Centromere Protein H Expression Research Articles

CENPH Inhibits Rapamycin Sensitivity by Regulating GOLPH3-dependent mTOR Signaling Pathway in Colorectal Cancer.

Background: Centromere protein H (CENPH) is known as a fundamental component of the active centromere complex, and its overexpression is correlated with poor prognosis in various solid tumors. mTOR inhibitor rapamycin has been shown to possess antitumor activity, as well as prevent intestinal tumorigenesis. However, the prognostic value of CENPH in colorectal cancer (CRC) and the role of CENPH in rapamycin sensitivity remain unknown.Materials and methods: The effect of CENPH on the cell proliferation, clonogenicity, and cell response to rapamycin in CRC were evaluated by MTT and/or colony formation assays. For the underlying mechanisms, the interaction between CENPH and GOLPH3 were detected by co-immunoprecipitation, GST pull-down, and His-tag pull-down assays, as well as the laser scanning confocal microscopy. The status of kinases in mTOR signaling was determined by Western blot. Finally, the clinical significance of CENPH was analyzed using public CRC datasets with CENPH transcripts and clinical information.Results: CENPH inhibited CRC malignant phenotypes, conferred reduced sensitivity to rapamycin, and attenuated both mTORC1 and mTORC2 in mTOR signaling pathway through the interaction with golgi phosphoprotein 3 (GOLPH3), which has been identified as a potential oncogene and modulates the response to rapamycin. Moreover, elevated levels of CENPH were detected in CRC tissues, compared with normal colorectal tissues. High levels of CENPH expression gradually decreased according to CRC tumor stages. Patients with high CENPH expression had favorable survival.Conclusions: Our results suggest that CENPH inhibits rapamycin sensitivity by regulating GOLPH3 dependent mTOR pathway. High CENPH expression is associated with better prognosis in CRC patients. Taken together, CENPH may serve as a potential predictor for rapamycin sensitivity and therapeutic target for CRC patients.

Upregulation of centromere protein H is associated with progression of renal cell carcinoma

Centromere protein H (CENPH), one of the essential component of active kinetochore, plays an important role in carcinogenesis of many cancer types. However, its expression signature and prognostic significance of renal cell carcinoma (RCC) are unclear. In the present study, we concluded that the expression of CENPH was prominently upregulated in RCC specimens and three RCC cell lines (ACHN, 786-O and A704). Immunohistochemical analysis revealed that RCCs exhibited higher levels of CENPH expression than normal renal tissues in paraffin-embedded archival specimens. Further statistical analysis suggested the upregulation of CENPH was positively correlated with the Fuhrman grade (P = 0.001), distant metastasis (P = 0.024) and clinical stage (P = 0.014). In addition, the CENPH served as an independent predictor of overall survival of RCC patients in multivariate analysis (P = 0.018). Furthermore, our in vitro assays of RCC cell lines indicated that knockdown of CENPH reduced cell proliferation, inhibited cell growth, and increased cell apoptosis. In conclusion, our data suggest that CENPH is a novel molecule involved in RCC progression, which provides a potential biomarker and therapeutic target.

Expression of centromere protein H and its correlation with clinicopathological features and prognosis in gastric cancer

Objective To explore the expression of centromere protein H (CENP-H) and its correlation with clinicopathological features of gastric cancer (GC).Methods The expression level of CENP-H in 9 caes of gastric cancer tissue and corresponding normal mucosal tissue was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting.CENP-H expression levels in specimens from 166 patients with GC and nonneoplastic mucosal tissue were determined by immunohistochemical staining.The correlations between CENP-H with clinicopathologic features and prognosis was analyzed.Results It was dearly showed that the mRNA and protein expression of CENP-H was elevated in tumor tissue samples as compared with that in nonneoplastic mucosal tissue (P ＜ 0.05).The CENP-H expression was positive in 85 of 166 cases of GC,and negative in all nonneoplastic mucosal tissues,showing a significant correlation with tumour size (P ＜ 0.05),depth of infiltration (P ＜ 0.01),lymph node metastasis (P ＜ 0.01) and TNM staging (P ＜0.01) of GC,as well as clinical prognosis (P ＜0.01).Log-rank test indicated that 5-year survival rate of patients with high CENP-H expression was significantly lower than that of patients with low CENP-H expression.Multivariate Cox regression analysis confirmed that CENP-H (P ＜0.05),lymph node metastasis (P ＜ 0.01) and TNM staging (P ＜ 0.01) were independent prognostic predictors for patients with GC.Conclusion Our data suggested that CENP-H might play a role as a cancer gene and that its high expression indicated a poor prognosis in GC. Key words: Centromere protein H; Gastric Carcinoma; Prognosis

Overexpression of CENP-H as a novel prognostic biomarker for human hepatocellular carcinoma progression and patient survival

Centromere protein H (CENP-H) has been shown to be significantly upregulated in many types of cancers and is associated with disrupted cell cycle regulation, cell proliferation and genetic instability. The aim of the present study was to explore the expression and localization of CENP-H in hepatocellular carcinoma (HCC) and determine whether its overexpression is a prognostic biomarker for HCC. Reverse transcription-polymerase chain reaction (pcr), real-time qPCR and western blotting were used to compare CENP-H expression at the mRNA and protein levels in HCC samples and corresponding adjacent non-cancerous samples. CENP-H protein levels were determined in 60 paired paraffin-embedded HCC tissues using immunohistochemistry (IHC), and the correlation with clinicopathological features and patient prognosis was analyzed. In addition, an immunofluorescence assay was performed to test the expression and localization of CENP-H protein in HCC cells. Results showed that levels of CENP-H mRNA and protein were higher in HCC samples than in the corresponding adjacent non-cancerous samples. In 60 paired paraffin-embedded tissues, CENP-H was upregulated in the HCC samples (38/60, 63.3%) relative to the adjacent non-cancerous samples (21/60, 35%, P=0.003), and a higher level of upregulation was associated with tumor size (P=0.032); higher histological grade (P=0.001); more advanced TNM stage (P=0.002) and Chinese clinical stage (P=0.008); and poorer prognosis. In addition, consistent with the results of IHC, the immunofluorescence assay showed that CENP-H was localized in the nucleus of Hep3B cells. CENP-H was overexpressed in HCC, and its level of upregulation was an independent prognostic indicator, suggesting that CENP-H may be an effective therapeutic strategy for the treatment of HCC.

Clinical Significance of CENP-H Expression in Uterine Cervical Cancer.

ObjectiveThis work aims to investigate the expression pattern and clinicopathologic significance of centromere protein H (CENP-H) in uterine cervical cancer (UCC).MethodsThe level of CENP-H expression in the paraffin sections of 62 UCC cases was determined by the SP immunohistochemical method, with complete clinicopathologic data in all cases. Statistical analysis was conducted to evaluate the prognostic and diagnostic significance of CENP-H using SPSS13.0 software package.ResultsImmunohistochemical assay showed strong CENP-H expression in 61.29% (38/62) of the paraffin-embedded cervical cancer tissues. Statistical analysis revealed a strong correlation between the CENP-H expression and the clinical classification (P=0.038) of the cervical carcinoma. The expression increased with rise of the stages. The analysis of Cox proportional hazards regression model suggested that CENP-H expression (P=0.002) and tumor stage (P=0.001) were independent prognostic markers for the survival of UCC patients. The survival analysis showed that the survival rate was significantly lower in patients with high expression of CENP-H than in those with low expression of CENP-H (P=0.001).ConclusionsCENP-H is likely to be a valuable marker for carcinogenesis and progression of UCC. It might be used as the important diagnostic and prognostic marker for cervical carcinoma patients, especially for those at early stage.

Role of Centromere Protein H and Ki67 in Relapse-free Survival of Patients after Primary Surgery for Hypopharyngeal Cancer

Centromere protein H (CENP-H) and Ki67 are overexpressed in some malignancies, but whether they are predictors of survival after primary resection for hypopharyngeal squamous cell carcinoma (HSCC) remains unknown. We assessed immunohistochemical expression of CENP-H and Ki67 in 112 HSCC specimens collected between March 2003 and March 2005 for analysis by clinical characteristics. The Kaplan-Meier method was used to analyze relapse-free survival and logistic multivariate regression to determine risk factors of relapse-free survival. Cholecystokinin octapeptide assays and flow cytometry were used to examine cell proliferation and apoptosis after siRNA inhibition of CENP-H in HSCC cells. Overall, 50 (44.6%) HSCC specimens showed upregulated CENP-H expression and 69 (61.6%) upregulated Ki67. An increased CENP-H protein level was associated with advanced cancer stage and alcohol history (P=0.012 and P=0.048, respectively) but an increased Ki67 protein level only with advanced cancer stage (P=0.021). Increased CENP-H or Ki67 were associated with short relapse-free survival (P<0.001 or P=0.009, respectively) and were independent predictors of relapse-free survival (P=0.001 and P=0.018, respectively). siRNA knockdown of CENP-H mRNA inhibited cell proliferation and promoted cancer cell apoptosis in vitro. Upregulated CENP-H and Ki67 levels are significantly associated with short relapse-free survival in HSCC. These factors may be predictors of a relapsing phenotype in HSSC cases.

Overexpression of centromere protein H is significantly associated with breast cancer progression and overall patient survival

Breast cancer is one of the leading causes of cancer death worldwide. This study aimed to analyze the expression of centromere protein H (CENP-H) in breast cancer and to correlate it with clinicopathologic data, including patient survival. Using reverse transcription-polymerase chain reaction and Western blotting to detect the expression of CENP-H in normal mammary epithelial cells, immortalized mammary epithelial cell lines, and breast cancer cell lines, we observed that the mRNA and protein levels of CENP-H were higher in breast cancer cell lines and in immortalized mammary epithelial cells than in normal mammary epithelial cells. We next examined CENP-H expression in 307 paraffin-embedded archived samples of clinicopathologically characterized breast cancer using immunohistochemistry, and detected high CENP-H expression in 134 (43.6%) samples. Statistical analysis showed that CENP-H expression was related with clinical stage (P = 0.001), T classification (P = 0.032), N classification (P = 0.018), and Ki-67 (P < 0.001). Patients with high CENP-H expression had short overall survival. Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient survival. Our results suggest that CENP-H protein is a valuable marker of breast cancer progression and prognosis.

Centromere protein H is a novel prognostic marker for human nonsmall cell lung cancer progression and overall patient survival

Lung cancer is 1 of the leading causes of cancer death worldwide, and the high mortality from this disease is caused mainly by the lack of efficient diagnostic strategies for early-stage lung cancer. The objective of the current study was to investigate the expression pattern and clinicopathologic significance of centromere protein H (CENP-H) in patients with nonsmall cell lung cancer (NSCLC). The expression profile of CENP-H in normal lung epithelial cells, NSCLC cell lines, NSCLC tissues, and adjacent noncancerous lung tissues were detected by reverse transcription-polymerase chain reaction (RT-PCR), real-time RT-PCR, and Western blot analysis. The expression level of CENP-H in 223 NSCLC tissues was measured by immunohistochemistry staining. Statistical analysis was performed to evaluate the clinicopathologic significance of CENP-H. The expression level of CENP-H was much higher in cancer cell lines and lung cancer tissues than that in normal cells and adjacent noncancerous lung tissues, respectively. Immunohistochemical analysis revealed positive CENP-H expression in 118 of 223 NSCLC tissues (52.9%). Statistical analysis revealed that CENP-H expression was correlated strongly with clinical stage (P=.018), tumor classification (P=.03), and Ki-67 expression (P < .001). Patients with lower CENP-H expression had better overall survival than patients with higher CENP-H expression. Further analysis suggested that CENP-H could predict prognosis only in patients with early-stage disease. Multivariate analysis suggested that CENP-H expression was an independent prognostic marker for survival in patients with NSCLC. The current results demonstrated that high CENP-H protein expression was related to poor outcome in patients with NSCLC. CENP-H may be used as a prognostic biomarker for patients lung patients, especially those with early-stage NSCLC.

Prognostic relevance of Centromere protein H expression in esophageal carcinoma

BackgroundMany kinetochore proteins have been shown to be associated with human cancers. The aim of the present study was to clarify the expression of Centromere protein H (CENP-H), one of the fundamental components of the human active kinetochore, in esophageal carcinoma and its correlation with clinicopathological features.MethodsWe examined the expression of CENP-H in immortalized esophageal epithelial cells as well as in esophageal carcinoma cells, and in 12 cases of esophageal carcinoma tissues and the paired normal esophageal tissues by RT-PCR and Western blot analysis. In addition, we analyzed CENP-H protein expression in 177 clinicopathologically characterized esophageal carcinoma cases by immunohistochemistry. Statistical analyses were applied to test for prognostic and diagnostic associations.ResultsThe level of CENP-H mRNA and protein were higher in the immortalized cells, cancer cell lines and most cancer tissues than in normal control tissues. Immunohistochemistry showed that CENP-H was expressed in 127 of 171 ESCC cases (74.3%) and in 3 of 6 esophageal adenocarcinoma cases (50%). Statistical analysis of ESCC cases showed that there was a significant difference of CENP-H expression in patients categorized according to gender (P = 0.013), stage (P = 0.023) and T classification (P = 0.019). Patients with lower CENP-H expression had longer overall survival time than those with higher CENP-H expression. Multivariate analysis suggested that CENP-H expression was an independent prognostic marker for esophageal carcinoma patients. A prognostic value of CENP-H was also found in the subgroup of T3~T4 and N0 tumor classification.ConclusionOur results suggest that CENP-H protein is a valuable marker of esophageal carcinoma progression. CENP-H might be used as a valuable prognostic marker for esophageal carcinoma patients.

Centromere Protein H Is a Novel Prognostic Marker for Nasopharyngeal Carcinoma Progression and Overall Patient Survival

The aim of the present study was to analyze the expression of Centromere protein H (CENP-H), one of the fundamental components of the human active kinetochore, in nasopharyngeal carcinoma (NPC) and to correlate it with clinicopathologic data, including patient survival. Using reverse transcription-PCR and Western blot, we detected the expression of CENP-H in normal nasopharyngeal epithelial cells, immortalized nasopharyngeal epithelial cell lines, and NPC cell lines. Using immunohistochemistry, we analyzed CENP-H protein expression in 160 clinicopathologically characterized NPC cases. Statistical analyses were applied to test for prognostic and diagnostic associations. Reverse transcription-PCR and Western blot showed that the expression level of CENP-H was higher in NPC cell lines and in immortalized nasopharyngeal epithelial cells than in the normal nasopharyngeal epithelial cell line at both transcriptional and translational levels. By immunohistochemical analysis, we found that 76 of 160 (47.5%) paraffin-embedded archival NPC biopsies showed high expression of CENP-H. Statistical analysis showed that there was a significant difference of CENP-H expression in patients categorized according to clinical stage (P = 0.024) and T classification (P = 0.027). Patients with higher CENP-H expression had shorter overall survival time, whereas patients with lower CENP-H expression had better survival. A prognostic value of CENP-H was also found of the subgroup of N(0)-N(1) tumor classification. Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient's survival. Our results suggest that CENP-H protein is a valuable marker of NPC progression. High CENP-H expression is associated with poor overall survival in NPC patients.

Increased expression of CENP-H gene in human oral squamous cell carcinomas harboring high-proliferative activity

We examined the expression of Centromere protein H (CENP-H) mRNA in 38 oral squamous cell carcinomas (SCCs), 2 epithelial dysplasias and 5 normal gingivae using the real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). The mean expression level of CENP-H mRNA was higher in oral SCCs (0.11+/-0.08) than epithelial dysplasias (0.03+/-0.01) and normal gingivae (0.027+/-0.01). The expression level of CENP-H mRNA was significantly higher in oral SCCs than normal gingivae (Mann-Whitney U test, P=0.005). We also found a significant association between the level of CENP-H mRNA expression and clinical stage in oral SCCs (Mann-Whitney U test, P=0.04). We next studied the expression of CENP-H in 17 oral SCCs immunohistochemically. A significant correlation between the expression levels of CENP-H protein and the Ki-67 labeling index was found (Mann-Whitney U test, P=0.005). These results indicate that human CENP-H is closely linked to the increased or abnormal cell proliferation in malignant conditions.