly335) variant 7 cases, del-LMP1 (Asp335) variant 3 cases]. XhoI-loss was shown in 8/11 cases (72.73%). BamHI “f” variant of Bam F fragment was shown only in 4/14 cases (28.57%).The most common combination was del-LMP1 (Gly335)/ XhoI-loss/F (6/9, 66.7%). Conclusions: The majority of nasopharyngeal T- and NK-cell lymphomas are NKTCL in Guangzhou district. The patients often have involvement of cervical lymph node(s) and an elevated level of serum anti-EBV antibodies. The CD56 expression rate seems lower than that found in sinonasal NKTCL. The most common EBV variant harboring in nasopharyngeal NKTCL seems somewhat different from that harboring in nasopharyngeal carcinoma in Guangzhou.

Mast cells in adjacent normal colon mucosa rather than those in invasive margin are related to progression of colon cancer

ObjectiveMast cells (MC) reside in the mucosa of the digestive tract as the first line against bacteria and toxins. Clinical evidence has implied that the infiltration of mast cells in colorectal cancers is related to malignant phenotypes and a poor prognosis. This study compared the role of mast cells in adjacent normal colon mucosa and in the invasive margin during the progression of colon cancer.MethodsSpecimens were obtained from 39 patients with colon adenomas and 155 patients with colon cancers treated at the Sun Yat-sen University Cancer Center between January 1999 and July 2004. The density of mast cells was scored by an immunohistochemical assay. The pattern of mast cell distribution and its relationship with clinicopathologic parameters and 5-year survival were analyzed.ResultsThe majority of mast cells were located in the adjacent normal colon mucosa, followed by the invasive margin and least in the cancer stroma. Mast cell count in adjacent normal colon mucosa (MCCadjacent) was associated with pathologic classification, distant metastases and hepatic metastases, although it was not a prognostic factor. In contrast, mast cell count in the invasive margin (MCCinvasive) was associated with neither the clinicopathlogic parameters nor overall survival.ConclusionMast cells in the adjacent normal colon mucosa were related to the progression of colon cancer, suggesting that mast cells might modulate tumor progression via a long-distance mechanism.