ABSTRACT
Background: Cervical cancer is known to have a good response to radiotherapy. The response and prognosis are dependent on the level of apoptosis. Pap smear and histopathology are cost-effective methods in diagnosing premalignant and malignant lesions of cervix but not accurate in classifying and estimating the progression of the disease, especially in premalignant lesions. Therefore this study was undertaken to know the role of Ki-67 expression and apoptotic index in classifying accurately the premalignant lesions for better management.Methods: The study included 540 cases diagnosed histologically as cervical intraepithelial neoplasia or carcinoma. The apoptotic index is calculated for all the 540 cases using light microscopy on Haematoxylin and Eosin stained sections. Ki-67 immunohistochemical staining was done for 100 cervical biopsies. Ki-67 expression was graded and the Ki-67 labelling index was calculated. Statistical evaluation was done using the unpaired t-test.Results: The Apoptotic index increased with increasing grade of dysplasia. There is a significant difference in the mean apoptotic index between premalignant and malignant lesions of the cervix. The ki-67 index increased with increasing grade of dysplasia. There is a significant difference in the mean Ki-67 index between premalignant and malignant lesions of the cervix.Conclusions: Apoptotic index and proliferative indices have been found useful in distinguishing between premalignant and malignant lesions of the cervix and gives an idea about the proliferative activity of the tumour for better management of the patient and to determine prognosis.
ABSTRACT
BACKGROUND: To date, it has been confirmed that etomidate pre-treatment can reduce the damage of remote organs caused by limb ischemia-reperfusion, but whether etomidate post-treatment has protective effect on remote organs and its mechanism has been rarely reported. OBJECTIVE: To investigate the influence of etomidate post-treatment on limb ischemia-reperfusion lung injury. METHODS: A rat model of limb ischemia-reperfusion lung injury was prepared by clamping the bilateral femoral arteries for 2 hours and reperfusion for 3 hours. After 2 hours of limb ischemia, I/R group experienced the process of limb ischemia-reperfusion; I/R+ETO group, I/R+Dex 0.2 group, I/R+Dex 0.5 group and I/R+Dex 1.0 group, besides the model of limb ischemia-reperfusion, were injected with etomidate 1.0 mg/kg and dexamethasone 0.2, 0.5 and 1.0 mg/kg respectively through tail vein. At 3 hours of reperfusion, blood samples were extracted from the carotid artery, blood gas analysis was performed and the partial pressure of blood oxygen (PaO2) was recorded. The pathological changes were detected by immunohistochemistry. Apoptotic index was detected by Hoechst 33258 staining and wet/dry weight ratio was detected. Fas protein and Fasl mRNA of lung tissue were detected by western blot and RT-PCR respectively. Tumor necrosis factor-α and interleukin-1β levels were detected by ELISA. RESULTS AND CONCLUSION: Compared with the I/R group, PaO2 increased (P 0.05). To conclude, etomidate post-treatment can reduce lung injury caused by limb ischemia-reperfusion in rats, and its mechanism may be related to the down-regulation of Fas/FasL. In the statistical point of view, etomidate 1.0 mg/kg has the potency intensity of reducing lung injury, almost equivalent to dexamethasone 0.5 mg/kg.
ABSTRACT
Objective@#To investigate the effect of Ethephon on the testis pathological structure and apoptosis of spermatogenic cell in offspring male rats.@*Methods@#Twenty-four healthy female SD rats of 45 days old were randomly divided into control group, low-dose Ethephon group, medium-dose Ethephon group and high-dose Ethephon group according to body weight.The male rats of the same age were selected to mate with female rats.The rats were fed with Ethephon solution of different concentrations or 9 g/L saline every day, and they were continued to be fed with Ethephon during pregnancy and lactation.At the age of 7 days and 14 days, 10 offspring male rats were randomly selec-ted from each group and were put to death.The testicular tissue was stained with HE, and the morphological changes in the testis were observed with light microscope; the apoptotic cells were labeled with terminal deoxynucleotidyl transfe-rase dUTP nick-end labeling (TUNEL method) and the apoptosis index(AI) of testis spermatogenic cells was detected with fluorescence microscope.@*Results@#At the age of 7 days, the testis internal structure of the control group developed well, and the spermatic tubules were neatly and compactly arranged.In the low-dose Ethephon group, the seminiferous tubules of the testis were slightly smaller and the spermatogenic cells were loosely arranged compared with the control group.In the medium-dose Ethephon group, the testis seminiferous tubules were slightly disordered and the cell gap increased.In the high-dose Ethephon group, the testis development was poor, the diameter of seminiferous tubules decreased significantly, and the spermatogenic cells arrangement was in disorder.There was no statistically significant difference in spermatogenic cell AI between the low-dose group [(0.54±0.10)%] and the control group[(0.53±0.09)%] (P>0.05), while the spermatogenic cell AI in the medium-dose Ethephon group [(0.63±0.11)%] and the high-dose Ethephon group [(0.81±0.06)%] were higher than that in the control group, thus there exists a statistically significant difference (all P<0.01). The spermatogenic cell AI in the low-dose Ethephon group [(0.54±0.10)%] was lower than that in the medium-dose Ethephon group [(0.63±0.11)%], and the difference was statistically significant (P<0.05). The spermatogenic cell AI in the medium-dose Ethephon group was higher than that in the high-dose Ethephon group, and the difference was statistically significant (P<0.01). At the age of 14 days, the spermatogenic cells AI in control group, low-dose Ethephon group, medium-dose Ethephon group and high-dose Ethephon group were (0.54±0.08)%, (0.65±0.11)%, (0.77±0.11)%, and (0.88±0.10)% respectively, and the spermatogenic cells AI in all groups increased gradually, in which the differences were statistically significant (all P<0.01).@*Conclusions@#Excessive dose of Ethephon can induce pathological changes in testicular tissue and increase the apoptosis of spermatogenic cells, resulting in low fertility of offspring rats.
ABSTRACT
Objective To investigate the effect of Ethephon on the testis pathological structure and apoptosis of spermatogenic cell in offspring male rats.Methods Twenty-four healthy female SD rats of 45 days old were randomly divided into control group,low-dose Ethephon group,medium-dose Ethephon group and high-dose Ethephon group according to body weight.The male rats of the same age were selected to mate with female rats.The rats were fed with Ethephon solution of different concentrations or 9 g/L saline every day,and they were continued to be fed with Ethephon during pregnancy and lactation.At the age of 7 days and 14 days,10 offspring male rats were randomly selected from each group and were put to death.The testicular tissue was stained with HE,and the morphological changes in the testis were observed with light microscope;the apoptotic cells were labeled with terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL method) and the apoptosis index(AI) of testis spermatogenic cells was detected with fluorescence microscope.Results At the age of 7 days,the testis internal structure of the control group developed well,and the spermatic tubules were neatly and compactly arranged.In the low-dose Ethephon group,the seminiferous tubules of the testis were slightly smaller and the spermatogenic cells were loosely arranged compared with the control group.In the medium-dose Ethephon group,the testis seminiferous tubules were slightly disordered and the cell gap increased.In the high-dose Ethephon group,the testis development was poor,the diameter of seminiferous tubules decreased significantly,and the spermatogenic cells arrangement was in disorder.There was no statistically significant difference in spermatogenic cell AI between the low-dose group [(0.54 ± 0.10)％] and the control group[(0.53 ±0.09) ％] (P ＞ 0.05),while the spermatogenic cell AI in the medium-dose Ethephon group [(0.63 ± 0.11) ％]and the high-dose Ethephon group [(0.81 ± 0.06) ％] were higher than that in the control group,thus there exists a statistically significant difference (all P ＜0.01).The spermatogenic cell AI in the low-dose Ethephon group [(0.54 ±0.10) ％] was lower than that in the medium-dose Ethephon group [(0.63 ± 0.11)％],and the difference was statistically significant (P ＜ 0.05).The spermatogenic cell AI in the medium-dose Ethephon group was higher than that in the high-dose Ethephon group,and the difference was statistically significant (P ＜0.01).At the age of 14 days,the spermatogenic cells AI in control group,low-dose Ethephon group,medium-dose Ethephon group and high-dose Ethephon group were (0.54 ± 0.08) ％,(0.65 ± 0.11) ％,(0.77 ± 0.11) ％,and (0.88 ± 0.10) ％respectively,and the spermatogenic cells AI in all groups increased gradually,in which the differences were statistically significant (all P ＜ 0.01).Conclusions Excessive dose of Ethephon can induce pathological changes in testicular tissue and increase the apoptosis of spermatogenic cells,resulting in low fertility of offspring rats.
ABSTRACT
Background & objectives: Significance of apoptosis as a prognostic marker is less well studied in paediatric acute lymphoblastic leukaemia (ALL) cases. Hence, a prospective study, involving 30 paediatric ALL cases, was done to assess the clinical relevance of in vivo apoptosis. Methods: Peripheral blood mononuclear cells from all patients were subjected to annexin V/propidium iodide staining to detect the degree of apoptosis [apoptotic index (AI)] at day 0 and day 35 post-induction chemotherapy. In addition, Bax and Bcl2 apoptotic protein expressions were studied at day 0 and their relative fluorescence mean intensity (RFMI) ratios were calculated. Results: Mean age of patients was 5.1 years. Of the 30 cases, 21 (70%) were at standard-risk, five (17%) at intermediate and four (13%) at high risk. Majority (83%) were B-ALL. Day 8 absolute blast count was >1000/?l in seven (23%) and <1000/?l in 23 of 30 (77%) cases. Day 35 marrow was M1 in 23 (92%) and M2 in two of 25 (8%) cases. AI at day 0 and day 35 ranged from 0.9 to16.6 per cent and 1.4 to 62.8 per cent with a mean of 5.90 and 19.64 per cent, respectively. The Bax/Bcl2 ratio ranged from 0.2 to 3.5 with a mean of 0.83. The ratio was predominantly anti-apoptotic, i.e. <1 (77%). A significant association was noted between low AI at day 0 and high total leucocyte count (P=0.02), T-cell phenotype (P=0.043) and high-risk as per NCI category (P=0.025). Significant increase (>30%) in day 35 AI was seen in only six cases. Interpretation & conclusions: Our study showed that low AI at day 0 was associated with a high-risk clinical phenotype in paediatric ALL. However, studies on larger group, especially with longer follow up or study of relapse cases, will help draw conclusions regarding apoptosis assessment in paediatric ALL.
ABSTRACT
Objective: To investigate the protective effect of Qijudihuang Pill on the retinal structures of the glaucoma model rats, and to clarify its mechanism. Methods: Fifty SD rats were randomly divided into blank control group, model group, and low, medium, and high doses of Qijudihuang Pill groups (n=10). The chronic glaucoma rat models were established by using cauterization on episcleral veins in each group except blank control group. The rats in low, medium, and high doses of Qijudihuang Pill groups were given 1, 2, and 4 g · kg-1 Qijudihuang Pill by gavage, while the rats in blank control group and model group were given the same volume of saline, 1 time per day. After 12 weeks of administration, the morphological changes in ocular tissue were analyzed by HE staining. The apoptosis of retinal ganglion cells was detected by TUNEL staining. The expression levels of Bel-2, Bax, and caspase-3 mRNA in the retina tissue of the rats in various groups were determined by RT-PCR. The expression levels of Bel-2, Bax, and caspase-3 proteins in the retina tissue of the rats in various groups were determined by immunohistochemistry. Results; Compared with blank control group, the retinal nerve fibers in model group were irregularly arranged, the interfibrillar substance was edematous, the cell layer was vacuolated, and the number of retinal ganglion cells was reduced significantly. Compared with model group, the number of retinal ganglion cells of the rats in high dose of Qijudihuang Pill group was increased; the retinal nerve fiber cells in the eyes of the rats in low dose of Qijudihuang Pill group and medium dose of Qijudihuang Pill group were arranged neatly, and the morphology of ganglion cells were normal. Compared with model group, the apoptotic indexes of retinal ganglion cells in low, medium, and high doses of Qijudihuang Pill groups were decreased significantly (P<0. 05); the mRNA and protein expression levels of Bcl-2 were increased significantly (P <0. 05), while the mRNA and protein expression levels of Bax and caspase-3 were decreased significantly (P<0. 05). Conclusion: Qijudihuang Pill can protect the retina of glaucoma rats by regulating the mRNA and protein expressions of Bcl-2, Bax, and caspase-3 and inhibiting the apoptosis of retinal ganglion cells.
ABSTRACT
Background: Apoptotic index (AI) using light microscopy as an indirect measure to assess the significance of apoptosis as a proliferative marker in dysplastic lesions and malignant epithelial lesions of the oral cavity. Aims: (1) To quantify the apoptotic bodies/cells in oral epithelial dysplastic (OED) lesions and oral squamous cell carcinoma (OSCC). (2) To measure AI in OED and OSCC. (3) To compare AI in OED and OSCC. Settings and Design: The proposed laboratory‑based retrospective study involved the use of hematoxylin and eosin (H and E)‑stained slides of previously diagnosed OED lesions and OSCC from institutional archives. Materials and Methods: This study constituted 50 cases, each of H and E‑stained slides of previously diagnosed cases of OED and OSCC. AI was calculated as the number of apoptotic bodies/cells expressed as a percentage of the total number of nonapoptotic tumor/dysplastic cells counted in each case. Statistical Analysis Used: Nonparametric tests such as Kruskal–Wallis test and Mann–Whitney test were used. Results: There was a statistically significant increase in AI from OED to OSCC (P = 0.000). Conclusions: Further studies need to be undertaken to detect and understand the apoptotic mechanisms in the progression from OED to OSCC.
ABSTRACT
Objective To explore the apoptosis effect of docetaxel combined gamma knife on hepatoma cell SMMC-7721 subcutaneous xenograft in nude mice.Methods Subcutaneous xenogyaft models were constructed and were divided into two groups:control group and experimental group.The experimental group was treated with docetaxel 60ug/0.3ml once every 3 days for 6 times and gamma irradiation once every other day for 6 times (with indoor temperature of 137Cs radiation source irradiating the tumor and of fractionated schedule 5Gy with the total dose of 10Gy every time).The control group was treated with physiological saline with the same dose of 60 ug/0.3 mL.Tumor growth was observed.Tumor samples were cut 30 days after the treatment and TUNEL was used to detect the apoptosis of tumor cells.Results Tumor growth rate in experimental group significantly slowed down.Apoptotic index in experimental groups was significantly higher than that in control group (P ＜ 0.05) Cornclusion Docetaxel combined gamma knife can inhibit the growth of hepatocellular carcinoma in nude mice.
ABSTRACT
Apoptosis is a process of programmed cell death occurring in multicellular organisms in whom development, maintenance and sculpturing organs and tissues. Taken together, apoptotic processes are of widespread biological significance; being involved in e.g. development, differentiation, proliferation/homoeostasis, regulation and function of the immune system and in the removal of defected harmful cells. Dys regulation of apoptosis can play a primary or secondary role leading to cancer whereas excessive apoptosis contributes to neuro degeneration, autoimmunity, AIDS, and ischemia. Gaining insight into the techniques for detecting apoptotic cells will allow the development of more effective, higher specific and therefore better-tolerable therapeutic approaches. The goal of this review article is to provide a general overview of current knowledge, on the various technical approaches for detecting apoptotic cells.
Subject(s)
Apoptosis , Comet Assay/methods , DNA Fragmentation , Electrophoresis/methods , Flow Cytometry/methods , Humans , Immunohistochemistry/methods , In Situ Nick-End Labeling/methods , Microscopy/methodsABSTRACT
Aim This study explored the expression of HIF-1α in hypoxic cardiac muscle in mice, and observed the evidence of apoptosis in hypoxia induced cardiomyocyte. Methods Male Sprague-Dawley rats, were randomized into 7 groups (n= 4 per group): control normoxia group that was exposed to atmospheric oxygen and hypoxia groups that were housed in hypoxic chambers (O2 level 8%) for 1, 3, 7, 14, 21, and 28 days respectively. Animals were sacrificed, hearts were rapidly excised, total RNA was extracted with an mRNA isolation kit and the expression of HIF-1α mRNA was then detected by real-time RT-PCR. Apoptosis was assessed by TUNEL method. Results For rat in hypoxia group, the expression of HIF-1α mRNA in cardiac myocytes was clearly up-regulated compared to the control normoxia group. Further, HIF-1α expression level elevated gradually and reached a peak at 21 days of hypoxia. No cell labeled by the TUNEL method was detected in the control group. Compared with the control group, the apoptotic index was significantly increased in the hypoxia group (P < 0.05). There was no significant correlation between the elevation of HIF-1α mRNA and the elevation of apoptotic index. Conclusion Systemic chronic hypoxia caused the elevation of HIF-1α mRNA and apoptosis in cardiac myocytes.
Subject(s)
Rats, Sprague-Dawley , Myocardium , ApoptosisABSTRACT
OBJECTIVE: To determine the effects of neoadjuvant chemotherapy (5-fluorouracil plus cisplatin) on tumor cell morphology and apoptosis by analyzing the consecutive changes of apoptotic index (AI) and histology observed in the serially obtained cervical cancer tissues during the chemotherapy. METHODS: Cervical cancer tissues were obtained by punch biopsy just before starting the each cycle of neoadjuvant chemotherapy from five patients with locally advanced disease (stage IIb-IIIb), but previously untreated squamous cell carcinoma of uterine cervix. All patients were treated with three cycles of 5-fluorouracil (1,000 mg/m2 at day #1-5) and cisplatin (60 mg/m2 at day #1) at 3 weeks interval. All H & E stained cervical cancer tissue slides were scored for apoptotic index and observed for microscopic changes of tumor cells by a pathologist. RESULTS: After the first cycle of chemotherapy, AI was significantly increased (from 2 times to 8 times). And widespread injury to cytoplasm was observed and followed by karyorrhexis and karyolysis of nucleus of tumor cells. The size of tumor nests was reduced and it was also noted that fibrosis and infiltration of inflammatory cells were increased. The parts of tumor nests were replaced by mature squamous cells and the changes in nuclear morphologic features pointing in a more differentiated direction. But after the second cycle of chemotherapy, only one patient showed an increase in AI by 1.2 times over that after the first cycle of chemotherapy. The rest showed slight decreases in AI compared to that after the first cycle of chemotherapy. In addition, fewer microscopic morphologic changes of tumor cells induced by chemotherapy were observed after the second cycle of chemotherapy compared to those after the first cycle of chemotherapy. CONCLUSION: We found that AI hardly increased or rather decreased, and that microscopic changes of tumor cells were fewer after the second cycle of neoadjuvant chemotherapy compared to the situation after the first cycle of chemotherapy. Thus, we could deduce that chemoresistance might rapidly develop in cervical cancer cells after the first cycle of neoadjuvant chemotherapy using 5-fluorouracil and cisplatin. So we need to consider this problem when we treat the locally advanced cervical cancer patients with neoadjuvant chemotherapy using 5-fluorouracil and cisplatin.
Subject(s)
Female , Humans , Apoptosis , Biopsy , Carcinoma, Squamous Cell , Cervix Uteri , Cisplatin , Cytoplasm , Drug Therapy , Fibrosis , Fluorouracil , Uterine Cervical NeoplasmsABSTRACT
BACKGROUND: Keratoacanthoma (KA) is a unique neoplasm, usually accompanied by rapid growth and regression, and the histologic findings resemble those of squamous cell carcinoma (SCC). Histologic differentiation of KA and SCC is challenging, therefore a number of studies have been carried out to differentiate these two entities using immunohistochemical staining. However, the results were inconsistent. There is now a debate as to whether KA is a benign tumor or a less aggressive variant of SCC. OBJECTIVE: The purpose of this study was to examine the expression patterns of MMP-3, MMP-7, TIMP-1, the apoptotic index in KA and SCC, and the usefulness of immunohistochemical staining in differentiating these two entities. METHODS: Formalin-fixed, paraffin-embedded biopsy specimens obtained from 12 KA and 16 SCC patients were investigated for the expressions of MMP-3, MMP-7 and TIMP-1 using an immnohistochemical staining method and apoptotic index by TUNEL method. RESULTS: 1. The expression of MMP-3 in mass was slightly higher in the SCC group (3.4+/-1.03) than the KA group (2.7+/-1.50). The expression of MMP-3 on the stroma was significantly higher for the SCC group (2.6+/-0.96) than the KA group (1.5+/-1.24) (p<0.05). 2. The expression of MMP-7 was detected only in a single case within the SCC group. 3. The expression of TIMP-1 was detected in two cases within the KA group. 4. The apoptotic index (%) was significantly higher in the SCC group (57.3+/- 32.15) than the KA group (36.1+/-21.78) (p<0.05). CONCLUSION: The immunohistochemical staining method of MMP-3, MMP-7 and TIMP-1 is not sensitive enough to differentiate between the two diseases. However, it has been suggested that MMP-3 of the stroma is involved in invasion of SCC more than MMP-3 of the mass, that MMP-7 plays a role in the early stage of SCC, and that TIMP-1 inhibits the action of MMPs and invasion in KA. We suggest that KA can be regarded as a clinically benign end of the SCC disease spectrum. The apoptotic index was higher in the SCC group than the KA group, implying that SCC has greater invasive ability due to a newly-selected, mutated, neoplastic cellular clone that has more aggressive biological behavior.
Subject(s)
Humans , Apoptosis , Biopsy , Carcinoma, Squamous Cell , Clone Cells , In Situ Nick-End Labeling , Keratoacanthoma , Matrix Metalloproteinases , Tissue Inhibitor of Metalloproteinase-1ABSTRACT
OBJECTIVE: The aim of this study was to identify the relationship between the progression of CIN (cervical intraepithelial neoplasia) to cervical cancer and expression pattern of VEGF, PCNA and apoptosis. METHODS: We prepared forty paraffinized cervical tissue blocks consisting of each 10 CINI, CINII, CINIII, and invasive cervical cancer tissues, which had been already diagnosed histologically in Seoul National University Hospital from July, 1998 to June, 1999. The protein expression of VEGF and PCNA were examined by immunohistochemical staining. We defined PI (positive index) as the percentage of cells that were positive PCNA staining when examined in high power fields with light microscope (x400). We used TUNEL staining to calculate apoptotic index defined as the number of the cells undergoing apoptosis per 1,000 tumor cells. To detect HPV type 16 or 18 from paraffin-embedded tissues, we selected the nested PCR method. RESULTS: The VEGF expression showed significant difference between early cervical lesions (CINI & II) and advanced cervical lesions (CINIII & cancer) (p=0.041). In case of PCNA expression there was no significant difference. When we define positive case as the case of which AI was greater than 5, AI according to each diagnosis showed no difference. In the aspect of HPV infection we could find that there was significant increment of HPV 16/18 infection rate in advanced cervical lesions (CINIII, cervical cancer) comparing with early cervical lesions such as CINI, CINII. CONCLUSION: Our results suggest that the evaluation of VEGF expression and HPV infection is potentially useful for the prediction of the progression of CIN to carcinoma.
Subject(s)
Apoptosis , Diagnosis , In Situ Nick-End Labeling , Paraffin , Polymerase Chain Reaction , Proliferating Cell Nuclear Antigen , Seoul , Uterine Cervical Neoplasms , Vascular Endothelial Growth Factor AABSTRACT
PURPOSE: Tumor growth in a given neoplasm is the net result of cell proliferation and cell loss, and apoptosis is the most significant component of continuous cell loss in most tumors. In this study, we examined non-Hodgkin's lymphoma (NHL, n=67) immunohistochemically for the presence of Bcl-2 oncoprotein and P53 protein and compared apoptotic indices (AIs) and Ki-67 proliferative indices (percentages of Ki-67 positive cells). MATERIALS AND METHODS: 67 patients with NHL were evaluated : 3 low-grade and 64 intermediate-grade. The phenotype was determined in 65 cases : 47 (70%) were B cell type and 18 (27%) were T cell type. AIs and Ki-67 proliferative indices were determined immunohistochemically and the overexpression of P53 and Bcl-2 protein were also evalutated. RESULTS: The overexpressions of Bcl-2 protein and P53 protein were found in 40% (26/65) and 31% (20/ 65). The AI ranged from 0% to 15% (mean 2.16, median 1.2). Cellular Bcl-2, which counteracts apoptosis, was significantly ( p=0.005) associated with AIs. Ki-67 proliferative indices ranged from 1% to 91% (mean 55.4), and P53 was significantly ( p=0.000) associated with Ki-67 proliferative indices. A positive correlation between AIs and Ki-67 proliferative indices was revealed ( p=0.012) in Bcl-2 positive patients. CONCLUSION: In NHL, we observed a correlation between AIs and Bcl-2 expression, between Ki-67 proliferative indices and P53 expression, and between AIs and Ki-67 proliferative indices in Bcl-2 positive patients. Our results suggest that cell apoptosis may be inseparable from cell proliferation during tumor growth.
Subject(s)
Humans , Apoptosis , Cell Proliferation , Lymphoma, Non-Hodgkin , PhenotypeABSTRACT
Background and purpose:Members of the NF-kappaB family of transcription factors could activate bcl-2 anti-apoptotic genes at the transcriptional level and then regulate cell apoptosis.The p53 tumor suppressor gene has a clear role!in both the regulation of cell cycle and expression of apoptosis-related bcl-2 family.There were few reports about the expression of NF-?Bp65,bcl-2,bcl-xL and their relationships with p53 and cell apoptosis in pancreatic cancer.The present study was designed to analyze the expression of the antiapoptotic molecules nuclear factor kappaB(NF-?Bp65),bcl-2 and bcl-xL in pancreatic ductal carcinoma(PC) and their correlations to the apoptosis index and p53 expression.Methods:Immunohistochemical analyses of P53 protein was performed on 25 cases of pancreatic ductal carcinoma and 9 cases of normal pancreas;Western blot was used to evaluate NF-?Bp65 protein and RT-PCR for bcl-2 mRNA and bcl-xL mRNA of 25 cases of pancreatic ductal carcinoma and 9 NP cases.The apoptosis was determined by TUNEL.Results:The positive rate of P53 protein was 56%(14/25)in PC tissues,significantly higher than that in NP tissues(P
ABSTRACT
Apoptosis, including the programmed cell death, is important event in normal cell turnover and maintenance of adult tissues. Apoptosis exerts a homeostatic function in relation to tissues dynamics, as the steady state of continuously renewing tissues achieved by a balance between cell replication and cell death. This study was undertaken to investigate the association between apoptosis and development of the cervical neoplasia. Archival cervical samples from normal epithelium (n 10), low-grade squamous intraepithelial lesions (LSIL, n = 10), high-grade squamous intraepithelial lesions (HSIL, n 10), microinvasive squamous cell carcinomas (n 10), and invasive squamous cell carcinomas (n = 10) were evaluated for apoptosis. We used in situ end-labeling of DNA strand breaks by terminal deoxynucleotidyltransferase incorporation of biotinylated deoxyuridine to 3-OH ends of DNA, identified by nickel-avidine-peroxidase. The apoptotic index (sum of apoptotic bodies divided by the total nuclei times 100) significantly decreased (P<0.05) as the degree of neoplasia increased: 3.1 + 0.9 % in normal epithelium, 5.5 +/- 1.4 % in LSIL, 1.6 +/- 0.4 % in HSIL, 1.9 +/- 0.5 % in microinvasive carcinomas, and 0.6 +/- 0.3 % in invasive carcinomas. Compared to normal epithelium, the total cell number per 200x field increased significantly (P<0,05): 379 +/- 47 in normal epithelium, 462 +/- 228 in LSIL, 670+/-293 in HSIL, 1035 +/- 254 in microinvasive carcinomas, and 1389 +/- 247 in invasive carcinomas. Consequently, these results suggest that progession of cervical carcinogenesis is associated with a decrease in apoptotic index and an increase in the number of the total cell.
Subject(s)
Adult , Humans , Apoptosis , Carcinogenesis , Carcinoma, Squamous Cell , Cell Count , Cell Death , Deoxyuridine , DNA , DNA Nucleotidylexotransferase , EpitheliumABSTRACT
p27kip1, a cyclin dependent kinase inhibitor, has been recognized as a negative regulator of cell cycle. To investigate the role of p27kip1 on progression of cancer and apoptotic pathway, we analyzed p27kip1 expression using immunohistochemical stain in 40 cases of prostatic adenocarcinoma and apoptotic index by TUNEL method in 30 cases of prostatic adenocarinoma. Both were correlated with Gleason grade and Gleason score. Loss of p27kip1 expression was more frequent in prostatic adenocarcinomas of higher score (Gleason score 7 to 10) (60.7%) than in those of lower score (Gleason score 4 to 6) (33.3%) (p0.05). These results suggest that loss of p27kip1 expression and increased apoptotic index may be the morphologic markers to predict the behavior of prostatic adenocaricnoma. The role of p27kip1 on apoptotic pathway seems to be meager in this study and needs further study.
Subject(s)
Adenocarcinoma , Cell Cycle , Cyclins , In Situ Nick-End Labeling , Neoplasm Grading , PhosphotransferasesABSTRACT
Apoptosis is a distinct mode of cell death that is responsible for deletion of cells in normal tissues. Apoptotic cell death plays an important role in the proliferation and turnover of cells in various tumors. Apoptosis occurs spontaneously in malignant tumors, often markedly retarding their growth, and increased in tumors responding to irradiation, cytotoxic chemotherapy, heating and hormone ablation. Flowcytometric analysis of the cellular DNA content appears to be a useful clinical prognostic indicator in colorectal cancer. The relationship of apoptotic index(AI) and proliferative indices have being investigated. We analyzed the tumor DNA content and AI in 84 patients who underwent resection for colorectal cancer between January 1989 and December 1994 in order to evaluate the prognostic significance of apoptosis, DNA ploidy and index using in situ apoptosis detection method and flowcytometry. The mean value of AI was 32.4, and median value 21. In the cellular DNA, forty-two percent of the tumors were diploidy, fifty-eight percent aneuploidy. The mean value of DNA index(DI) was 1.38, G0/G1 72%, S phase fraction 21.7%, G2/M 6.3%, and proliferative fraction 28%. There was no significant difference between AI and tumor invasion, LN metastasis, DNA ploidy, DI.(p>0.05) There was no significance between overall survival and AI, DNA ploidy, DI. But patients who had tumors with low DNA index had a significantly longer disease free survival than high DNA index.(p<0.05) As a result, this study shows that AI is a less useful as prognostic factor and DNA index is a more important prognostic factor in patients undergoing surgery for colorectal cancer.