Circadian rhythm variation of the clock genes Per1 and cell cycle related genes in different stages of carcinogenesis of buccal mucosa in animal model / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and circadian rhythm variation of biological clock gene Per1 and cell cycle genes p53, CyclinD1, cyclin-dependent kinases (CDK1), CyclinB1 in different stages of carcinogenesis in buccal mucosa and its relationship with the development of buccal mucosa carcinoma.</p><p><b>METHODS</b>Ninety golden hamsters were housed under 12 hours light-12 hours dark cycles, and the model of buccal squamous cell carcinoma was established by using the dimethylbenzanthracene (DMBA) to smear the golden hamster buccal mucosa. Before the DMBA was used and after DMBA was used 6 weeks and 14 weeks respectively, the golden hamsters were sacrificed at 6 different time points (5 rats per time point) within 24 hour, including 4, 8, 12, 16, 20 and 24 hour after lights onset (HALO), and the normal buccal mucosa, precancerous lesions and cancer tissue were obtained, respectively. HE stained sections were prepared to observe the canceration of each tissue. Real time RT-PCR was used to detect the mRNA expression of Per1, p53, CyclinD1, CDK1 and CyclinB1, and a cosine analysis method was applied to determine the circadian rhythm variation of Per1, p53, CyclinD1, CDK1 and CyclinB1 mRNA expression, which were characterized by median, amplitude and acrophase.</p><p><b>RESULTS</b>The expression of Per1, p53, CDK1 and CyclinD1 mRNA in 6 different time points within 24 hours in the tissues of three different stages of carcinogenesis had circadian rhythm, respectively. However, the CyclinB1 mRNA was expressed with circadian rhythm just in normal and cancer tissue (P < 0.05), while in precancerous lesions the circadian rhythm was in disorder (P > 0.05). As the development of carcinoma, the median of Per1 and p53 mRNA expression were significantly decreased (P < 0.05), yet the median of CDK1, CyclinB1 and CyclinD1 mRNA expression were significantly increased (P < 0.05). The amplitude of Per1, p53 and CyclinD1 mRNA expression was significantly decreased as the development of carcinoma (P < 0.05), however the amplitude of CDK1 mRNA expression was significantly increased (P < 0.05). In addition, there was no significant difference in the amplitude of CyclinB1 mRNA expression. The time that the peak expression value of Per1 and CDK1 mRNA appeared (Acrophase) in precancerous lesions was remarkably earlier than that in normal tissues, but the acrophase of p53 and CyclinD1 mRNA expression was remarkably delayed. Moreover, the acrophase of CDK1 and CyclinB1 mRNA expression in cancer tissues was obviously earlier than that in normal tissues, yet there was no significant variation in acrophase of Per1, p53, CyclinD1 mRNA expression between normal tissues and cancer tissues.</p><p><b>CONCLUSIONS</b>The circadian rhythm of clock gene Per1 and cell cycle genes p53, CyclinD1, CDK1, CyclinB1 expression remarkably varied with the occurrence and development of carcinoma. Further research into the interaction between circadian and cell cycle of two cycle activity and relationship with the carcinogenesis may providenew ideas and methods of individual treatment and the mechanism of carcinogenesis.</p>

Association of glucocorticoid receptor polymorphisms with clinical and metabolic profiles in polycystic ovary syndrome

OBJECTIVES: We aimed to investigate whether glucocorticoid receptor gene polymorphisms are associated with clinical and metabolic profiles in patients with polycystic ovary syndrome. Polycystic ovary syndrome is a complex endocrine disease that affects 5-8% of women and may be associated with metabolic syndrome, which is a risk factor for cardiovascular disease. Cortisol action and dysregulation account for metabolic syndrome development in the general population. As glucocorticoid receptor gene (NR3C1) polymorphisms regulate cortisol sensitivity, we hypothesized that variants of this gene may be involved in the adverse metabolic profiles of patients with polycystic ovary syndrome. METHOD: Clinical, metabolic and hormonal profiles were evaluated in 97 patients with polycystic ovary syndrome who were diagnosed according to the Rotterdam criteria. The alleles of the glucocorticoid gene were genotyped. Association analyses were performed using the appropriate statistical tests. RESULTS: Obesity and metabolic syndrome were observed in 42.3% and 26.8% of patients, respectively. Body mass index was positively correlated with blood pressure, triglyceride, LDL-c, total cholesterol, glucose and insulin levels as well as HOMA-IR values and inversely correlated with HDL-c and SHBG levels. The BclI and A3669G variants were found in 24.7% and 13.4% of alleles, respectively. BclI carriers presented a lower frequency of insulin resistance compared with wild-type subjects. CONCLUSION: The BclI variant is associated with a lower frequency of insulin resistance in women with polycystic ovary syndrome. Glucocorticoid gene polymorphism screening during treatment of the syndrome may be useful for identifying subgroups of at-risk patients who would benefit the most from personalized treatment. .

Longer Survival in Patients with Breast Cancer with Cyclin D1 Over-Expression after Tumor Recurrence: Longer, but Occupied with Disease / 한국유방암학회지

PURPOSE: The effect of cyclin D1 overexpression on breast cancer outcomes and prognosis is controversial, even though amplification of the cyclin D1 gene, CCND1, has been shown to be associated with early relapse and poor prognosis. In this study, we examined the relationship between cyclin D1 overexpression and disease-specific survival (DSS). We also analyzed survival in patients who experienced recurrence. METHODS: We retrospectively analyzed data from patients diagnosed with ductal carcinoma between April 2005 and December 2010. We examined clinicopathologic factors associated with cyclin D1 overexpression and analyzed the influence of cyclin D1 on recurrence-free survival and DSS. RESULTS: We identified 236 patients diagnosed with primary breast cancer who completed all phases of their primary treatment. Cyclin D1 overexpression was significantly associated with longer DSS (5-year DSS, 89.9% in patients without cyclin D1 overexpression vs. 98.9% in patients with cyclin D1 overexpression; p=0.008). Multivariate analysis also found that patients with cyclin D1 overexpressing tumors had significantly longer disease-specific survival than patients whose tumors did not overexpress cyclin D1, with a hazard ratio for disease-specific mortality of 7.97 (1.17-54.22, p=0.034). However, in the group of patients who experienced recurrence, cyclin D1 overexpression was not significantly associated with recurrence-free survival. Cyclin D1 overexpression was significantly associated with increased survival after disease recurrence, indicating that cyclin D1 overexpression might be indicative of more indolent disease progression after metastasis. CONCLUSION: Cyclin D1 overexpression is associated with longer DSS, but not recurrence-free survival, in patients with breast cancer. Longer postrecurrence survival could explain the apparent inconsistency between DSS and recurrence-free survival. Patients with cyclin D1-overexpressing tumors survive longer, but with metastatic disease after recurrence. This information should spark the urgent development of tailored therapies to cure these patients.

Inhibitory effect of cyclin D1 antisense cDNA on human hepatocarcinoma cell line HepG2 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of antisense cDNA of cyclin D1 on the cyclin D1 gene expression and cell proliferation of human hepatocarcinoma HepG2 cells in vitro.</p><p><b>METHODS</b>Plasmids containing cyclin D1 antisense cDNA were constructed and transfected into HepG2 cells. Their effects on cell proliferation were examined by MTT method, RT-PCR, immunohistochemical means, and flow cytometry.</p><p><b>RESULTS</b>Cyclin D1 antisense cDNA significantly inhibited the growth of HepG2 cells. The inhibition peaked at 48 hour after transfection by MTT method. RT-PCR analysis showed that cyclin D1 antisense cDNA down-regulated cyclin D1 at the mRNA levels. Expression level of cyclin D1 protein was also decreased as shown by immunohistochemical studies. Cell-cycle analysis by flow cytometry showed that transfected HepG2 cells were arrested at the G1 phase of the cell cycle.</p><p><b>CONCLUSIONS</b>Our data suggest that cyclin D1 antisense cDNA could specifically inhibit the expression of cyclin D1 mRNA and protein and regulate cell cycle and cell proliferation of HepG2 cells. Cyclin D1 antisense cDNA may serve as a potential antitumor strategy in regulating cell-cyclin treating advanced HCCs.</p>

Cyclin D1 Expression in Primary Breast Carcinoma: Correlation with Estrogen Receptor Status and Other Clinicopathologic Parameters / 한국유방암학회지

PURPOSE: Cyclins are groups of proteins that play a role as a major regulator of the G1 restriction point promoting inactivation of the retinoblastoma protein. The cyclin D1 gene, CCND1, is amplified in approximately 20% of breast carcinomas and the protein is reportedly overexpressed in 60~80% of all cases. Cyclin D1 overexpression was strongly correlated to estrogen receptor positivity and better histologic grade in breast cancer. The aim of this study was to correlate cyclin D1 overexpression using a well characterized antibody with estrogen receptor status and other clincopathologic parameters. METHODS: From March 1989 to December 1994, 85 patients with primary breast carcinoma were the subject in this study. We analyzed cyclin D1 expression by immnohistochemical staining using cyclin D1 antibody, cells were considered positive according to distinct nuclear staining. The correlation between cyclin D1 expression was compared with important clinicopathologic parameters (tumor size, axillary lymph node status, p53 expression, c-erbB2 expression, histologic grade, estrogen receptor status). RESULTS: Cyclin D1 expression was detected in 37 cases (43.5%). Cyclin D1 expression was high in patients with tumors that expressed estrogen receptor (58.5% vs 26.5%, P=0.019). Cyclin D1 was mainly overexpressed in the histologic grade I and II (75.0%), as compared with 65.2% in cyclin D1 negative tumor, however there was no statistical significance (P=0.067). There were no significant correlation with tumor size, axillary lymph node status, p53 expression, or c-erbB2 expression (P>0.05). CONCLUSION: Cyclin D1 expression in estrogen receptor (ER) positive patients was significantly higher than that seen in ER negative patients. There was a negative correlation between cyclin D1 and tumor histologic grade, however it was not statistically significant. Tumor size, axillary lymph node status, p53 expression, and c-erbB2 expression were not correlated with cyclin D1.

Detection of IgH and Cyclin D1 gene Rearrangement with Interphase FISH in Multiple Myeloma / 대한진단검사의학회지

BACKGROUND: The t(11;14)(q13;q32) is known to be one of the most frequent chromosomal abnor-malities found in multiple myeloma (MM). However, studies on t(11;14) in MM have been problemat-ic due to the fact that MM cells proliferate poorly in vitro. The purpose of our study is to evaluate inci-dence, clinical, and hematologic findings of MM with IgH and cyclin D1 gene rearrangement and to investigate the usefulness of interphase FISH (fluorescence in situ hybridization). METHODS: The study group included 36 patients (23 newly diagnosed MM, 8 relapsed MM, 5 per-sistent MM after treatment) admitted to Mokdong and Gil Hospital from November 1998 to July 2002. Interphase FISH was performed with IGH/CCND1 dual color, dual fusion translocation probe (Vysis Inc, Downers Grove, IL USA), using bone marrow mononuclear cells. RESULTS: Incidence of IgH and cyclin D1 gene rearrangement by interphase FISH was 19%. One patient with normal karyotype and another patient without any metaphase cells showed IgH and cyclin D1 gene rearrangement with interphase FISH. The lambda light chain subtype was more frequently found in patients with rearrangement (4/5, 80%) than those without rearrangement (6/23, 26%) (P<0.05). No significant differences were found in other clinical and hematologic findings in the two groups. CONCLUSIONS: We suggest that MM with IgH and cyclin D1 gene rearrangement is associated with the expression of lambda light chain. Interphase FISH may be helpful in samples with normal karyotype or no metaphase cells for detection of gene rearrangement of MM.

Marcadores moleculares en pacientes con cáncer de cuello uterino tratadas con quimioterapia neoadyuvante y radioterapia / Molecular markers in patients with cervical cancer treated with induction chemotherapy and radiation therapy

Objetivos: En este estudio prospectivo de determinaron las modificaciones en la expresión y el valor predictivo de p53, p21 wafi/sdII/cipi, PCNA, hMLH1, hMSH2, Bcl'2 y TUNEL en pacientes con cáncer de cervix localmente avanzado tratadas con quimioterapia de inducción y radioterapia. Pacientes y métodos: Se obtuvieron muestras de 24 pacientes (IB'bulky/IIIB, 95 por ciento carcinomas escamosos) antes de la quimioterapia y a los 30 días del tratamiento. Trece pacientes recibieron un esquema de drogas basado en cisplatino y como la respuesta a esta terapia no fue buena, a las otras 11 pacientes se les administró vinorelbine e ifosfamida. Luego de la quimioterapia todas las pacientes recibieron radioterapia. La expresión de los marcadores moleculares en las biopsias pre- y post quimioterapia se estudió por inmunohistoquímica y la apoptosis fue evaluada por la técnica del TUNEL mejorada recientemente. Para comparar los cambios en la expresión de los marcadores moleculares y para correlacionarlos con la evaluación clínica (media de seguimiento: 31 meses para las pacientes que recibieron cisplatino y 19 para las que recibieron vinorelbine e ifosfamida) se realizaron análisis estadísticos. Resultados y conclusiones: La quimioterapia de inducción no aumentó la sobrevida de las pacientes, el 50 por ciento tuvo enfermedad progresiva (EP) o falleció (F). La expresión de p21waf1/sdII/cip1, hMLF1, hMSH2, y Bcl-2 no mostró cambios significativos después de la quimioterapia y no correlacionó con la evaluación clínica. La expresión de p53 no se modificó luego de la quimioterapia, las pacientes con tumores p53 positivos mostraron una tendencia a tener una sobrevida menor. Las pacientes con EP o que fallecieron mostraron niveles altos de PCNA, a diferencia de aquellas que estuvieron libres de enfermedad (LE) o con enfermedad estable (EE) (50 por ciento versus 17 por ciento, respectivamente, p<0.004). La sobrevida de las pacientes con bajos índices de TUNEL (igual o menor al valor medio entre las biopsias pre y post-quimioterapia de 1.5) fue significativamente más corta que las pacientes que presentaron índices de TUNEL altos (p<0.009). Nuestros resultados muestran que la quimioterapia de inducción (los dos tratamientos aplicados en este estudio) no mejoró la sobrevida de pacientes con cáncer de cervix...

Cyclin D1 Expression in Primary Breast Carcinoma: Correlation with Estrogen Receptor Status and Other Clinicopathologic Parameters

PURPOSE: Cyclins are groups of proteins that play a role as a major regulator of the G1 restriction point promoting inactivation of the retinoblastoma protein. The cyclin D1 gene, CCND1, is amplified in approximately 20% of breast carcinomas and the protein is reportedly overexpressed in 60~80% of all cases. Cyclin D1 overexpression was strongly correlated to estrogen receptor positivity and better histologic grade in breast cancer. The aim of this study was to correlate cyclin D1 overexpression using a well characterized antibody with estrogen receptor status and other clincopathologic parameters. METHODS: From March 1989 to December 1994, 85 patients with primary breast carcinoma were the subject in this study. We analyzed cyclin D1 expression by immnohistochemical staining using cyclin D1 antibody, cells were considered positive according to distinct nuclear staining. The correlation between cyclin D1 expression was compared with important clinicopathologic parameters (tumor size, axillary lymph node status, p53 expression, c-erbB2 expression, histologic grade, estrogen receptor status). RESULTS: Cyclin D1 expression was detected in 37 cases (43.5%). Cyclin D1 expression was high in patients with tumors that expressed estrogen receptor (58.5% vs 26.5%, P=0.019). Cyclin D1 was mainly overexpressed in the histologic grade I and II (75.0%), as compared with 65.2% in cyclin D1 negative tumor, however there was no statistical significance (P=0.067). There were no significant correlation with tumor size, axillary lymph node status, p53 expression, or c-erbB2 expression (P>0.05). CONCLUSION: Cyclin D1 expression in estrogen receptor (ER) positive patients was significantly higher than that seen in ER negative patients. There was a negative correlation between cyclin D1 and tumor histologic grade, however it was not statistically significant. Tumor size, axillary lymph node status, p53 expression, and c-erbB2 expression were not correlated with cyclin D1.

Expression of beta-Catenin, c-Myc, and Cyclin D1 in Pulmonary Adenocarcinomas

BACKGROUND: beta-Catenin has dual functions: adhesive molucule and transcriptional activator. Subcellular accumulation of beta-catenin and subsequent formation of beta-catenin- Tcf/Lef-1 complexes, as well as c-myc and cyclin D1 genes which were recently defined as target genes of beta-catenin- Tcf/Lef-1, has been shown to be important in the development of colorectal and breast carcinomas. The author investigated the rate of subcellular accumulation of beta-catenin and overexpression of c-myc and cyclin D1, and also investigated the association between them in the pulmonary adenocarcinomas. METHODS: Fifty-one surgically resected primary adenocarcinomas of the lung, including 11 bronchioloalveolar carcinomas, were investigated by immunohistochemical analysis with monoclonal antibodies specific for beta-catenin, c-myc and cyclin D1. Clinico-pathological information were collected from the patient charts and surgical pathology reports. RESULTS: Accumulation of beta-catenin in the nucleus and/or cytoplasm and overexpression of c-myc and cyclin D1 were observed to be 20%, 37%, 16%, respectively. Ten cases showing accumulated patterns of beta-catenin revealed alternative overexpressions of c-myc (7 cases) and cyclin D1 (3 cases). In nonmucinous tumors, 9 cases showing overexpression of c-myc or cyclin D1 revealed accumulations of beta-catenin. The accumulation of beta-catenin was not statistically related to clinico-pathological parameters. The association between c-myc overexpression and histological subtype of tumors was observed. CONCLUSIONS:It is suggested that the accumulation of beta-catenin is closely associated with tumorigenesis in a minor subset (20%) of peripheral adenocarcinomas of the lung. It is also suggested that transactivation of beta-catenin may closely be associated with the overexpression of c-myc or cyclin D1 in the nonmucinous adenocarcinoma.

Cyclin D1 Gene Amplification in Oral Squamous Cell Carcinoma using Differential Polymerase Chain Reaction

Neoplastic growth is characterized by alterations of oncogenes and antioncogenes. The interaction between activated oncogenes and functional deletion of antioncogene appears to be the driving force directing normal cells to uncontrolled growth resulting in tumor. In addition to those genes mentioned, other genes controlling the entry of cells into the cell cycle have recently been implicated in cancer development. The overexpression of the cyclin D1 gene, which has been mapped to 11q13, either by gene rearrangement or amplification has been noted in various malignant tumors. The product of the cyclin D1 gene forms a complex with cyclin-dependent protein kinases(CDK4) that governs a key transition in the cell cycle. The relationships between the overexpression of cyclin D1 assessed by immunihistochemistry and the amplification of the cyclin D1 gene by differential polymerase chain reaction(DPCR) using primers for dopamin D2 receptor gene in 13 cases of squamous cell carcinomas of the oral cavity have been studied. The semiquantitative assay of cyclin D1 amplification has been made by cyclin D1/dopamin D2 receptor(CD/DR) ratio. The results were as follows; 1. In the normal tissue and the tumor, the CD/DR ratios were 0.82 and 1.36 respectively. This implicates 1.65-fold amplification of cyclin D1 gene in tumor compared to that in normal tissue. 2. The tumor tissue which showed overexpression of cyclin D1 by immunohistochemistry revealed 2-fold amplification of cyclin D1 compared to the normal tissue. 3. The tumor tissue which showed mild expression of cyclin D1 by immunihistochemistry revealed 1.7-fold amplification of cyclin D compared to the normal tissue. 4. The cyclin D1 was overexpressed in the tumor tissue at the rate of 38%. Above results suggest that cyclin D1 has close correlation with the development of carcinoma in the oral cavity. But further studies were needed to elucidate the carcinogeneic mechanisms by comparative studies among cyclin D1, pRb and p53.

Prognostic Significance of Cyclin D1 Overexpression in Non-Small Cell Lung Cancer / 결핵

BACKGROUND: The cyclin D1 gene is one of the most frequently amplified chromosomal regions(11ql3) in human carcinomas. In laryngeal and head and neck carcinomas, its overexpression has been shown to be associated with advanced local invasion and presence of lymph node metastases. Cyclin D1 may therefore play a key role in cell growth regulation and tumorigenesis. Lung cancer is a worldwide problem and in many contries it is the most lethal malignancy. As relapse is frequent after resection of early stage non-small cell lung cancer, there is an urgent need to define prognostic factors. PURPOSE: This study was undertaken to evaluate the prognostic value of the cyclin D1, that is one the G1 cyclins which control cell cycle progression by allowing G1 to S phase transition, on the patients in radically resected non-small cell lung cancer. METHOD: Total Si cases of formalin-fixed paraffin-embedded blocks from resected primary non-small cell 11mg cancer from January 1., 1983 to July 31, 1995 at Hanyang University Hospital were available for both clinical follow-up and immunohistochemical staining using monoclonal antibodies for cyclin D1. RESULTS: The histologic classification of the tumor was based on WHO criteria, and the specimens included 45 squamous cell carcinomas, 25 adenocarcinomas and 11 large cell carcinomas. Cyclin D1 overexpression was noted in 26 cases of 81 cases tested (30.9%). Cyclin D1 expression was not significantly associated with cell types of the tumor, pathological staging and the size of the tumor. But cyclin D1 overexpression was significantly correlated with positive lymph node metastasis(p=0.035). The mean survival duration was 22.76+/-3.50 months in cyclin D1 positive group and 45.38 +/- 5.64 months in cyclin D1 negative group. There was a nearly significant difference in overall survival Between cyclin D1 positive and negative groups(p=0.0515) in radically resected non-small cell lung cancer. CONCLUISON: Based on this study, cyclin D1 overexpressiom appears at important poor prognostic indicator in non-small cell lung cancer and may have diagnostic and prognostic importance in the treatment of resectable non-small cell lung cancer.

Amplification and Overexpression of Cyclin D1 in Head and Neck Squamous Cell Carcinomas / 대한이비인후과학회지

BACKGROUND: Overexpression of the cell cycle control gene, cyclin D1 may, at least in some tumor types, provide a prognostic marker. Cyclin D1 is expressed during the G1 phase of the cell cycle and becomes associated with its catalytic partner CDK4 or CDK6. This association may overcome the G1 arrest. OBJECTIVES: To establish the frequency of cyclin D1 protein overexpression and to evaluate its correlation with cyclin D1 gene amplification and its correlation with clinico-pathologic variables that are used in clinical practice. MATERIALS AND METHODS: The cyclin D1 gene amplification was estimated with the differential polymerase chain reaction(PCR) and cyclin D1 protein overexpression was evaluated with immunohistochemical study in 32 cases of resectable head and neck squamous cell carcinoma(SCC). The presence or absence of cyclin D1 protein overexpression was correlated with anatomical sites, T stage, nodal involvement, pathologic grade and survival rate. RESULTS: Six SCC cases(18.7%) showed the amplification of cyclin D1 gene. A highly statistical correlation between cyclin D1 gene amplification and cyclin D1 protein overexpression was noted(p0.05). But overexpression of cyclin D1 protein was associated with a poor survival of these cases(p<0.05). CONCLUSIONS: Overexpression of cyclin D1 is the independent prognostic factor for the head and neck squamous cell carcinoma. The cyclin D1 gene amplification results in the overexpression of cyclin D1 protein. But additional genetic mechanisms are involved in the protein overexpression. Therefore, cyclin D1 oncogene may be important in tumorigenesis in the head and neck squamous cell carcinoma.