OBJECTIVETo study relation of the expression of hTERT mRNA and cyclin A, p53 protein, proliferation cell nuclear antigen (PCNA) in ameloblastoma (AB) and to investigate clinical biological characteristics of AB.

METHODSThe hTERT mRNA was detected by in situ hybridization, cyclin A, p53 protein and PCNA by SP method. Normal oral mucosa and odontogenic keratocyst (OKC) are comparition.

RESULTSThe positive ratio of hTERT mRNA was 1/7 cases in normal oral mucosa. The expression of cyclin A, p53 and PCNA in normal oral mucosa were similar, and the positive cells distributed in stratum basale. In OKC, the positive cells distributed in stratum basale and super-strrtum basale. And the positive ratio of hTERT, cyclin A, p53, PCNA in OKC was 14/16 cases, 4/32 cases, 11/25 cases, 5/9 cases, respectively. In AB, the positive ratio of hTERT mRNA, cyclin A, p53 protein and PCNA was 94.4% (51/54), 66.7% (40/60), 85.7% (42/49) and 78.1% (25/32), respectively. A strong correlation was found between hTERT mRNA with cyclin A, p53 protein and PCNA (r(s) = 0.914, 0.848, 0.804, P < 0.05).

CONCLUSIONSThe expression of hTETR mRNA, cyclin A, p53 and PCNA is different in different tissues and lesions, and correlates with cell differentiation and clinical biology behaviour. Telomerase activity is related to cumulation of p53 protein, related to cell proliferation and differentiation, regulated by cyclin A, and higher in S phase.

Ameloblastoma ; metabolism ; pathology ; Cyclin A ; biosynthesis ; Humans ; Jaw Neoplasms ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; biosynthesis ; RNA, Messenger ; biosynthesis ; Telomerase ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis

OBJECTIVETo construct a plasmid vector expressing the short hairpin RNA (shRNA) targeting proliferation cell nuclear antigen (PCNA), and to investigate its effect in vitro on the expression of PCNA, proliferation and apoptosis of human osteosarcoma cells.

METHODSA plasmid vector expressing the short hairpin RNA targeting at PCNA was constructed and transfected into human osteosarcoma cell line MG-63 by dosper liposomal method. PCNA mRNA and protein expressions were examined using RT-PCR and immunohistochemical staining, respectively. Inhibition of the cell proliferation was studied by MTT method and colony forming assay. DNA synthesis was analyzed by (3)H-TdR incorporation and the cell cycle was determined by flow cytometry. The apoptotic cells were stained with acridine orange.

RESULTSExpression of PCNA mRNA after the transfection was markedly inhibited by 80.51% with a PI value of 25.68% of that of the control group. PCNA shRNA inhibited MG-63 cell growth detected by using MTT method, with an inhibition rate of 61.78% at 48 h. DNA synthesis rate also decreased in the (3)H-TdR incorporation test. Flow cytometry analysis showed an increase of the percentage of G(0)/G(1) phase cells, along with a decrease of cell population in the S phase. The apoptosis rate of cells transfected with the plasmid vector was 16.54%.

CONCLUSIONSPCNA shRNA significantly suppresses the expression of PCNA at both mRNA and protein levels, corresponding to an inhibition of the proliferation of MG-63 cell and an increase of the cellular apoptosis.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; biosynthesis ; genetics ; Transfection

OBJECTIVETo study the significance of beta -catenin, p53 and PCNA in PJS harmatoma.

METHODParaffin sections of 29 hamartomas of 18 patient with Peutz-Jeghers syndrome, 19 patients with colorectal mucosa and 10 persons with normal mucosa were examined using LSAB. Positive cells were detected under light microscope.

RESULTSAll 10 persons with normal mucosa showed p53 negative and beta - catenin positive. The beta - catenin protein was predominantly localized to the plasma membrane. PCNA index (PI) in normal mucosa was 10.56 +/- 7.51. The PI of hamartoma and cancer was 44.57 +/- 21.15 and 32.96 +/- 18.88. The PI of the two groups was significantly higher than that of the normal group. In hamartoma, the positive cells were mainly located in the low 1/3 part of the mucous glands. The positive rate of p53 was 24.1% (7/29) in hamartomatouspolyps and 57.9% (11/19) in colorectal carcinoma (chi(2) = 5.581, P < 0.05). Abnormal expression rate of beta- catenin in colorectal carcinoma was 73.7% (14/19) and 41.3% (12/29) in hamartomatous polyps. Some epithelial cells showed nuclear localization of beta- catenin and concentration of cytoplasm. (chi(2) = 4.825, P < 0.05).

CONCLUSIONSThe proliferation activity of hamartomatous polyps increased significantly by multifactory interaction. p53 and beta- catenin play a role in the early stage of hamartoma- adenoma-carcinoma sequence but have a different mechanism in inducing colorectal cancer.

Cytoskeletal Proteins ; biosynthesis ; genetics ; Gene Expression ; Hamartoma ; genetics ; metabolism ; Humans ; Peutz-Jeghers Syndrome ; genetics ; metabolism ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Trans-Activators ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; beta Catenin

Carcinoma, Hepatocellular ; microbiology ; pathology ; Cell Line, Tumor ; Cyclin D1 ; biosynthesis ; genetics ; Helicobacter pylori ; physiology ; Humans ; Liver Neoplasms ; microbiology ; pathology ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVE: To determine the expressions of survivin and proliferating cell nuclear antigen (PCNA)in non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance. METHODS: Immunohistochemical SP method was used to detect the expressions of survivin and PCNA in 43 patients with NSCLC and 15 normal epithelial tissues of the lung. PCNA labeling proliferative index was assessed. Forty-three patients with NSCLC were followed up for more than 5 years. RESULTS: The positive expression of survivin in NSCLC (79.1%) was significantly higher than that in normal epithelial tissues of the lung (P < 0.01). The survivin expression in Stage I + II was lower than in Stage III (P < 0.05). The overall survival time was significantly shorter in patients with high survivin expression than that in patients with absent or low survivin expression. The survivin expression was not related to sex, age, tumor size and site, histological type, grade, and lymphoid node metastasis (P > 0.05). The mean proliferative index of PCNA in NSCLC was much higher than that in normal epithelial tissues of the lung (P < 0.01). A positive correlation was present between the proliferative index and the tumor size, lymph node metastase, and clinical stage (P <0.01), while a negative correlation between the proliferative index and survival time (P <0.01). There was no correlation between proliferative index and age, sex, site, histological type and grade. The proliferative index was larger in patients with moderate or strong positive survivin expression than that in patients with negative or weak survivin expression (P < 0.05). CONCLUSION Over expression of survivin and PCNA is closely correlated to the progression and prognosis of patients with NSCLC, which is helpful to evaluate the progression of cancer and to predict the prognosis of NSCLC. The up-regulation of survivin expression and its close relationship with the cell proliferation in NSCLC suggest that survivin may play an important role in the carcinogenesis and development of lung cancer.
Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; metabolism ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; genetics ; Neoplasm Staging ; Prognosis ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Survivin

To investigate the role of progesterone receptor (PR) expression in malignant melanoma (MM), PR and proliferative cell nuclear antigen (PCNA) expression were immunohistochemistrically evaluated in a series of 35 specimens of MM, and the correlation between the immunohistochemistrical findings and clinicopathological data was also analyzed. PR expression was detected in 25.7% (9/35) of the patients with MM. No PR expression was observed in nevi. PR expression was inversely correlated with PCNA expression (r=-0.353, P=0.026). PR expression was slightly increased in females, subjects aged under 55 y, those with ulceration, non-acral subtype and diagnosis delay longer than 1 y, but the difference was not statistically significant. Selective expression of progesterone receptor in malignant melanoma might be correlated with inhibited tumor growth.
Gene Expression Regulation, Neoplastic ; Immunohistochemistry ; Melanoma/*metabolism ; Models, Biological ; Prognosis ; Proliferating Cell Nuclear Antigen/*metabolism ; Receptors, Progesterone/*biosynthesis ; Receptors, Progesterone/genetics ; Skin/metabolism ; Skin Neoplasms/metabolism

OBJECTIVETo dynamically observe the effect of Shenshuai Mixture (SM) on repair of kidney in acute renal failure (ARF) rats.

METHODSMale SD rats were divided into 4 groups randomly, the normal group, the SM group, the verapamil group and the model control group. Except those in the normal group were treated with normal saline without modeling, all remaining rats, after being made into ARF model by intra-muscular injection of glycerin, were treated with SM, verapamil and normal saline respectively via gastrogavage. Renal function, renal pathology, mRNA expression of epidermal growth factor (EGF) and protein expression of proliferating cell nuclear antigen (PCNA) were detected once every day from the 1st day to the 5th day. Results (1) BUN and Scr levels increased markedly 24 hrs after modeling, but the Scr level in the two treated groups was significantly lower than that in the model group (P < 0.05). Compared with that in the model group and the verapamil group, renal function was better in the SM group on the 3rd day (P < 0.01), and approach to normal level on the 5th day. (2) Renal pathological changes alleviated in every phase of ARF in the SM group than that in the model group, especially part of tubule regeneration could be seen on the 3rd day (metaphase), and renal structure was rehabilitated on the 5th day (convalescence), prior to those in the model group. (3) At the 3rd day expression of EGF mRNA and PCNA in tubule epithelial cell (TEC) increased remarkably in the SM group, higher than those in the model and verapamil group (P < 0.05).

CONCLUSIONSM could promote renal tissue regeneration and rehabilitation, and shorten the course of ARF through up-regulating mRNA expression of EGF in TEC.

Acute Kidney Injury ; drug therapy ; physiopathology ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Epidermal Growth Factor ; biosynthesis ; genetics ; Kidney ; physiopathology ; Male ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.
Animals ; Caspase 8 ; biosynthesis ; genetics ; Cell Line ; Cyclin D1 ; biosynthesis ; genetics ; G1 Phase ; physiology ; Histones ; genetics ; metabolism ; Ki-67 Antigen ; biosynthesis ; genetics ; Male ; Mice ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Resting Phase, Cell Cycle ; physiology ; Spermatogonia ; cytology ; metabolism ; bcl-2-Associated X Protein ; biosynthesis ; genetics

OBJECTIVETo determine and compare the difference of cell apoptosis and proliferation in the transition and peripheral zones in the human prostate.

METHODSSeventeen normal prostate glands from organ donors were sampled from normal men according to McNeal/s zonal anatomy, and 20 hyperplastic transition zones obtained from prostatectomy specimens of BPH patients. Cell proliferation and Bcl-2 expression were assessed by immunostaining using PCNA and anti-Bcl-2 antibodies, while apoptotic bodies were specifically stained using TUNEL. Bcl-2 mRNA expression was detected by RT-PCR.

RESULTSIn the normal epithelium, the rates of cell proliferation and apoptosis were markedly decreased in the transition zone as compared with the peripheral zone. The proliferation index was significantly increased in the hyperplastic transition zone in BPH, while the apoptosis index significantly decreased in comparison with the normal prostate. Bcl-2 was significantly greater in the normal transition epithelium than in the peripheral zone, and over-expressed in the hyperplastic transition zone. There was a significant negative correlation between the Bcl-2 expression and the apoptosis of the epithelial cells in the hyperplastic transition zone (r(s) = -0.867, P < 0.01).

CONCLUSIONThe hyperplastic transition zone may result from both an increase of cell proliferation and a failure of cell apoptosis. Increased expression of Bcl-2 may participate in the BPH process by blocking cell apoptosis.

Adult ; Apoptosis ; Case-Control Studies ; Cell Proliferation ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Prostate ; cytology ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics

OBJECTIVETo study the effect of platelet-derived growth factor (PDGF) and proliferating cell nuclear antigen (PCNA) antisense oligodeoxynucleotides (AODN) together on inhibiting the proliferation of the stenosis of transplanted vascular.

METHODSThe left and right external iliac arteries (length 1.0 cm) of rabbits were transplanted reciprocally. The transplanted vascular were respectively soaked in liposomes, PDGF-AODN, PCNA-AODN and PDGF-AODN adding PCNA-AODN solution about 20 minute, the vascular anastomotic were sutured by 8/0 suture of soaked in AODN solution. Four weeks later, the specimens were harvested for microscopy. The pathological morphology of transplanted vascular were observed under microscope (HE). The intimal thickness and area, stenosis ratio(%) of transplanted vascular were calculate and analysed statistically among group by computer system. The number of positive cells of PDGF's mRNA in transplanted vascular wall were counted with in situ hybridization histo-cytochemistry and the number of positive cells of PCNA's protein in transplanted vascular wall were counted by S-P immunochemistry.

RESULTSThe intimal thickness and area, stenosis ratio of transplanted vascular, the number of PDGF and PCNA positive cell in PDGF-AODN adding PCNA-AODN group were significantly lower than those in other group (P < 0.01), and that were lower evidently than PDGF-AODN group and PCNA-AODN group.

CONCLUSIONPDGF and PCNA antisense oligodeoxynucleotides together could significantly inhibit the proliferation of vascular smooth muscle cell and stenosis of transplanted vascular.

Animals ; Graft Occlusion, Vascular ; pathology ; prevention & control ; Iliac Artery ; pathology ; transplantation ; Male ; Oligonucleotides, Antisense ; genetics ; Platelet-Derived Growth Factor ; biosynthesis ; genetics ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Rabbits ; Transfection ; Transplantation, Homologous