Objective To investigate proliferating cell nuclear antigen (PCNA) gene expression in retinal pigment epithelium (RPE) cells and inhibition of antisense oligonucleotides(AS-OND) encoding PCNA mRNA to gene expression and proliferation of RPE cells, so as to search for new genetic therapy way for pro1iferative vitreoretinopathy (PVR). Methods (1) Rabbit RPE cells cultured in vitro were detected for PCNA expression by streptoavidin-biotin-enzyme complex (SABC) immunohistochemistry at several times. (2) The liposome-mediated synthetic antisense oligodeoxynucleotides (AS-ODN) and sense oligodeoxynucleotides (S-ODN) encoding PCNA were delivered to the RPE cells at different concentrations, then PCNA expresstion were detected by immunohistochemistry. (3) Exposed to different concentrations of AS-ODN and S-ODN, growth activity and suppressive rate of RPE cells were measured by methyl thiazolyl tetrazolium (MTT) methods. Results (1) PCNA were expressed in RPE cells, culmination in 48 hours of culture. (2) PCNA expression were markedly suppressed in the RPE cells treated with 0.28 and 1.12 μmol/L PCNA AS-ODN. (3) 0.28 μmol/L and 1.12 μmol/L PCNA AS-ODN significantly inhibited proliferative activity of RPE cells in a dose-dependent manner, the arrest rates of cellular growth reached 53% and 81% respectively. Conclusion AS-ODN complementary to PCNA mRNA at some concentration can sequence-specifically suppress PCNA expression in RPE cells and cellular proliferative activity, and show potential application to further experimental study for PVR genetic medication. (Chin J Ocul Fundus Dis, 2002, 18: 231-233)
PURPOSE:To investigate the relationship between the proliferative activity of refinoblastoma (RB)cell and the RB differentiation degree and the infiltration capability. METHOD:The proliferating cell nuclear antigen (PCNA)expression in RB tissues of 48 cases was analysed by using LSAB immunohistochemical method. RESULTS :The mean PCNA labelling index(LI)in differentiated RB tissues of 12 cases was markedly lower than that in non-differentiated of 36 cases(P＜0.05). The mean PCNA LI in RB tissues of the optic nerve infiltrated group(22 cases)was significantly higher than that of the optic nerve non-infiltrated group(26 cases)(P＜0.05). The results indicate that the PCNA LI is significantly related with the differentiation degree of RB and the infiltration capability. CONCLUSION :The determination of PCNA LI is of significance for evaluating the histologic characteristics and biological behavior of RB.
Purpose To observe the expression of proliferating cell nuclear antigen（PCNA）and bcl-2 of cultured human retinal pigment epithelial cells（RPE）． Methods SABC techniques were applied for immunocytochemical staining of cultured RPE with mouse anti-human PCNA monoclonal antibody and rabbit antihuman bcl-2 antibodies． Results 31.2% and 50.6% cultured cells were positive to anti-human PCNA at 24h and 48h after seeding，respectively．The positive staining was mottled in the nucleus．positive staining for bcl was seen in 76%to 90% cells as fine granules scattered within the cytoplasm. Conclusion One half of cultured RPE expressed PCNA，indicating that the cells were in phase S of the cell cycle．Positive staining for bcl-2 appeared in much more RPE cells．These biological markers may be associated with the growth activity of cultured RPE． (Chin J Ocul Fundus Dis,1998,14:26-28)
Objective To examine the expression of proliferating cell nuclear antigen (PCNA) of retinal pigment epithelial (RPE) cells, thus assessing the role of mechanism of contact inhibition playing in the process of experimental retinal detachment and reattachemnt.Methods Retinal detachment was produced in 72 cats by subretinal injection of 0.25% solution of healon through a micropipette three weeks after extracapsular lens extraction and vitrectomy. Some of the detached retinae were reattached 24 hours later. At different time, the cats were killed and eye globes were fixed and embeded in paraffin. Histologic sections were processed for immunohistochemistry examination using an antibody to detect PCNA protein. Labeled RPE cells were identified, and the proliferation was quantified in detached and un-detached retinae of detachment group, and also in reattached retinae of reattachment group. The comparsion of PCNA-labeled RPE cells in different groups were analyzed by ANOVA. Results In detached regions of detachment group, PCNA-expression of RPE cells occured within 24 hours, and reached a maximum after 5-6 days, then gradually declined to barely detectable levels after 20 days. Similar tendency was found in reattached retinae, but the number of PCNA-labeled RPE cells was obviously small. Fewer PCNA-labeled RPE cells were found in regions of un-detached retinae in detachment group. The difference of these three groups was significant.Conclusion Proliferation of RPE cells is induced when they lose contact with neural retina, but inhibited after neural retina reattached to RPE cells. It suggests that the mechanism of contact inhibition plays a role in the proliferative process after retinal detachment and reattachment. (Chin J Ocul Fundus Dis，2003，19：20-23)
Objective To explore the value of expression of carcinomaassociated antigens in early diagnosis and predicting prognosis in gallbladder carcinoma. MethodsThe expression of carcinoembryonic antigen (CEA), carbohydrate antigen (CA50), Ecadherin (ECD) and proliferating cell nuclear antigen (PCNA) in 10 cases of cholecystitis, 10 cases of gallbladder adenomas and 50 cases of gallbladder carcinomas were detected by immunohistochemistry. ResultsThe positive rate of CEA, CA50 and PCNA labeling index (LI) in gallbladder carcinomas were significantly higher than that of gallbladder adenomas and cholecystitis (P＜0.05 and P＜0.01). The positive rate of ECD in gallbladder carcinomas, especially with metastasis, was significantly lower than that of gallbladder adenomas and cholecystitis (P＜0.05). The 3year survival rate was significantly lower in gallbladder carcinomas with CEA and PCNA overexpression (P＜0.05), the 3year survival rate in patients with ECD positive tumors was higher than that of those with negative tumors (P＜0.05). Conclusion The detection of CEA, CA50 and PCNA is useful for early diagnosis of malignant change in gallbladder adenomas and gallbladder carcinomas. Therefore, the CEA, PCNA and ECD might be useful for predicting prognosis of gallbladder carcinomas.
Objective To examine the relationship between ratio of proliferating cell nuclear antigen (PCNA)/apoptosis and biology of hepatocellular carcinoma. Methods Thirty five cases of hepatocellular carcinoma were studied with TUNEL and immunohistochemistry. Results Positive rate of apoptosis in grade Ⅰ, Ⅱ, Ⅲand Ⅳ was 1.37%,0.70%, 0.67% and 0.25% respectively. Positive rate of PCNA in grade Ⅰ, Ⅱ, Ⅲ and Ⅳ was 17%, 19%, 75% and 80%. Ratio of PCNA/apoptosis in grade Ⅰ,Ⅱ,Ⅲ and Ⅳ was 19.04, 75.51, 138.01 and 345.52. Conclusion Ratio of PCNA/apoptosis is correlated with histological classification.
Objective To investigate the effect of alltrans retinoic acid (atRA) on proliferative artery disease after heart transplantation. Methods Heterotopic heart transplantation model was established by Ono model with 16 inbred healthy male Wistar rats as donors and 16 SD rats as recipients. The rats were divided into chronic rejection group and atRAtreated group by complete random design, and there were 8 rats in each group. Rats in chronic rejection group were given Cyclosporine A 10 mg/(kg·d) by subcutaneous injection after operation, and those in atRAtreated group were given Cyclosporine A 10 mg/(kg·d) in the same way and atRA 10mg/(kg·d) by gavage. The transplanted hearts of rats were taken out 60 days after the transplantation. HE stain, masson stain and Van Gieson were done to analyze the rejection of transplanted hearts, the degree of vascular stenosis and myocardial fibrosis respectively.Immunohistochemistry was used to test proliferating cell nuclear antigen (PCNA). Results The area of myocardial fibrosis in chronic rejection group was obviously larger than that in atRAtreated group（63.99%±11.91% vs.34.68%±6.34%）, and there was significant difference between two groups（t=8.377，P=0.000）. The index of vascular stenosis in chronic rejection group was higher than that in atRAtreated group（62.86±17.18 vs. 40.10±8.20）. Vascular stenosis in atRAtreated group alleviated significantly, and there was significant difference between two groups（t=3.913, P=0.006）. The PCNA positive cells in chronic rejection group were obviously more than that in atRAtreated group（60.17±17.74 vs. 33.96±8.65）, and there was significant difference between two groups（t=5.387, P≤0.001）. There was a positive correlation between the PCNA positive cell ratio and the index of vascular stenosis（r=0.854, P=0.007）. Conclusion Alltrans retinoic acid can inhibit vascular disease after heart transplantation by cell proliferative pathway.
【Abstract】Objective To investigate the inhibitory effects of flavonoids quercetin on the occurrence and proliferation of experimental mammary carcinoma. Methods DMBA induced mammary carcinoma was produced in rats. Seventy-nine female Sprague-Dawly rats were divided randomly into four groups: DMBA, DMBA with TAM, DMBA with quercetin and control. Chemicals had been administered to group A, group B, group C and group D respectively for 28 weeks. Samples of breasts were collected for light microscope observation and electromicroscope observation. Their expressions of proliferating cell nuclear antigen (PCNA) and the protein product of H-ras were examined by immunohistochemical staining. Results ①Mammary carcinoma incidence of group A(76.2%) was significantly higher than that of group B(40.9%), group C(45.5%) and group D(0%),P＜0.05, and there was no significant difference between group B and group C (P＞0.05), which indicated that quercetin could inhibit the occurrence of mammary carcinoma. ②Mean mammary tumor diameter of group A (2.37cm) was significantly larger than that of group B(1.82cm) and group C(1.71cm), P＜0.05, and there was no significant difference between group B and group C (P＞0.05), which indicated that quercetin could inhibit the growth of experimental mammary carcinoma. ③Immunohistochemical staining of PCNA showed significant difference between group A and group B, group A and group C (P＜0.05), with no significant difference between group B and group C (P＞0.05), which indicated that quercetin could inhibit the proliferation rate of tumor cells. ④Significant difference between group A and group B, group A and group C (P＜0.05), and no significant difference between group B and group C (P＞0.05), were noticed with immunohistochemical staining of H-ras protein product, which indicated that quercetin could inhibit the activity of Hras protein. Conclusion Quercetin could reduce the mammary carcinoma incidence and its degree of growth, and it may be related with its inhibitory effect on the activity of Hras and the proliferation of tumor cell.
To determine the state of fibroblast during the process of development of hypertrophic scar (HS), 40 specimens of HS in different periods were collected. The expressions of prolifrating cell nuclear antigen (PCNA) and Ag-protein in nucleolar organizer regions (Ag NORs) as well as the content of total amino acids in the tissues were examined. The hypertrophic scar of 1st and 3rd month old, the expression of PCND and Ag NORs were the highest. In the 9th and 12th month old, althrough PCNA was nearly negative, but the expression of Ag NORs was low. The content of total amino acid was increased gradually as HS developed but the increase of amount of hydroxyproline was markedly slowed down in 9 month old HS. It was suggested that: (1) in the developing process of HS the proecess of overproliferation of fibroblasts was short and limitted in 1-3 months period in the process of wound lealing; (2) the synthesis of collagen was nearly stopped at 6 months, but that of other extracellular matrix such as fibronectin and proteoglycan might be continued to aggregate after 12 months.
ObjectiveTo study the expression of survivin protein in primary hepatocellular carcinoma(PHC) and its relationship to the proliferation of the tumor cells and prognosis of PHC. MethodsThe expression of survivin protein and the proliferation of tumor cells marked by proliferating cell nuclear antigen (PCNA) in 48 cases of PHC were determined by immunohistochemical method. ResultsThe survivin protein was expressed in 31 of 48 cases of PHC (64.6%). The expression of PCNA was significantly higher in hepatocellular carcinoma (HCC) with positive survivin expression than in HCC with negative survivin expression. The patients with positive survivin expression had the worse prognosis than those with negative survivin expression. ConclusionThe expression of survivin may play an important role in the proliferation of PHC cells and closely associate with the prognosis of PHC, and probably become the prognostic factor and an important target of therapy.