【Abstract】Objective To find out if apoptosis is induced after intra-radiotherapy and its effects on pericarcinomal tissue. Methods From 1994 to 1998, 44 patients with unresectable liver cancer received 32P-GMS intra-radiotherapy. After 2 to 6 months the tumors in 3 cases could be resected and we used this cases as the treatment group. We use 4 patients with resectional HCC of same age, diseased region, differentiated but without anyother therapy as the control group. The TUNEL staining was used to stain the resected tissue, and the apoptosis index was counted. Results The apoptosis index of carcinoma was 29%～34%, average (31±16)% in the treatment group and that of the control group was 4%～6%, average (5±12.2)%. The apoptosis index of pericarcinomal tissue was 27%～37%, average (35±11)% in the treatment group and that of the control group was 0.3%～5%, average (4.1±3.3)%. Conclusion 32P-GMS intra-radiotherapy can enhance the apoptosis of HCC and its adjacent tissue.
【Abstract】 Objective To investigate the effect of retinoic acid (RA) on cell apoptosis and gene regulation of IGF-2in chondrocyte. Methods One 1-month-old Chinese rabbit weighted 500 g was used in this experiment. The chondrocyte from rabbit knee were cultured by enzyme digestion. Twenty-five μL all-trans-retinoic acid (ATRA) (1×10-6 mol/L) were added in the media of cultured chondrocyte for 24 hours as experimental group, while 25 μL DMEM were added as control group. The secretion of collagen Ⅱ was observed by immunohistochemistry method, cell apoptosis was detected by flow cytometry, IGF-2 mRNA and protein expression in chondrocyte were detected by RT-PCR and Western blot analysis. Results The expression of collagen Ⅱ was down-regulated by ATRA in the experimental group. The cell apoptosis in chondrocyte exposed to ATRA at 1 ×10-6 mol/L was 21% ± 2%, which increased 5 times compared with the control group（5% ± 1%）. The IGF-2 mRNA and protein level in the experimental group were decreased 75% and 57%, respectively, compared to the control group. There weresignificant difference between the experimental group and control group in each index (P lt; 0.05). Conclusion RA may down-regulate the secretion and cell prol iferation, but up-regulate the cell apoptosis in chondrocyte. The apoptotic effect may carry out through inhibiting the IGF-2 expression of chondrocyte.
Objective
To further investigate pathologic mechanism of retinal phototrauma.
Methods
Twenty Wistar rats were divided into control and experimental groups.Their eyes were extracted in 12，24 and 36 hours after light exposure.HE stained retina samples were examined and TDT-mediated dUTP nick end labelling(TUNEL)method was employed to distinguish apoptotic cells.
Results
After 12-hour light exposure,slight vesiculation was observed in the rod outer segment of the retinas.After 24-hour light exposure,the outer nuclear layer showed predominant fractured and condensed nuclei and fragmented DNA.After 36-hour light exposure,the rod outer and inner segments were lysed and most of the nuclei in the outer nuclear layer were disappeared.
Conclusions
Apoptosis of photoreceptor cell is one of the important mechanisms which cause experimental retinal photoinjury of rats.
(Chin J Ocul Fundus Dis, 1999, 15: 167-169)
OBJECTIVE: To investigate apoptosis of chondrocytes cultured in vitro and related expression of caspase-3. METHODS: Apoptosis of chondrocytes were detected by flow cytometry analysis and TUNEL staining. The expression of caspase-3 was determined by RT-PCR and Western blot, and caspase-3 protein activity was determined by ELISA. RESULTS: Apoptosis was observed in chondrocytes cultured in vitro from passage 1 to passage 4 at various degrees. The percentage of apoptosis of chondrocytes on day 7 was much higher than that on day 3 (15.7% +/- 0.3% vs 8.9% +/- 0.6%, P lt; 0.01). caspase-3 mRNA and protein expressed in chondrocytes during whole culture process. Along with the culture time extension in vitro, caspase-3 expression and protein activity up-regulated, coincident with apoptosis of chondrocyte. caspase-3 was activated and a fragment of 20 kDa was detected after 7 days of culture. CONCLUSION: caspase-3 is involved in apoptosis of chondrocytes cultured in vitro.
Objective To observe the expression of cyclin dependent kinase 5 (Cdk5) and p25 in the pathogenesis of retinitis pigmentosa (RP) in Royal College of Surgeon (RCS) rats and its relationships with apoptosis. To explore the mechanism of Cdk5 and p25 induced photoreceptor apoptosis in the pathogenesis of RP. Methods Retinas of RCS and RCS-rdy+ rats were obtained at the ages of postnatal day 17, 25, 35, 60. The retinal structure and thickness of outer nuclear layer were measured by optical microscopy. The expression of Cdk5, p25, cleave-caspase 3 in the retina was evaluated by immunohistochemistry. The protein expression of cleave-caspase 3 in the retina was determined by Western blot. The apoptosis of retinal cells was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The mean absorbance value of apoptotic cells was analyzed by SPSS 17.0 software. Results The retinal thickness of the RCS rats was significantly reduced in comparison to the RCS-rdy+ rats as the postnatal days progressed, particularly in the layer of rods and cones and the outer nuclear layer. The expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increased from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above indexes in RCS-rdy+ rats. The protein expression of cleave-caspase 3 in the RCS rats was significantly increased with progression of postnatal days to postnatal 35; but there was no obvious similar change in RCS-rdy+ rats. The results of TUNEL showed that the apoptotic cells significantly increased in the outer nuclear layer of RCS rats from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above index in RCS-rdy+ rats. This study showed that there were significant correlations between the following variables: Cdk5 expression and p25 expression, Cdk5 expression and cleave-caspase 3 expression, Cdk5 expression and apoptotic cells, p25 expression and cleave-caspase 3 expression, p25 expression and apoptotic cells, cleave-caspase 3 expression and apoptotic cells. The partial correlation coefficients were 0.949, 0.808, 0.959, 0.887, 0.979, 0.852, respectively and the P value was 0.000. Conclusions The apoptotic cells significantly increases and the expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increases from postnatal 17 days to postnatal 35 days. The tendency of apoptotic cells to increase is consistent with the change of Cdk5, p25, cleave-caspase 3 expression. The apoptosis of photoreceptor cells is related to increasing expression of Cdk5 and p25 in RCS rats. Cdk5 may be involved in the development of RP in RCS rats.
Abstract： Stem cell paracrine has been considered as the main mechanism to promote infarcted myocardium regeneration and repair of damaged cardiomyocytes. With further research, secreted frizzled-related protein 2 （Sfrp 2） and stem cell paracrine are closely linked to each other. Sfrp 2 can competitively bind to the specific receptor Fz in Wnt signaling pathway, inhibit Wnt signaling pathway to regulates apoptosis, differentiation, and other life processes of stem cells, and therefore becomes a research hotspot in recent years. This review focuses on the mechanism of Sfrp 2/Wnt signal way in stem cell therapy for myocardial infarction.
Objective To investigate the inhibitory effects of RNA interference (RNAi) expression vector on the expression of survivin in pancreatic cancer cell PANC-1. Methods The protein and mRNA expressions of survivin were examined with immunofluorescence and RT-PCR. The survivin gene was cloned into the T-vector and sequenced. The RNAi expression vectors targeting survivin, named si-svv-1 and si-svv-2 respectively according to whether they harbored a mutation or no mutation, were constructed and transfected into PANC-1 cells with liposome. The expression of survivin mRNA was detected with RT-PCR. Apoptosis of PANC-1 cells was analyzed with DNA ladder and FACS. Results There was a high degree expression of survivin in PANC-1 cells. The expression of survivin was not inhibited by RNAi expression vectors si-svv-1, but inhibited about (72.43±8.04)% by si-svv-2 and the apoptosis rate of PANC-1 cells increased to （12.36±1.44）% after 72 h. Conclusion The RNAi expression vector can effectively inhibit the expression of survivin in pancreatic cancer cell PANC-1 cells and induce the apoptosis in PANC-1 cells.
To elucidate bcl-2 protein expression in hepatic carcinogenetic process and its relationship with apoptotic changes. bcl-2 protein was evaluated immunohistochemically while apoptosis was approached with terminal deoxynucleotidyl transferase (TdT)mediated dUTP nick end labeling (TUNEL) technique in 8 normal livers (NL), 17 liver cirrhosises (LC) and 77 hepatocellular carcinomas (HCC). The results showed that bcl-2 protein was expressed in 3 of 8 NLs(37.5%), 5 of 17 LCs(29.4%) and 7 of 77 HCCs(9.1%) with significant differences between group NL and HCC and between LC and HCC (Ｐ＜0.05). Apoptosis rates of 1.18±0.42%, 4.85±2.78%, 12.89±2.33% in NL, LC, HCC group respectively were demonstrated with significant differences among them (Ｐ＜0.01). Compared the bcl-2 expression with the apoptosis rate in this hepatocarcinogenetic process, reversed trends were presented. Conclusion: bcl-2 expression could be detected in NL, LC and HCC, and its decreasing expression was related to the inhibition attenuation of hepatocellular apoptosis in the process of hepatocarcinogenesis.
OBJECTIVE To study the influence and mechanism of gamma-IFN on fibroblasts in hypertrophic scars(HTS). METHODS The cultured fibroblastic cells were isolated from the hypertrophic scars of 10 patients. The fibroblasts were divided into two groups, one group was treated with gamma-IFN (100 U/ml, 5 days) and the other without gamma-IFN as control. The proliferative activity in both groups was investigated and compared by blood cytometer, the proportion of myofibroblast (MFB) and the ratio of apoptosis were examined and analysed between two groups by flow cytometry using alpha-smooth muscle actin (alpha-SMA) as marker. RESULTS The proliferative activity was downregulated with gamma-IFN. In gamma-IFN treated group, the differentiation of MFB were reduced and the decreasing ratio was 3.2% at the 2nd day and up to 10.5% at the 8th day, then it reduced gradually. The apoptosic ratio is 17.7% in gamma-IFN treated group, and is 10.9% in control group. The difference was statistically significant. CONCLUSION gamma-IFN could downregulate the proliferation of fibroblasts, decrease the differentiation of MFB and induce the apoptosis. It has beneficial effect in the treatment of hypertrophic scars(HTS).
Objective To research the effect of γ-radiation released from 103Pd radioactive stent on the expression of Fas gene and its relation with apoptosis of bile duct cancer cells lines. Methods The cancer cells of bile duct were dissociated into suspension in culture flasks, and the number of cells was counted by hemacytometry. The suspension was then stored in 2 ml freezing tubes in the density of 1×105/ml. They were set in two stents: general stent （general stent group） and 103Pd radioactive stent (103Pd stent group). The expression of Fas gene and apoptosis of bile duct cancer cells in general stent group and 103Pd stent group were analysed with immunohistochemistry technique and TUNEL method, respectively. Results The expression level of Fas gene in 103Pd stent group was significantly higher than that in general stent group （P＜0.05）, and the number of apoptotic cancer cells in 103Pd stent group was also significantly more than that in general stent group （P＜0.01）. Conclusion There is a correlation between the expression of Fas gene and the apoptosis of bile duct cancer cells, which means that 103Pd radioactive stent may increase the expression of Fas gene and promote the apoptosis of cancer cells. It may be helpful for the further study of treatment for bile duct cancer using 103Pd radiative stent.
