PURPOSE: Peroxisome proliferator-activated receptor gamma (PPARgamma) has become a potential target for the prevention and treatment of human cancers. PPARgamma ligands inhibit cell proliferation of estrogen receptoralpha(ERalpha)-positive breast cancer cells. However, it has recently been shown that ERalpha-negatively inhibits PPARgamma signaling in breast cancer cells, indicating that PPARgamma ligand may be more useful for treating ERalpha-negative breast cancer cells compared to ERalpha-positive breast cancer cells. In this study, we attempted to elucidate the role of PPARg in ERalpha-negative breast cancer cells. METHODS: The effect of PPARgamma ligand on the growth of MDA-MB-231 cells was measured by MTT assay and flow cytometric analysis. TUNEL staining and Hoechst 33342 fluorescent staining were used to observe the effects of PPARgamma ligand on cell apoptosis. The regulatory proteins of the cell cycle were measured by Western blot. RESULTS: The treatment of MDA-MB-231 human breast cancer cells with the PPARgamma ligand, trgoglitazone, was shown to induce inhibition of cell growth in a dose-dependent manner. Cell cycle analysis showed a G1 arrest in MDA-MB-231 cells exposed to troglitazone. The apoptotic effect by troglitazone demonstrated that apoptotic cells were elevated from 2.5-fold of the control level at 10 mM, to 3.1-fold at 50micrometer and to 3.5-fold at 75 mM of troglitazone. Moreover, troglitazone treatment dose-dependently caused a marked decrease in the pRb, cyclin D1, cyclin D2, cyclin D3, cdk2, Cdk4 and Cdk6 expressions and there was a significant increase in the p21 and p27 expressions. CONCLUSION: These results indicate that trgoglitazone induces cell-cycle G1 arrest and apoptosis in ERalpha-negative MDA-MB-231 breast cancer cells. Collectively, this paper shows that PPARgamma ligand is an important player as a member of the chemotherapeutic candidates for treating ERalpha-negative breast cancer.
Apoptosis* ; Blotting, Western ; Breast Neoplasms* ; Breast* ; Cell Cycle ; Cell Proliferation ; Cyclin D1 ; Cyclin D2 ; Cyclin D3 ; Estrogens ; Humans ; In Situ Nick-End Labeling ; Ligands ; Peroxisomes* ; PPAR gamma*

OBJECTIVETo clarify the role of these cyclins in human gastric cancer.

METHODS38 gastric cancer patients, 29 first degree relatives of gastric cancer patients, as well as 18 healthy subjects were included. The mRNA expression of cyclins D1, D2, D3 and E in gastric biopsies was evaluated by RT-PCR analysis using specific primers. Histomorphological features such as intestinal metaplasia, atrophy, H. pylori infection and severity of gastritis were determined by the updated Sydney System.

RESULTSSignificant mRNA overexpression was found for cyclins D2, D3 and E compared with healthy normal specimen, but cyclin D1 expression was not different between tumor and normal tissues. In addition, cyclin D2 and D3 overexpression was significantly more frequent in first degree relatives than in healthy controls (P < 0.05). Among the various pathological findings, the overexpression of cyclins D2 and E was associated with intestinal metaplasia, and the overexpression of cyclin D3 was associated with intestinal metaplasia as well as atrophy. The overexpression of cyclins D2 and D3 was significantly correlated with H. pylori infection. No correlation was observed between the overexpression of cyclin D1 and any pathological variables.

CONCLUSIONThe overexpression of cyclins D2, D3 and E is a frequent event in patients with gastric cancer and their first degree relatives and may be an early event in gastric carcinogenesis.

Adult ; Aged ; Aged, 80 and over ; Cyclin D1 ; genetics ; Cyclin D2 ; Cyclin D3 ; Cyclin E ; genetics ; Cyclins ; genetics ; Family Health ; Gastritis ; genetics ; Gene Expression Regulation, Neoplastic ; Helicobacter Infections ; genetics ; microbiology ; Helicobacter pylori ; growth & development ; Humans ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Stomach ; metabolism ; microbiology ; pathology ; Stomach Neoplasms ; genetics ; pathology

Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; 0.1 microgram/ml; 1 microgram/ml; 10 microgram/ml; 100 microgram/ml; 1000 microgram/ml. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of 1 microgram/ml - 1000 microgram/ml, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of 1 microgram/ml - 1000 microgram/ml and at 10 microgram/ml - 1000 microgram/ml respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of 10 microgram/ml - 1000 microgram/ml. Treatment with 100 microgram/ml nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin D1 and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin D1 and CDK 4 in human gingival fibroblasts.
Blotting, Western ; Cell Cycle Proteins* ; Cell Cycle* ; Cell Proliferation ; Cell Survival ; Connective Tissue ; Cyclin D ; Cyclin D1 ; DNA ; Fibroblasts* ; Humans* ; Nicotine* ; Propidium ; Tobacco ; Wound Healing

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.
Carcinoma, Squamous Cell* ; Cell Cycle ; Cyclin D ; Cyclin D1* ; Cyclins* ; Gene Rearrangement ; Genes, bcl-1* ; Genes, Tumor Suppressor ; Immunohistochemistry ; Mouth ; Oncogenes ; Polymerase Chain Reaction*

BACKGROUND: There is experimental evidence that overxpression of cyclin D1 accelerates the entry of cells into S-phase, but that p16 inhibits the CDK4 and CDK6 by binding in competition with cyclin D1 . Previous attempts to correlate cyclin D1 amplification with prognoses have frequently drawn associations with adverse outcome or a more aggressive phenotype. Recently, overexpression of cyclin D1 has been associated with improved relapse-free survival and overall survival rates. To elucidate whether the expression of cyclin D1 and p16 protein might be of clinical value as prognostic factors, the immunoreactivity of cyclin D1 and p16 protein were compared by the chi-square test with histopathologic findings and such known prognostic factors as estrogen receptor, progesteron receptor, c-erbB-2, p53 and Ki-67. METHODS: The expression of cyclin D1 and p16 protein was analysed using immunohistochemical methods in formalin-fixed and paraffin-embedded tissue samples of 340 invasive breast carcinomas accumulated between 1990 to 1997 at Yeungnam University Hospital. Disease-free survival and overall survival were compared to cyclin D1 and p16 status by Kaplan-Meier method. RESULTS: The nuclear immunoreactivity of cyclin D1 and P16 protein was detected in 75.6% (257/340) and 70.5% (208/295) cases, respectively. Cyclin D1 was found to have a strong correlation with lower histologic grade, lower nuclear grade, lower mitotic index, lower SBR and MSBR grade(p<0.05). Cyclin D1 was more common in non-ductal carcinomas than ductal carcinomas, though this did not reach statistical significance. Cyclin D1 was also correlated with positive estrogen receptor, negative c-erbB-2 and positive p16 protein. P16 protein expression was found to have a correlation with positive estrogen receptor and progesterone receptor. The expression of cyclin D1 and p16 protein was not significantly correlated with overall survival and disease free survival. CONCLUSIONS: These results showed that expression of cyclin D1 and p16 protein could not be used as a prognostic indicator in primary breast carcinoma.
Breast Neoplasms* ; Breast* ; Carcinoma, Ductal ; Cyclin D ; Cyclin D1* ; Cyclins* ; Disease-Free Survival ; Estrogens ; Mitotic Index ; Phenotype ; Prognosis ; Receptors, Progesterone ; Survival Rate

A low serum level of vitamin D has been associated with an increased incidence of gastrointestinal tract cancers. However, the effects of vitamin D3 have not been investigated in gastric cancer and cholangiocarcinoma. In the present study, we found that vitamin D3 treatment significantly suppressed the viability of gastric cancer and cholangiocarcinoma cells. Moreover, vitamin D3 had a synergistic effect with other anti-cancer drugs, such as paclitaxel, adriamycin, and vinblastine, for suppressing cell viability. To determine the underlying mechanism involved in the regulation of viability by vitamin D3, we examined the effects of vitamin D3 on expression of hedgehog signaling target genes, which has been associated with gastric cancer and cholangiocarcinoma. Vitamin D3 treatment decreased the level of mRNA expression of patched1, Gli1, cyclin D1, and Bcl2, suggesting the possibility that vitamin D3 may act through regulation of hedgehog signaling. From the above results, we conclude that vitamin D3 regulates cell viability in gastric cancer and cholangiocarcinoma.
Cell Survival ; Cholangiocarcinoma ; Cholecalciferol ; Cyclin D1 ; Doxorubicin ; Gastrointestinal Neoplasms ; Hedgehogs ; Incidence ; Paclitaxel ; RNA, Messenger ; Stomach Neoplasms ; Vinblastine ; Vitamin D ; Vitamins

BACKGROUND: The purpose of this study is to assess the roles of chromogranin A, cathepsin D, cyclin D1 and p53 protein expression in colorectal tumorigenesis. METHODS: 83 colorectal cancer and 12 villotubular adenoma tissue specimens were investigated by immunohistochemical staining for chromogranin A, cathepsin D, cyclin D1 and p53 protein. Clinicopathologic values (tumor size, histologic grade, Astler-Coller stage and lymph node metastasis) were compared with the incidence of chromogranin A, cathepsin D, cyclin D1 and p53 protein expression in colorectal adenocarcinomas. RESULTS: Statistically significant correlation was noted between the expression of chromogranin A and histologic grade (p<0.05). The incidence of positive cathepsin D expression was increased with tumor size (p<0.05), and there was a statistically significant correlation between histologic grade and cathepsin D expression (p<0.005). There were no statistically significant correlations among cyclin D1 expression and tumor size, histologic grade, stage and lymph node metastasis. Patients with lymph node metastasis had a high incidence of positive p53 protein expression compared to those without this finding (p<0.001). CONCLUSION: It is suggested that chromogranin A, cathepsin D, and p53 protein are useful variables for the prognostic assessment of colorectal adenocarcinoma. The p53 protein seems to involve the metastatic ability of colorectal adenocarcinomas. Also, the expression of cathepsin D, cyclin D1, and p53 protein may play an important role in the tumorigenesis and progression of the colorectal adenoma-carcinoma sequence.
Adenocarcinoma* ; Adenoma ; Carcinogenesis ; Cathepsin D* ; Cathepsins* ; Chromogranin A* ; Colorectal Neoplasms ; Cyclin D1* ; Cyclins* ; Humans ; Incidence ; Lymph Nodes ; Neoplasm Metastasis

PURPOSE: Biologic characteristics of the tumors could be altered after chemotherapy. Accurate assessment of tumor biology before treatment is important for selecting treatment modalities. Current study was designed to investigate whether multiple molecular markers could be accurately assayed on the fine needle aspirates from the breast carcinoma. METHODS: Immunocytochemical assays (ICA) for p53, cyclin D1, and cathepsin D were performed on cytologic samples from 76 primary breast carcinomas, 37 ductal carcinoma in situ (DCIS), and 36 benign ductal hyperplasia. ICA for 3 molecules was also performed on the histologic sections from the matching tumor blocks, and the results were compared. RESULTS: Three molecular markers were successfully detected in cytologic samples from the breast carcinomas; p53 in 71.1% (54/76), cyclin D1 in 73.7% (56/76), and cathepsin D in 44.7% (34/76). Their expression was rarely observed in benign hyperplasia; p53 in 0/36, cyclin D1 in 7/36, and cathepsin D in 4/36. Increase of expression frequency of 3 molecules was apparent through the progress of the disease. Results of ICA for each molecules were well correlated between cytologic and histologic samples; concordance was 93.4% (71/76) for p53, 81.6% (62/76) for cyclin D1, and 73.7% (56/76) for cathepsin D. When 3 molecular markers were integrated to the preoperative diagnosis, positive predictive value was 90.6% for malignant breast disease. CONCLUSION: Three molecular markers were successfully assayed on cytologic samples and well correlated with the results from the histologic sections. The results indicate that the biologic information from the fine needle aspirates can be reliably integrated to patients treatment.
Biology ; Breast Diseases ; Breast Neoplasms* ; Breast* ; Carcinoma, Intraductal, Noninfiltrating ; Cathepsin D* ; Cathepsins* ; Cyclin D1* ; Cyclins* ; Diagnosis ; Drug Therapy ; Humans ; Hyperplasia ; Needles ; Population Characteristics

BACKGROUND AND OBJECTIVES: Various host and tumor parameters, particularly the tumor size and lymph node metastasis have been studied in an attempt to evaluate and decide the optimal treatment of the patients with head and neck carcinomas. Moreover, it has been recognized that prognostic parameters can be useful for the evaluation of biological behaviors of malignancy. The p53 is a tumor suppressor gene and cyclin D1 is a cell cycle regulator, essential for G1 phase progression. Cathepsin D is a lysosomal aspartyl endopeptidase which degrades the extracellular matrix and proteoglycan. But there are still controversy in their clinical meanings in sinonasal malignant tumors. The purpose of this study is to assess the roles of p53, cyclin D1 and cathepsin D in sinonasal tumorigenesis. MATERIALS AND METHOD: 27 inverted papilloma (IPs), 5 IPs associated with malignant transformation, and 16 squamous cell carcinoma tissue specimens were investigated by immunohistochemical staining for p53, cyclin D1, and cathepsin D. Clinicopathologic values were compared with the incidence of p53, cyclin D1, cathepsin D expression in sinonasal malignant tumors. RESULTS: p53/cyclin D1 expressions were increased as tumor progressed and these expressions were statistically significant (p< .05). No significant correlations were found among p53, cyclin D1, cathepsin D and other clinicopathologic factors. CONCLUSION: These data suggest that expressions of p53, cyclin D1 and cathepsin D may play an important role in the tumorigenesis and progression of sinonasal malignant tumor sequence. Also, it is suggested that p53/cyclin D1 expressions may be useful variables for the prognostic assessment of sinonasal malignant tumors. However, it is not enough conclude so based on this result alone. Further studies, such as using molecular biological techniques, will be required to determine that p53/cyclin D1 expressions are related to the development or prognosis of sinonasal malignant tumors.
Carcinogenesis ; Carcinoma, Squamous Cell ; Cathepsin D* ; Cathepsins* ; Cell Cycle ; Cyclin D1* ; Cyclins* ; Extracellular Matrix ; G1 Phase ; Genes, Tumor Suppressor ; Head ; Humans ; Incidence ; Lymph Nodes ; Neck ; Neoplasm Metastasis ; Papilloma, Inverted ; Paranasal Sinus Neoplasms ; Prognosis ; Proteoglycans

OBJECTIVETo investigate the expression of five kinds of cyclins in hepatocellular carcinoma (HCC) and their association with degree of tumor differentiation, metastasis and infection of hepatitis B virus (HBV).

METHODSThe HCC tissue microarrays were composed of those from 273 cases of HCC tissues, 144 surrounding-tumor liver tissues and 10 normal liver tissues obtained from autopsy. The diameter of each specimens on tissue microarrays was 2.0 mm. Immunohistochemistry was used to detect the expression of cyclin A, cyclin B, cyclin D1, cyclin D3 and cyclin E on HCC tissue microarrays. The association of the expression of these cyclins with the infection rate of HBV was also analyzed.

RESULTSThree paraffin-embedded HCC tissue microarrays were successfully constructed, including 136, 143 and 148 tissue spots, respectively. The positive rates of cyclins in 273 cases of HCC were cyclin A 52.7%, cyclin B 45.4%, cyclin D1 35.9%, cyclin D3 44.3% and cyclin E 23.1%; while the figures in 144 surrounding-tumor tissues were 8.3%, 5.6%, 4.9%, 6.3% and 1.4%, respectively. In 10 normal liver tissues these cyclins exhibited negative staining, with the exception that cyclin D1 was positive in one case of normal liver tissue. The positive rate of cyclins in HCC were significant higher than those in surrounding-tumor liver tissues (P < 0.01), in HCC tissues with histological grade II and III, the cyclins expression were stronger than that in grade I (P < 0.05). The positive rates of cyclins, except cyclin A in HCC with portal vein invasion were higher than those without portal vein invasion (P < 0.01). Infection of HBV did not have significant relationship with the expression of cyclins (P > 0.05).

CONCLUSIONCyclins in different cell cycles overexpressed at varied levels in hepatocellular carcinoma, and the increased expression of cyclins may shorten the tumor cell cycle phase, accelerate cell proliferation, and have a close relationship with HCC aggressiveness.

Carcinoma, Hepatocellular ; chemistry ; Cyclin A ; analysis ; Cyclin B ; analysis ; Cyclin D1 ; analysis ; Cyclin D3 ; Cyclin E ; analysis ; Cyclins ; analysis ; Hepatitis B ; metabolism ; Humans ; Immunohistochemistry ; Liver Neoplasms ; chemistry