OBJECTIVE: Human U three protein 14a (hUTP14a) facilitates tumorigenesis through promoting p53 and Rb degradation as well as enhancing c-Myc oncogenic activity. Moreover, hUTP14a expression is up-regulated in human hepatocellular cancer and colorectal cancer tissues. In this study, the expression of hUTP14a in non-small cell lung cancer (NSCLC) tissues was evaluated by immunohistochemistry staining (IHC). The relationship between hUTP14a expression levels and the clinical characteristics of the NSCLC patients were analyzed. METHODS: Lung cancer tissues and the adjacent non-cancerous tissues were collected from 123 cases of NSCLC patients including 53 cases of squamous cell carcinoma (SCC) and 70 cases of adenocarcinoma (ADC), who had accepted surgical resection at Peking University Third Hospital from May 2003 to April 2006. The expression level of hUTP14a was determined by IHC in human NSCLC tissues and the adjacent non-cancerous tissues. The associations between hUTP14a expression and the clinical pathological variables including gender, age, tumor size, histological type, differentiation degree and clinical pathological stage were analyzed using the Pearson's χ² test. RESULTS: The expression rate of hUTP14a in NSCLC tissues was significantly higher than that in the non-cancerous tissues (37.4% vs. 0, P<0.001). The expressions of hUTP14a in lung ADC and SCC were 48.6% and 20.6%, respectively. The expression rate of hUTP14a in both lung ADC and SCC was significantly higher than that in the adjacent non-cancerous tissues (P<0.001). In addition, the expression rate of hUTP14a in lung ADC was significantly higher than that in SCC (χ²=8.66, P=0.003). Furthermore, the expression rate of hUTP14a in the late pTNM stage of SCC was significantly higher than that in the early pTNM stage of SCC while hUTP14a expression level was not associated with pTNM stage of ADC. No correlation was found between hUTP14a expression and the other clinical pathologic features of the patients. CONCLUSION Expression of hUTP14a was up-regulated in NSCLC tissues and was correlated with pTNM stage of SCC, suggesting that hUTP14a might possess a potential as a candidate marker for the early diagnosis screening of NSCLC.
Adenocarcinoma ; Carcinoma, Non-Small-Cell Lung ; Carcinoma, Squamous Cell ; Humans ; Lung Neoplasms ; Prognosis ; Ribonucleoproteins, Small Nucleolar/metabolism*

BACKGROUND: Recurrent somatic SET-binding protein 1 (SETBP1) and splicing pathway gene mutations have recently been found in atypical chronic myeloid leukemia and other hematologic malignancies. These mutations have been comprehensively analyzed in adult AML, but not in childhood AML. We investigated possible alteration of the SETBP1, splicing factor 3B subunit 1 (SF3B1), U2 small nuclear RNA auxiliary factor 1 (U2AF1), and serine/arginine-rich splicing factor 2 (SRSF2) genes in childhood AML. METHODS: Cytogenetic and molecular analyses were performed to reveal chromosomal and genetic alterations. Sequence alterations in the SETBP1, SF3B1, U2AF1, and SRSF2 genes were examined by using direct sequencing in a cohort of 53 childhood AML patients. RESULTS: Childhood AML patients did not harbor any recurrent SETBP1 gene mutations, although our study did identify a synonymous mutation in one patient. None of the previously reported aberrations in the mutational hotspot of SF3B1, U2AF1, and SRSF2 were identified in any of the 53 patients. CONCLUSIONS: Alterations of the SETBP1 gene or SF3B1, U2AF1, and SRSF2 genes are not common genetic events in childhood AML, implying that the mutations are unlikely to exert a driver effect in myeloid leukemogenesis during childhood.
Adolescent ; Carrier Proteins/*genetics ; Child ; Child, Preschool ; Cohort Studies ; Cytogenetic Analysis ; DNA Mutational Analysis ; Female ; Gene Frequency ; Genotype ; Humans ; Infant ; Leukemia, Myeloid, Acute/*genetics/pathology ; Male ; Nuclear Proteins/*genetics ; Phosphoproteins/*genetics ; Polymorphism, Single Nucleotide ; RNA Splicing ; Ribonucleoprotein, U2 Small Nuclear/*genetics ; Ribonucleoproteins/*genetics

PURPOSE: Prader-Willi Syndrome(PWS) is caused by absence of paternal contributions of the chromosome region 15q11-q13. To detact this region, high resolutional cytogenetic analysis, FISH with probe at PWS critical region or microsatellite polymorphism can be used. The gene for the small nuclear ribonucleoprotein polypeptide N(SNRPN) is not expressed in patients with PWS. We conducted molecular analysis with RT-PCR with SNRPN primers to find out more useful diagnostic tool in PWS. METHODS: Four patients with obesity and other characteristics of PWS were studied. The exprssion of SNRPN and control gene were studed by RT-PCR from peripheral lymphocytes. RESULTS :The SNRPN expression in reverse transcribed RNA from blood were easily detected in normal control but not in patients with suspected Parder-Willi Syndrome. CONCLUSION: We conclude that SNRPN expression study is a useful diagnostic method for detection of Prader-Willi Syndrome.
Cytogenetic Analysis ; Humans ; Lymphocytes ; Microsatellite Repeats ; Obesity ; Pathology, Molecular* ; Prader-Willi Syndrome* ; Ribonucleoproteins, Small Nuclear ; RNA ; snRNP Core Proteins*

BACKGROUND: The histogenesis and interrelationship of the various types of germ cell tumors (GCTs) have been proposed. Dysgerminoma/seminoma (D/S) is a primitive GCT that has not acquired the potential for further differentiation, whereas other types of GCTs are in a dynamic process of differentiation towards a somatic or extraembryonal direction. A primordial germ cell giving rise to a GCT undergoes a developmentally regulated erasure and resetting of imprinted genes, but changes in the imprinting pattern in GCTs as the tumor differentiates have not been well defined. We aimed to investigate the changes of the SNRPN methylation pattern between the germinomas and non-germinomatous GCTs, as compared with the somatic methylation pattern. METHODS: We used formalin-fixed paraffin-embedded tissue sections of 97 GCTs (18 Ds, 21 Ss, 17 yolk sac tumors (YSTs), 19 immature teratomas, and 22 mature teratomas). DNA methylation was evaluated after bisulfite modification, PCR amplification, and restriction enzyme digestion. RESULTS: The SNRPN methylation pattern was changed in 53/74 (71.6%) of GCTs as non-somatic patterns. There were significant differences in the methylation pattern between the germinomas and non-germinomatous GCTs, the GCTs being frequently hypo- methylated in Ds/Ss (73.3%), in contrast to the frequent hypermethylation seen in the YSTs and teratomas (47.7%, p<0.05). CONCLUSIONS: The methylation status of an imprinting gene may be involved in the mechanism causing cellular differentiation and tumorigenesis of GCTs.
Carcinogenesis ; Digestion ; DNA Methylation ; Endodermal Sinus Tumor ; Genomic Imprinting ; Germ Cells* ; Germinoma ; Humans* ; Methylation* ; Neoplasms, Germ Cell and Embryonal* ; Polymerase Chain Reaction ; Ribonucleoproteins, Small Nuclear* ; snRNP Core Proteins ; Teratoma

Telomerase is a ribonucleoprotein that synthesizes telomere repeats. It has been reported that activation of telomerase was associtated with immortalization, proliferative activity and carcinogenesis. Recently, telomerase activity has been extensively studied in many kinds of malignant tumors for clinical diagnostic and/or prognostic utilities. In neuroblastoma, breast carcinoma,gastric carcinoma, non-small cell lung carcinoma, close relationship has been reported between high telomerase activity and lymph node metastasis, tumor aggressiveness and poor prognosis. The purpose of this study is to to investigate the clinical implication of telomerase activity assay as an adjunctive factor in decision-making on neck node management, speedy pre-operative judging on histologic malignancy grading. Thus we performed semi-quantitative assay of telomerase activity using Telomerase PCR ELISA kit(Boeringer Manheim , Germany) and evaluated correlation between telomerase activity and tumor size, neck node metastasis, Anneroth malignancy score and influence of pre-operative chemotherapy on its activity in 27 cases of oral squamous cell carcinomas and 18 cases of normal oral epithelium. Also, correlation between telomerase activities and PCNA indices was evaluated. The results were obtained as follows: 1. The telomerase activities were detected in 24 specimens out of 27 oral squamous cell carcinoma specimens (88.9%) and in 5 specimens out of 18 normal oral epithelium specimens (27.8%). The mean value of telomerase activities was 0.9793+/-0.3428 in 24 oral squamous cell carcinoma specimens and 0.4855+/-0.1117 in 5 normal oral epithelium specimens. The positivity rate and mean value of telomerase activities in oral squamous cell carcinoma specimens were significantly higher than those of normal oral epithelium specimens (p<0.05). 2. There was no significant correlation between total Anneroth malignancy score and telomerase activity (p>0.05), but points of mitosis index and depth of invasion were significantly correlated with telomerase activities (p<0.05). 3. The positive immunohistochemical staining for PCNA(proliferating cell nuclear antigen) was observed in 26 specimens out of 27 oral squamous cell carcinoma specimens and mean value of PCNA indices of 26 specimens was 53.67+/-26.46. PCNA indices were significantly correlated with telomerase activities (p<0.05). 4. The mean value of telomerase activities was significantly higher in pathologic T3/T4 group than in T1/T2 group (p<0.01). There was no significant difference of mean value of telomerase activities between pathologic neck node positive group and negative group (p> 0.05). Pre-operative chemotherapy significantly lowered the telomerase activities (p<0.05). The above results suggested telomerase activity could be used as diagnostic marker and adjunctive parameter for judging on histologic malignancy in oral squamous cell carcinoma.
Breast ; Carcinogenesis ; Carcinoma, Non-Small-Cell Lung ; Carcinoma, Squamous Cell ; Drug Therapy ; Enzyme-Linked Immunosorbent Assay ; Epithelium ; Lymph Nodes ; Mitosis ; Neck ; Neoplasm Metastasis ; Neuroblastoma ; Polymerase Chain Reaction ; Prognosis ; Proliferating Cell Nuclear Antigen ; Ribonucleoproteins ; Telomerase* ; Telomere

OBJECTIVE: The regulation of apoptosis is influenced by various gene products including Bcl-2, which has been known to be anti-apoptotic. In addition, telomerase, a ribonucleoprotein that synthesizes telomeres, has been detected in many human neoplasms. In the current study, we present the anti-apoptotic effect by Bcl-2 expression and telomerase activity in the human brain tumors. METHODS: A total of 76 cases of surgically resected brain tumors were studied. Telomerase activity was detected by the telomeric repeat amplification protocol assay and Bcl-2 protein was examined by the Western blot analysis. Apoptosis of the specimens was also detected by DNA fragmentation analysis. RESULTS: Telomerase activity and apoptosis were detected in 65.8%(50 of 76) and 21.1%(16 of 76) respectively. Telomerase activity was correlated to apoptosis inversely(p<0.05). Bcl-2 was also expressed in 23.7%(18 of 76) and apoptosis detected in 11.1%(2 of 18) with Bcl-2. In 18 cases with Bcl-2, telomerase activity was expressed in 77.8%(14 of 18) and apoptosis was not induced in 85.7%(12 of 14) with telomerase activity. However, apoptosis was not detected in 4 cases with Bcl-2 and negative telomerase activity. Their difference of anti-apoptotic numbers between two groups was significant(p<0.05). In 36 cases with negative Bcl-2 and positive telomerase activity, apoptosis was not detected in 72.2%(26 of 36). In 22 cases without Bcl-2 and telomerase activity, apoptosis was not detected in 81.8%(18 of 22). Their difference between two groups was not significant statistically. CONCLUSION: Our results suggest that apoptosis may be modulated by telomerase activity of the human brain tumors with Bcl-2. And also, we may expect that apoptosis can be induced effectively by the inhibition of telomerase activity and Bcl-2 in the brain tumors.
Apoptosis* ; Blotting, Western ; Brain Neoplasms* ; Brain* ; DNA Fragmentation ; Humans ; Ribonucleoproteins ; Telomerase* ; Telomere

Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric DNA onto the ends of chromosomes. thereby preventing the replication-dependent shortening of these ends. Telomerase activity is detected in a wide range of cancers of various tissues, and its expression may be a critical step in tumor progression. Our objective was to determine if detection of telomerase activity may be an indicator for diagnosis of breast cancer and any association between telomerase activity and prognostic factors of breast cancer. Using a polymerase chain reaction-based telomerase activity assay, we examined telomerase activity in 30 breast cancer specimens (2 ductal carcinoma in situ, 28 invasive ductal carcinoma), 25 benign lesions (14 fibroadenomas, 11 fibrocystic diseases) and 24 normal breast tissues (13 adjacent to malignancy, 11 adjacent to benign lesion). Among surgically resected samples, telomerase activity was detected in 23 (77%) of 30 breast cancers. While telomerase activity was not detected in any of 11 specimens of fibrocystic disease and 11 adjacent normal tissues to benign lesion, surprisingly low levels of telomerase activity were detected in 5 (36%) of 14 fiboadenomas and 1 (7%) of 13 adjacent normal tissues to malignancy. There was no significant difference in expression of telomerase among prognostic factors of breast cancer. In summary, telomerase activity in breast cancer may be useful in diagnosis of breast cancer. We found no correlation between telomerase activity and stage, tumor size or LN status. Mechanisms of telomerase expression are still under investigation; therefore, the significance of telomerase expression in malignant tumors and their progression remains to be determined.
Breast Neoplasms* ; Breast* ; Carcinoma, Intraductal, Noninfiltrating ; Diagnosis ; DNA ; Fibroadenoma ; Humans* ; Ribonucleoproteins ; Telomerase*

PURPOSE: Telomerase is a ribonucleoprotein involved in maintaining the telomere length in stem, and immortal or actively dividing, cells. There is controversy relating to the correlation between the telomerase activity and the clinicopathological characteristics of renal cell carcinomas. The relationships between the telomerase activity and the clinicopathological characteristics of renal cell carcinomas were evaluated. MATERIALS AND METHODS: Twenty-seven renal cell carcinoma tissues, and 22 normal renal tissues around renal cell carcinoma tissues, were aseptically obtained from a radical or partial nephrectomy in 27 cases with a renal cell carcinoma. The telomerase activity was analyzed using a PCR-based telomeric repeat amplification protocol (TRAP)-ELISA method. The telomerase activity was compared with the clinicopathological characteristics, such as sex, age of patient, histologic type, stage, grade, size and metastasis, of the renal cell carcinomas. RESULTS: Telomerase activity was detected in 24 of the 27 renal cell carcinoma tissues (88.9%), but not in all of the normal renal tissues. There was no statistical correlation between the telomerase activity and the clinicopathological features. CONCLUSIONS: These findings indicate that the telomerase activity may play a role in the carcinogenesis of renal cell carcinomas, but there is no relationship between the telomerase activity and the clinicopathological characteristics of renal cell carcinomas.
Carcinogenesis ; Carcinoma, Renal Cell* ; Humans ; Neoplasm Metastasis ; Nephrectomy ; Ribonucleoproteins ; Telomerase* ; Telomere

BACKGROUND: Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric DNA onto the ends of chromosomes, thereby preventing the replication-dependent shortening of those ends. Telomerase activity is detected in a wide range of cancers of various tissues, and its expression may be a critical step in tumor progression. Our objective was to determine if detection of telomerase activity may be an indicator for diagnosis of breast cancer and if any association exists between telomerase activity and prognostic factors of breast cancer. METHODS: Using a polymerase chain reaction-based telomerase activity assay, we examined telomerase activity in 30 breast cancer specimens (2 ductal carcinoma in situ, 28 invasive ductal carcinoma), 25 benign lesions (14 fibroadenomas, 11 fibrocystic diseases), and 24 normal breast tissues (13 adjacent to malignancy, 11 adjacent to benign lesion). RESULTS: Among surgically resected samples, telomerase activity was detected in 23 (77%) of 30 breast cancers. While telomerase activity was not detected in any of the 11 specimens of fibrocystic disease and the 11 normal tissues adjacent to benign lesion, surprisingly low levels of telomerase activity were detected in 5 (36%) of the 14 fibroadenomas and 1 (7%) of the 13 normal tissues adjacent to malignancy. There was no significant difference in expression of telomerase among prognostic factors of breast cancer. CONCLUSIONS: In summary, telomerase activity may be useful in the diagnosis of breast cancer. We found no correlation between telomerase activity and stage, tumor size, or LN status. Mechanisms of telomerase expression are still under investigation; therefore, the significance of telomerase expression in malignant tumors and their progression remains to be determined.
Breast Neoplasms* ; Breast* ; Carcinoma, Intraductal, Noninfiltrating ; Diagnosis ; DNA ; Fibroadenoma ; Humans* ; Ribonucleoproteins ; Telomerase*

BACKGROUND: Human telomerase is a ribonucleoprotein polymerase, which synthesizes telomeric repeat sequences, and human telomerase reverse transcriptase (hTERT) has been identified as the catalytic subunit, as well as the rate-limiting component, of telomerase. In this study, we attempted to identify the modulators of telomerase, and to determine the molecular mechanisms underlying cisplatin-induced apoptosis. METHODS: To determine the role of telomerase in cisplatin-induced apoptosis, we measured telomerase activity and analyzed apoptosis using PI and trypan blue staining. Also, we inhibited the caspase activations using Z-VAD-fmk to analyze the effects on expression of hTERT protein. Finally, we induced the transient co-expression of the Bcl-2 and Bak genes in HEK293 cells, and then, the telomerase activity and expression of hTERT were evaluated. RESULTS: In the Bcl-2-overexpressing HeLa cells, telomerase activity was more enhanced, and cell death was reduced to 40-50% that of the mock controls. This finding suggests that Bcl-2-induced telomerase activity exerts an antiapoptotic effect in cisplatin-induced death. As caspase activation was inhibited via Z-VAD-fmk, the hTERT protein was recovered in the mock controls, but not in the Bcl-2-overexpressing cells. This suggests that the expression of hTERT can be regulated by caspases, but Bcl-2 was located within the upstream pathway. Moreover, when the Bcl-2 and Bak genes were co-transfected into the HEK293, both telomerase activity and hTERT protein were prominently reduced. CONCLUSIONS: Bcl-2-induced telomerase activity inhibits cisplatin-induced apoptosis in HeLa cells, and can be regulated via both caspases and the interaction of Bcl-2 and Bak.
Apoptosis ; Caspases* ; Catalytic Domain ; Cell Death* ; Cisplatin ; HEK293 Cells ; HeLa Cells ; Humans* ; Ribonucleoproteins ; Telomerase* ; Trypan Blue