PURPOSE: To investigate the prognostic role of apoptosis and to evaluate the relationship between apoptosis and apoptosis-related genes, as well as cell proliferation in renal cell carcinoma (RCC). MATERIALS AND METHODS: Apoptosis was detected by using the terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) technique in 67 formalin-fixed and paraffin-embedded RCC specimens. Immunohistochemical stainings for p53 and retinoblastoma (Rb) proteins and proliferating cell nuclear antigen (PCNA) were also conducted simultaneously. RESULTS: The apoptotic index (AI) varied from 0.2% to 25.5%. The PCNA index (PI) ranged from 2.1% to 70.3%. The expression of p53 protein was found in 31 of 67 (46.3%) cases. Abnormal expression of Rb was seen in 23 of 67 (34.3%) cases. There was a statistically significant positive correlation between AI and increasingnuclear grade (p<0.001). A significant correlation was found between AI and PI (r=0.329, p<0.01). When comparing the AI with the expression of p53 and Rb proteins, there was no significant difference. In univariate survival analysis, nuclear grade, TNM stage, PI, expression of Rb and AI were significantly associated with shortened survival. However, TNM stage was the only independent prognostic factors by multivariate analysis. CONCLUSION: The present findings indicate that apoptosis in RCC is closely associated with cell proliferation, but not with the expression of p53 and Rb proteins. In multivariate analysis, the AI does not carry an independent prognostic significance.
Apoptosis* ; Carcinoma, Renal Cell* ; Cell Proliferation* ; DNA Nucleotidylexotransferase ; Genes, vif ; Multivariate Analysis ; Proliferating Cell Nuclear Antigen ; Retinoblastoma ; Retinoblastoma Protein

To characterize cellular responses during hepatic regeneration, we examined 13 explant livers and 5 liver allografts by immunohistochemistry for cytokeratin 7, HepPar1, CD68, alpha-smooth muscle actin (alpha-SMA) and proliferating cell nuclear antigen as well as reticulin and Masson-trichrome staining. Within a week after liver damage, elongated CD68-positive cells were detected along the border of necrotic area. The number of alpha-SMA-positive cells was slightly increased along the sinusoids. Ductular proliferation or fibrosis was negligible. After one or two weeks, the size and number of CD68-positive cells were markedly increased. alpha-SMA-positive cells increased in number within lobules and portal tracts. Ductular proliferation occurred predominantly at the limiting plate or along the border of necrotic areas. After one month, necrotic parenchyma was replaced by many ductules, CD68-positive cells, alpha-SMA-positive cells. Nodules of regenerating hepatocytes and irregular fibrosis were diffusely present. Other nonparenchymal cells were not significantly changed. These observations indicate that chronological interaction between nonparenchymal and parenchymal cells occur during the course of human hepatic regeneration and suggest extensive porto-periportal fibrosis more than a few months after the onset of fulminant hepatitis is a major indicator of chronic functional impairment necessitating liver transplantation.
Actins/analysis ; Antigens, CD/analysis ; Antigens, Differentiation, Myelomonocytic/analysis ; Human ; Immunohistochemistry ; Keratin/analysis ; Liver/*cytology ; *Liver Regeneration ; Proliferating Cell Nuclear Antigen/analysis

To investigate the effects of normal human bone m arrow fibroblastoid stromal cell line (HFCL) on the proliferation of acute myeloid leukemia cell line HL-60 and expression of vascular endothelial growth factor (VEGF), establishing coculture system of leukemia cell line HL-60 and HFCL, growth data was obtained by cell counting. Mitotic index (MI) was observed under Wright-Giemsa staining. Flow cytometry and Western blot were used as assays for cell cycle and expression of proliferating cell nuclear antigen (PCNA) separately. VE GF levels were evaluated by using commercial ELISA kits. The results showed that compared with HL-60 cells without HFCL cells, the proliferation of HL-60 cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited. The MI of HL-60 cells without HFCL cells was highest followed by HL-60 cells separated by transwell and HL-60 cells in direct contact with HFCL cells. The expression of PCNA in HL-60 cells with HFCL cells were lower than HL-60 cells without HFCL cells. Meanwhile, the percentage of HL-60 cells in G1 phase cocultured with HFCL cells was higher than that without HFCL cells while the percentage of Sphase cells was lower. The levels of VEGF in HL-60 cells with HFCL cells were lower than that in HL-60 cells alone. In conclusion, the normal bone marrow fibroblastoid stromal cells inhibited the proliferation of HL-60 cells as well as the expression of VEGF.
Cell Cycle ; Cell Division ; Cell Line ; Coculture Techniques ; Fibroblasts ; physiology ; HL-60 Cells ; cytology ; Humans ; Proliferating Cell Nuclear Antigen ; analysis ; Stromal Cells ; physiology ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVETo evaluate the down stream involvement of the bile duct in hepatolithiasis.

METHODSMechanical damage to bile duct epithelia and long standing cholangitis as result of hepatolithiasis play an important role in the carcinogenesis of bile duct epithelia and stricture of the intra- and extra-hepatic bile duct. Macromorphological and microscopic changes in bile duct mucosa of 100 consecutive patients with hepatolithiasis were investigated using intra- or post-operative cholangioscopy. Biopsy specimens of lesions obtained during cholangioscopy were studied with immunohistochemical staining and flow cytometry to determine proliferative activity and DNA content. Five cases of well-proven cholangiocarcinoma were simultaneously studied as controls.

RESULTSOf the 100 patients, those with chronic cholangitis accounted for 86% (86/100), proliferative lesions 11% (11/100), adenomatous polyps 1% (1/100), and adenocarcinoma 2% (2/100). The obvious mucosal lesion associated with hepatolithiasis was located down-stream of the bile duct, predominantly in the hilar region, e.g. orifices of the right/left hepatic duct and common hepatic duct (73% mucosa lesions in the hilar region). The intensity of cancer embryonic antigen stain and the proliferative cell nuclear antigen index increased with the development of bile duct lesions. Aneuploid DNA presented mainly in the high degree malignant adenocarcinomas (> 80% of cases).

CONCLUSIONSThe obvious mucosal lesions associated with hepatolithiasis were located down-stream of the bile duct, predominantly in the hilar region (73% of mucosal lesions). The proliferative activity of examined bile duct mucosa lesions increased with the development of pathological deterioration, which may contribute to the development of hilar bile duct stricture and hilar cholangiocarcinoma.

Adult ; Aged ; Bile Ducts ; pathology ; Carcinoembryonic Antigen ; analysis ; Cholangiocarcinoma ; etiology ; Humans ; Lithiasis ; complications ; pathology ; Liver Diseases ; complications ; pathology ; Middle Aged ; Proliferating Cell Nuclear Antigen ; analysis

OBJECTIVETo investigate the expression of cluster of differentiation 44 variant 6 (CD(44V6)) and proliferating cell nuclear antigen (PCNA) in ocular squamous cell carcinomas.

METHODSStreptavidin-biotin complex (SABC) immunohistochemistry was used to explore the expression of CD(44V6) and PCNA in 35 cases of ocular squamous cell carcinomas, 20 cases of papillomas, and 11 cases of normal eyelid tissue.

RESULTSThe CD(44V6) positive rate was 62.9% (22/35) in ocular squamous cell carcinomas, 15.0% (3/20) in papillomas, but not detectable in the 11 cases of normal eyelid tissue. The positive expression rates of CD(44V6) in ocular squamous cell carcinomas were significantly higher than in benign tumors (chi(2) = 11.57, P < 0.01) or control tissue (P = 0.001), and the positive expression rates of CD(44V6) in metastasis were significantly higher than without metastasis (P = 0.049). PCNA labeling indexes (PI) in tumors with CD(44V6) expression were significantly higher than those without (t = 20.21, P < 0.01).

CONCLUSIONSOverexpression of CD(44V6) is correlated with the progress and metastasis of ocular squamous cell carcinomas. CD(44V6) protein positive staining is associated with high PI. CD(44V6) and PCNA are useful for evaluating prognosis.

Carcinoma, Squamous Cell ; chemistry ; pathology ; Eye Neoplasms ; chemistry ; pathology ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Lymphatic Metastasis ; Proliferating Cell Nuclear Antigen ; analysis ; Skin ; chemistry

The epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein whose expression is a possible cause of increased tumor cell proliferation and has recently been proposed as a prognostic parameter in some tumors. Expression of EGFR was studied immunohistochemically in 62 cases of human renal cell carcinomas to evaluate their possible prognostic roles. We also examined the correlation between EGFR expression and cell proliferation by immunohistochemical staining for proliferating cell nuclear antigen (PCNA). Fifty-six cases (90.3%) expressed EGFR, with staining largely confined to the cell membrane and cytoplasm. Staining intensity of EGFR was directly correlated with nuclear grade (p=0.000) and TNM stage (p=0.015). PCNA index was significantly higher in EGFR-positive tumors than in EGFR- negative tumors. There was a statistically significant positive correlation between PCNA index and increasing staining intensity of EGFR (p=0.000). In univariate survival analysis, EGFR expression was significantly associated with shortened survival. However, EGFR expression was not an independent prognostic factor by multivariate analysis. These findings suggest that EGFR expression may be an important cause of tumor cell proliferation in renal cell carcinoma and further studies are needed to evaluate whether EGFR expression analysis provides independent prognostic information.
Carcinoma, Renal Cell* ; Cell Membrane ; Cell Proliferation* ; Cytoplasm ; Epidermal Growth Factor* ; Glycoproteins ; Humans ; Multivariate Analysis ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor*

To investigate the effects of normal human bone marrow fibroblastoid stromal cell lines HFCL on the proliferation of multiple myeloma cell lines RPMI8226, the co-culture system of RPMI8226 with HFCL was established, the adhesion ratio was determined by MTT colorimetric assay, growth curves were determined by trypan blue exclusion, the mitotic index (MI) of RPMI8226 cell was observed by Wright-Giemsa staining. Flow cytometry and Western blot were used to study the changes of cell cycle and expression of proliferating cell nuclear antigen (PCNA), respectively. The results showed that in co-culture the adhesion ratio of RPMI8226 after 1 hour was 29.4%; the proliferation of RPMI8226 cell line in direct contact with HFCL cell line was inhibited as compared with RPMI8226 in suspension. No obvious changes were observed in RPMI8226 cell separated by transwell inserts. The percentage of G(1) phase cells of RPMI8226 cell line in direct contact with HFCL was higher than that of RPMI8226 in suspension, and the percentage of S phase cells was lower. Moreover, the MI of RPMI8226 cell line in suspension was higher than that of RPMI8226 cell line in direct contact with HFCL cell. The expression of PCNA in RPMI8226 cell line in suspension was higher than that of RPMI8226 cell in direct contact with HFCL cell. It is concluded that the normal human bone marrow fibroblastoid stromal cell HFCL inhibits the proliferation and progression of cell cycle of multiple myeloma cell line, RPMI8226.
Bone Marrow Cells ; physiology ; Cell Adhesion ; Cell Line ; Cell Proliferation ; Coculture Techniques ; Fibroblasts ; physiology ; Humans ; Multiple Myeloma ; pathology ; Proliferating Cell Nuclear Antigen ; analysis ; Stromal Cells ; physiology

OBJECTIVETo investigate expression and significance of hTERT, telomerase associated-regulation protein (TRAP) and proliferating cell nuclear antigen (PCNA) in ovarian epithelial tumors.

METHODS106 specimens of ovarian epithelial tumors and their clinical history were collected, including 54 cases of malignancy, 33 borderline cases and 19 benign tumor cases. Immunohistochemical staining for hTERT, TRAP and PCNA were performed. Follow-up information was obtained for 45 of 87 cases (malignancy in 54 and borderline malignancy in 33).

RESULTSThe expression of hTERT was significantly different between benign (4/19) and borderline (90.9%, 30/33) cases, benign and malignant (94.4%, 51/54) cases (P < 0.001), as was the expression of TRAP between benign (4/15) and malignant (77.8%, 28/36) cases (P < 0.001). The expression of hTERT and TRAP was not higher in stage III, IV ovarian cancer patients than in stage I and II (P > 0.05, P > 0.3). The expression of PCNA between benign (6.9 +/- 5.9)% and borderline (26.4 +/- 17.8)% cases, benign and malignant (51.8 +/- 22.1)% cases, and borderline and malignant cases were different, and were statistically significant (P < 0.01, P < 0.001, P < 0.05). 33 cases of borderline malignancy are all survive. In 54 cases of malignancy, 35 of them have metastasis (64.8%), including 5 cases of lymph nodes metastasis. 4 of them died (7.4%).

CONCLUSIONSThe expression of hTERT and TRAP is associated with the malignant degree of ovarian cancer, but does not correlate with stage. The expression of TRAP resembles hTERT, which may be a new tumor-associated gene. Telomerase activity is positively associated with PCNA.

DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Neoplasm Staging ; Ovarian Neoplasms ; enzymology ; pathology ; Proliferating Cell Nuclear Antigen ; analysis ; Telomerase ; analysis ; Transcription Factors ; analysis

OBJECTIVEGrowth, regeneration and reparation of gastric mucosal epithelium may relate to the expression of peptides. This study aimed to investigate the effect of pS2, TGF-alpha and PCNA in endotoxin-induced acute gastric mucosal injury in young rats.

METHODSEighteen-day-old Wistar rats were randomly injected intraperitoneally with lipopolysaccharide (LPS) (5 mg/kg) or normal saline (control). The gastric mucosal specimens were harvested 1.5, 3, 6, 24, 48, and 72 hrs after LPS or normal saline injection (n=8 each). The pathological changes of the gastric mucosa were observed by hematoxylin-eosin staining. The expression of pS2,TGF-alpha and PCNA was measured by immunohistochemistry SP method.

RESULTSGastric mucosal injuries were the most serious 6 hrs after LPS injection, characterized by massive erosion, bleeding and cord necrosis of the gastric mucosa paralleling with gastric longitudinal axis. PCNA expression in the gastric mucosa in the LPS group 3, 6, 24 and 48 hrs after LPS injection was significantly lower than that in the control group (P<0.01). pS2 expression in the gastric mucosa weakened 1.5 hrs after LPS injection, recovered to the control level at 3 hrs and was significantly higher than the control at 6, 24, 48 and 72 hrs of LPS injection (P<0.01). TGF-alpha expression in the gastric mucosa in the LPS group increased significantly 6, 24 and 48 hrs after LPS injection when compared with the control group (P<0.01).

CONCLUSIONSPCNA expression may be associated with the proliferation activity of the gastric mucosa in the process of gastric mucosal injury/reparation. pS2 and TGF-alpha might participate in the defense and reparation of gastric mucosal cells through mediating cell proliferation following acute gastric mucosal injury.

Animals ; Endotoxemia ; metabolism ; Female ; Gastric Mucosa ; chemistry ; pathology ; Immunohistochemistry ; Male ; Peptides ; analysis ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Wistar ; Transforming Growth Factor alpha ; analysis ; Trefoil Factor-2

OBJECTIVETo investigate the expression of PCNA and Bcl-2 in the traumatic brain area transplanted with embryonic brain tissue in rats.

METHODSThe cerebral contusion of rats was induced by dropping weight. The homogenates of embryonic brain tissue were transplanted into the traumatic brain area two weeks after injury. All rats were sacrificed 6 weeks after injury (4 weeks after transplantation), and their brains were examined histologically. The expressions of PCNA and Bcl-2 in the brains were analyzed by immunohistochemical methods.

RESULTSThe histology of brain presented the capillary and glia proliferation, especially in the transplantation group. No significant difference was found in the expression of PCNA between two groups. However, Bcl-2 was overexpressed in the transplantation group.

CONCLUSIONThe transplantation of the embryonic brain tissue enhances the expression of Bcl-2, which may play a neuroprotective role following traumatic brain injury.

Animals ; Brain Injuries ; metabolism ; surgery ; Brain Tissue Transplantation ; Female ; Fetal Tissue Transplantation ; Proliferating Cell Nuclear Antigen ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats