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1.
Chinese Critical Care Medicine ; (12): 921-926, 2022.
Article in Chinese | WPRIM | ID: wpr-956077

ABSTRACT

Objective:To explore the value of monocyte subsets and CD64 expression in the diagnosis and prognosis of sepsis.Methods:A prospective case-control study was designed. 30 septic patients and 30 non-septic patients who were admitted to the intensive care unit (ICU) of the PLA Army Characteristic Medical Center from March 2021 to March 2022 were enrolled. After 1, 3, and 5 days of ICU admission, peripheral blood samples were taken from patients. Flow cytometry was used to detect the proportion of monocyte subsets and the expression level of CD64 on the surface, and the difference of expression between patients in two group was analyzed. The risk variables for sepsis were analyzed using single-factor and multi-factor Logistic regression. The diagnostic efficacy of each risk factor for sepsis was determined using the receiver operator characteristic curve (ROC curve).Results:One day after ICU admission, the proportions of monocytes and classic monocytes in white blood cells (WBC) of septic patients were significantly lower than those of non-septic patients [proportion of monocytes to WBC: (4.13±2.03)% vs. (6.53±3.90)%, proportion of classic monocytes to WBC: 1.97 (1.43, 2.83)% vs. 3.37 (1.71, 5.98)%, both P < 0.05]. The proportion of non-classical monocytes in monocytes was significantly higher in septic patients than that in non-septic patients [(11.42±9.19)% vs. (6.57±4.23)%, P < 0.05]. The levels of CD64 expression in monocytes, classic monocytes, intermediate monocytes and non-classic monocytes were significantly higher in sepsis patients than those in non-septic patients [mean fluorescence intensity (MFI): 13.10±6.01 vs. 9.84±2.83 for monocytes, 13.58±5.98 vs. 10.03±2.84 for classic monocytes, 13.48±6.35 vs. 10.22±2.99 for intermediate monocytes, 8.21±5.52 vs. 5.79±2.67 for non-classic monocytes, all P < 0.05]. Multivariate Logistic regression research showed that CD64 in typical monocytes [odds ratio ( OR) = 1.299, 95% confidence interval (95% CI) was 1.027-1.471, P = 0.025] and the proportion of non-typical monocytes in monocytes ( OR = 1.348, 95% CI was 1.034-1.758, P = 0.027) were the independent risk factors for sepsis. ROC curve showed that the area under the ROC curve (AUC) of CD64 expression of classical monocytes, the fraction of non-classical monocytes in monocytes, and procalcitonin (PCT) in the diagnosis of sepsis was 0.871. A correlation analysis revealed a negative relationship between the acute physiology and chronic health status evaluation Ⅱ (APACHE Ⅱ) on the first, third, and fifth days following ICU admission and the expression level of CD64 in patients' classic monocytes ( r values were -0.264, -0.428 and -0.368, respectively, all P < 0.05). Conclusions:Combining the proportion of non-classical monocytes in monocytes, the level of plasma PCT, and the CD64 expression of classic monocytes in peripheral blood has good efficacy in identifying sepsis and assessing its severity.

2.
Chinese Critical Care Medicine ; (12): 223-228, 2021.
Article in Chinese | WPRIM | ID: wpr-883862

ABSTRACT

Objective:To study the dynamic changes of cellular immune function in peripheral blood of trauma patients and its role in the evaluation of traumatic complications.Methods:A prospective cohort study design was conducted. Patients with blunt trauma admitted to Chongqing Emergency Medical Center from November 2019 to January 2020 were consecutively enrolled. The peripheral blood samples were collected at 1, 3, 5, 7, and 14 days after injury. The expressions of CD64, CD274, and CD279 on the surface of neutrophils, lymphocytes, and monocytes as well as CD3 +, CD4 + and CD8 + T lymphocyte subsets were measured by flow cytometry. The trauma patients were divided into different groups according to the injury severity score (ISS) and sepsis within 28 days after injury, respectively. The dynamic changes of cellular immune function in different time points after injury and differences between different groups were compared. Furthermore, the correlation with acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), sequential organ failure assessment (SOFA), and ISS were evaluated by Pearson correlation analysis. Results:A total of 42 patients with trauma were finally enrolled, containing 8 severe trauma patients with ISS greater than 25 scores, 17 patients with ISS between 16 and 25 scores, and 17 patients with ISS less than 16 scores. The sepsis morbidity rates were 14.3% (n = 6) within 28 days after injury. CD64 index and CD4 +T lymphocyte subsets were significantly increased at different time points after trauma (H = 15.464, P = 0.004; F = 2.491, P = 0.035). The CD64 index and positive rates of CD279 in neutrophils, lymphocytes, and monocytes were increased with the severity of injury at day 1 and day 3 after injury, respectively. At the first day after injury, CD64 index were 2.81±1.79, 1.77±0.92, 3.49±1.09; positive rate of CD279 in neutrophils were 1.40% (0.32%, 2.04%), 0.95% (0.44%, 2.70%), 12.73% (3.00%, 25.20%); positive rate of CD279 in lymphocytes were 3.77% (3.04%, 5.15%), 4.71% (4.08%, 6.32%), 8.01% (4.59%, 11.59%); positive rate of CD279 in monocytes were 0.57% (0.24%, 1.09%), 0.85% (0.22%, 1.25%), 6.74% (2.61%, 18.94%) from mild to severe injury groups, respectively. The CD64 index in severe injury group was significantly higher than that in moderate group, and the positive rates of CD279 in neutrophils, lymphocytes and monocytes of severe injury patients were higher than those in other two groups (all P < 0.05). At 3rd day after injury, compared to moderate group, severe injury patients had significantly higher CD64 index and positive rate of CD279 in lymphocytes [4.58±2.41 vs. 2.43±1.68, 7.35% (5.90%, 12.28%) vs. 4.63% (3.26%, 6.06%), both P < 0.05]. Compared with the non-sepsis patients, the sepsis patients had significantly higher CD64 index and positive rate of CD279 in monocytes at day 1 after injury [4.06±1.72 vs. 2.36±1.31, 3.29% (1.14%, 12.84%) vs. 0.67% (0.25%, 1.48%), both P < 0.05], and positive rate of CD279 in lymphocytes significantly higher at 3rd day after injury [8.73% (7.52%, 15.82%) vs. 4.67% (3.82%, 6.21%), P < 0.05]. In addition, correlation analysis showed that positive rate of CD279 in lymphocytes was positively correlated with SOFA and ISS, respectively (r values were 0.533 and 0.394, both P < 0.05), positive rate of CD279 in monocytes was positively correlated with APACHEⅡ, SOFA and ISS scores, respectively (r values were 0.579, 0.452 and 0.490, all P < 0.01), positive rate of CD279 in neutrophils was positively correlated with APACHEⅡ and ISS, respectively (r values were 0.358 and 0.388, both P < 0.05). Conclusions:CD64 index and CD279 expression in neutrophils, lymphocytes, and monocytes are significantly related to the severity and prognosis of trauma. Dynamic monitoring the cellular immune function may be helpful for assessing the prognosis of trauma patients.

3.
Chinese Journal of Laboratory Medicine ; (12): 340-343, 2016.
Article in Chinese | WPRIM | ID: wpr-494280

ABSTRACT

The early diagnosis of serious infection is very important since the fatality rate is quite high.Traditional diagnostic methods of infection includes procalcitonin,leukocyte count,CRP and so on,of which the sensitivity and specificity is not able to achieve the early diagnosis.Recently,neutrophil CD64 has been widely concerned for the high sensitivity and specificity in early diagnosis of bacterial infection.Flow cytometry is applied to detect neutrophil CD64 in early diagnosis of infectious diseases including respiratory infection,septicemia,neonatal intensive infection,burn and postoperative infection.What's more,the sensitivity and specificity can be further improved if neutrophil CD64 was combined with other inflammatory markers.Thus,neutrophil CD64 detected by flowy cytometry plays an important role in the diagnosis,monitoring,prognosis,therapeutic effect evaluation of infectious diseases.

4.
Medical Principles and Practice. 2012; 21 (1): 68-73
in English | IMEMR | ID: emr-162801

ABSTRACT

The purpose of this study was to clarify the association between P53 and the Bcl-2 family [Bcl-2, Bax, Bcl-xL, Bcl-xS] expression and apoptosis in pancreatic ductal adenocarcinoma [PDAC]. A total of 70 patients with PDAC were studied. The expression of P53 protein in PDAC was assessed using the immunohistochemical method, which categorized the PDAC patients into two groups: group 1: 36 cases with immunonegative P53[-], and group 2: 34 cases with immunopositive P53[+]. The expression of Bcl-2, Bax, Bcl-xL, and Bcl-xS in the 70 PDAC cases was detected by immunohistochemical and Western blotting methods. The apoptotic index [AI] was also measured in these samples by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling [TUNEL] method. The relation between P53 and the Bcl-2 protein family and apoptosis was then evaluated. Bcl-2 and Bcl-xS expression was significantly associated with P53 [p<0.05]. No clear associations were found among P53, Bax and Bcl-xL expression [p>0.05]. The AI of groups 1 and 2 was 12.1 +/- 2.47 and 8.1 +/- 1.48, respectively [p=0.023]. There was no relationship between AI and Bcl-2, Bax, Bcl-xL and Bcl-xS expression [p>0.05, respectively]. Bcl-2/Bax ratio was significantly associated with AI [p<0.01]. Bcl-2 and Bcl-xS represent significant anti-and proapoptotic proteins, respectively, modulated through a P53-dependent pathway in PDAC, and P53 modulated apoptosis mainly through Bcl-2/Bax ratio

5.
Chinese Journal of Hepatobiliary Surgery ; (12): 31-35, 2011.
Article in Chinese | WPRIM | ID: wpr-384782

ABSTRACT

Objective To investigate expression of slug and E-cadherin in pancreatic cancer tissues and determine the inhibitory effects of anti-Slug, an anti-sense plasmid, on the invasion of pancreatic cancer cell lines in vitro. Methods Slug and E-cadherin protein and mRNA was analyzed by IHP and RT-PCR in 36 cases of pancreatic cancer. Then anti-Slug plasmid was transfected into herin and Slug expression. The inhibitory effects of anti-sense Slug were also detected by Transwell motility assay and Matrigel invasion assay. Results The expression of Slug and mRNA in metastatic pancreatic cancer tissue was higher than that in non-metastatic tissue. E-cadherin and mRNA was lower in metastasis tissues(P<0.05). The inverse relationships were further observed by transient transfection of anti-Slug into SW1990H4 cells. The downregulated expression of Slug and re-expression of E-cadherin were found. The Slug mRNA levels were 0.985±0.016,0.973±0.014, 0. 554±0. 011 after 0, 48 h of transfection of anti-sense Slug, and that of E-cadherin were 0.120±0.001, 0.360±0.002, 0. 727±0. 006, respectively. The diference was significant between different time points (P<0.05). The Slug mRNA levels were 0. 206±0.017, 0.968±0.015, and that of E-cadherin were 0. 18±0.002,0.727±0.006 after stable transfection of anti-sense Slug, and control plasmid, respectively. The diference was significant (P<0.05). The motility activity(393±28, 352±24, 96 ±13 )and the invasion activity (223 ± 69, 202 ± 64, 65 ±19) of1 antisense Slug transfectant cells were significantly decreased as compared with those of control cells (P<0.05). Conclusions Higher expression of slug and lower expression of E-cadherin is related to the invasion and metastasis in pancreatic cancer. A reverse corelation of E-cadherin and Slug expression exists in pancreatic cancer. Slug is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human pancreatic cancer.

6.
Chinese Journal of Radiological Medicine and Protection ; (12): 410-413, 2010.
Article in Chinese | WPRIM | ID: wpr-387661

ABSTRACT

Objective To explore the influence of PUMA on radiosensitivity of pancreatic cancer AsPC-1 cells after Slug gene inhibition by transfected short interferencing RNA(siRNA). Methods The AsPC-1 cells were infected with MOI 10,50,100 for 72 h, respectively. The expression of Slug and PUMA was analyzed by Western blotting and immunohistochemistry methods. The transfected and control cells were exposed to 4 Gy γ-rays. The cells inhibition rate was examined by MTT, Hoechst 33342 and IP double staining. DNA ladder and Giemsa staning was used to observe apoptosis. Results The relative value of Slug expression was 0.831 ±0.14,0. 546 ±0.12 and 0.178 ±0.08 after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly lower than that of control group ( F = 4. 992,P < 0.05 ).The relative value of PUMA was 0. 325 ±0. 07,0. 593 ±0. 11 and 0. 978 ±0. 12, after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly higher than that of control group ( F = 4. 324,P < 0. 05 ). The cell proliferation rate was ( 78.76 ± 9. 36 ) % in transfection combined with radiosensitivity group, significantly higher than that of transfection group [ ( 43.68 ± 6.71 ) % ] and radiosensitivity group alone [( 19.25 ± 3.72)% ] (F = 5.056, P < 0.05). The apoptosis of transfection combined with radiosensitivity group was significantly higher than that of others. Conclusions Slug gene targeting siRNA could inhibit the expression of Slug, and consequently increase the activation of PUMA expression, and so enhance the radiosensitivity to γ-rays.

7.
Chinese Journal of Emergency Medicine ; (12): 921-926, 2010.
Article in Chinese | WPRIM | ID: wpr-387037

ABSTRACT

Objective To investigate the expression of nuclear factor-κB (NF-κB) and p53 up-regulated modulator of apoptosis (PUMA) in acute lung injury (ALI) induced by severe acute pancreatitis (SAP), and the therapeutic role of proline dithiocarbamate (PDTC). Method SD rats weighed 200~ 250 g were randomly(random number) divided into sham operation group (A group, n = 18), ALI group (B group, n = 18) and PDTC treatment group (C group, n = 18). The model of SAP was eastablished by injecting 1 mL/kg of sodium tauarocholate into the pancreatic capsule of the rats in B group and C group. The model rats in C group were treated with PDTC one hour after modeling. Six rats of each group were sacrificed 6 h,12 h, and 24 hours after modeling. The histopathological changes in lung and pancreas were observed. The levels of NF-κB p65 and PUMA in lung were detected by using Western blotting, and the expressions of bcl-2, bax and caspase-3 mRNA in the lung were detected by using RT-PCR. The lung tissue was taken for examination under transmission electron microscope. TUNEL was used for detection of apoptotic alveolar epithelial cells. Results Six to 24 hours after modeling, the pathological scores in lung of ALI group were significantly higher than those of control group and PDTC group after sodium taurocholate injection ( P < 0.05). The levels of NF-κB p65 and PUMA, and the expressions of bax and caspase3 mRNA in ALI group at different intervals were higher than those in control group and PDTC group ( P < 0.05),whereas the expression of bcl-2 mRNA in ALI group was lower than that in control group and PDTC group ( P <0.05). The NF-κB p65 was correlated closely and positively with PUMA ( r= 0.987, P < 0.01). Higher activity of caspase-3 acrtive units was seen in ALI group than that in control group and PDTC group ( P < 0.05). The microvilli disappeared in ALI group 24 hours later. The apoptosis index in ALI group was higher than that in control group and PDTC group ( P < 0.05). Conclusions The apoptosis of alveolar epithelial cells of rats in ALI group is caused by PUMA activated by NF-κB. PDTC treatment can inhibit apoptosis of alveolar epithelial cells of rats in ALI group by inhibiting the activation of NF-κB.

8.
Chinese Journal of Microbiology and Immunology ; (12): 65-70, 2009.
Article in Chinese | WPRIM | ID: wpr-381379

ABSTRACT

objective To construct the recombinant adenovirus containing p53 up-regulated modulator of apoptosis(Ad-PUMA)and investigate its growth inhibition effect on pancreatic callCer cells in vitro and in vivo.Methodls Ad-Easy system was used to construct Ad-PUMA by recombination in E.coli.The virus was Dackaged in 293 cells and subsequently identified valid.The AsPC-1 cells were infected with AdPUMA.Before and after Ad-PUMA infection,the expression of PUMA protein wag investigated by western blot,the inhibition rate of AsPC-1 cells was examined by MTY assay.The in vivo tumor suppressive effect was detected in nude mice with human AsPC-1 xenograft.PUMA protein and the apoptosis of AsPC-1 xenograft were detected by western blot and TUNEL(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling)method.Results In vitro,the expression of PUMA protein was increased with titer of Ad-PUMA,the proliferation of AsPC.1 cells were suppressed,significantly,and the effect was in a viral dose-dependent nlanner.In vivo,the growth in nude mice of AsPC-1 infected with Ad-PUMA was significantly inhibited with an inhibition rate of 44.2%.The expression of PUMA was significantly up-regulated,and the apoptosis index wa8 significantiv increased in tumor after Ad-PUMA infection as determined by western blot and TUNEL.Conclusion The expression of PUMA call inhibit the proliferation of pancreatic cancer in vitro and in vivo,and may be used 88 a potential tool for cancer therapy

9.
Chinese Journal of Radiological Medicine and Protection ; (12): 27-30, 2009.
Article in Chinese | WPRIM | ID: wpr-396358

ABSTRACT

Objective To study the effect of PUMA gene mediated by recombinant adenovirus vector combined with radiation on the pancreatic carcinoma. Methods The PANC-1 cells were infected with Ad-PUMA (MOI = 10, 50 and 100, respectively) for 48 h. The expression of PUMA mRNA and protein was detected by RT-PCR and Western blot, respectively. PANC-1 cells were divided into 4 groups: control group, transfection group, irradiation group and combined treatment group. The cell growth inhibition rate and apoptotic rate of PANC-1 cells were assessed by MTT assay and flow cytometry. Human pancreatic carcinomas were transplanted subcutaneously in nude mice, which were randomized into 4 groups: control group, transfection group, irradiation group and combined treatment group. Tumor growth rate and apoptotie index at different time points were recorded in 35 days. Results The expression of PUMA mRNA and protein was increased with the increase of MOI of Ad-PUMA, which was does-dependant (MOI = 10, mRNA = 0.46 ± 0.02, protein = 0. 75 ±0.09;MOI=50, mRNA= 1.12±0.09, protein = 1.01 ±0.18;MOI= 100, mRNA= 1.50±0.08, protein=1.80 ± 0.15 ;P < 0.05). The proliferation of PANC-1 cells was suppressed significantly when transfected by Ad-PUMA in a dose-dependent manner(r = -0.986 55), which was more significant combined with radiation (r = -0.971 26, P < 0.05). Meanwhile, the apoptotie rate was increased in the same manner [for pre- and post-irradiation,which was (45.4 ± 5.26) % and (73.2 ± 6.62) %, respectively, P < 0.05]. From 7 to 35 d after PUMA gene transfection and radiotherapy, the tumor growth was significantly slower than those of irradiation group, transfection group and control group [35 d after therapy, the volume of tumor was (19.82 ± 6.45)mm3 ,(39.5 ± 9.23)mm3 , (33.6 ±3 10.3)mm3 and (52.0 ± 11.43)mm3 , respectively, P < 0.05]. And the apoptotic index was increased in the same manner (AI = 0.43 ± 0.05, 0.29 ± 0.10, 0.24 ± 0.05 and 0.00 ± 0.00, respectively, P < 0.05). Conclusions Recombinant adenoviral-mediated PUMA gene combined with irradiation could increase the cell-killing effect on pancreatic carcinoma. It is better than that of either one kind of therapy.

10.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 1601-1602, 2008.
Article in Chinese | WPRIM | ID: wpr-398262

ABSTRACT

Objective To explore the diagnesticvalue of the inhalant allergens in patients with allergic disea-ses. Methods The intradermal tests(IT) of 20 types of the inhalant allergens which were supplied by Diagnosis Rea-gent Factory of Beijing Union Hospital were made on 1434 cases with allergic diseases. The samples were divided into two age groups:4~18 year-old(children group,n=318) and 19-80 year-old( adults group,n=1116). Distribution of the inhalant allergens of the two groups was investigated. Results A total of 1434 cases (male :702, female:732) were diagnosed with allergic diseases. There were 8 kinds of allergic diseases. The number of allergic rhinitis was the first place( 1130 cases,78.80% ). The first 9 positive allergens separately were: polyvalent pollen Ⅰ (51.05%),house dust(48.81% ) ,mixed inhalant allergens( 48.61% ), artemisia pollen( 46.86% ), dust mite( 45.19% ) , poly-valent insects(36.82% ) ,spring poLlen Ⅰ(33.99% ) ,ragweed pollen(31.24% ) ,spring pollen Ⅱ (30.68% ). There were 1289 cases(89.89% ) at ]east one type positively on allergens. The 19~40 aged patients were the most in dif-ferent age groups. The difference of positive detection rate of allergens was significant in different quarters ( P<0.01 ). Conclusion The youth and middle aged is easoer to suffer loom allergic diseases. Allergic rhinitis was the most common disease,and polyvalent pollen Ⅰ ,house dust,artemisia pollen,dust mite,ragweed pollen were the main allergens.

11.
Chinese Journal of Bases and Clinics in General Surgery ; (12)2003.
Article in Chinese | WPRIM | ID: wpr-544732

ABSTRACT

Objective To investigate the expression changes of nuclear factor kappa B (NF-?B) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha(I?B-?)vector. Methods After pcDNA3-I?B-? vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-?B and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-I?B-? was transfected into HCC9204, the expression of NF-?B was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-?B is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.

12.
China Oncology ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-544631

ABSTRACT

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

13.
Chinese Journal of General Surgery ; (12)1997.
Article in Chinese | WPRIM | ID: wpr-528112

ABSTRACT

Objective To study the relationship of expression of nuclear factor kappab gene and MMP-2mRNA and with lymph node metastasis of pancreatic cancer.Methods The expression of NF-?B was investigated by Western blot in normal pancreatic(NP) samples of 9 cases and pancreatic cancer(PC) samples of 45 cases(lymph node metastasis 30 cases,non-lymph node metastasis 15 cases).The expression of MMP-2mRNA was detected through semiquantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results Of 45 PC cases,the positive expression rate of p65 and MMP-2mRNA was 66.7%(30/45) and(57.7%)(26/45),respectively.Of 30 cases of lymph node metastasis,the positive expression rate of p65 and MMP-2mRNA was 83.3%(25/30)and 73.3 %(22/30),respectively.Of 15 cases of non-lymph node metastasis,the positive expression rate of p65 and MMP-2mRNA is 33.3%(5/15) and 26.7%(4/15),respectively,and the expression of p65 gene was positively correlated with MMP-2mRNA(expression)(r=0.743,P

14.
Chinese Journal of Cancer Biotherapy ; (6)1996.
Article in Chinese | WPRIM | ID: wpr-595738

ABSTRACT

Objective:To investigate the effect of P53 up-regulate modulator of apoptosis(PUMA) on the apoptosis of pancreatic carcinoma BxPC-3 cells and the possible mechanism.Methods: BxPC-3 cells were infected with recombinant adenovirus containing PUMA gene(Ad-PUMA) at 100 MOI for 0-96 h.Apoptosis of BxPC-3 cells was examined by FCM.Expressions of PUMA,Bcl-2,Bax,Cytochrome C and Caspase-3 proteins in BxPC-3 cells were detected by Western blotting.Bax expression in the cytoplasm and mitochondrion and Bax oligomer expression expression in BxPC-3 cells were determined by Western blotting.Results: Apoptosis rates of BxPC-3 cells were significantly increased with the time of Ad-PUMA infection,and peaked after 48 h.Ad-PUMA infection increased the expressions of PUMA,Cytochrome C and Caspase-3 proteins in BxPC-3 cells,and decreased the expression of Bcl-2 protein.Apoptosis rate of BxPC-3 cells after Ad-PUMA infection was correlated with PUMA expression.Ad-PUMA did not affect the expression of total Bax protein in BxPC-3 cells,but Bax expression in cytoplasm was dramatically decreased after infection,and Bax expression in mitochondrion was markedly increased.Furthermore,Ad-PUMA infection induced Bax oligomerization in BxPC-3 cells.Conclusion: PUMA can promote apoptosis of pancreatic carcinoma cells through mitochondrion pathway.

15.
Chinese Journal of Cancer Biotherapy ; (6)1995.
Article in Chinese | WPRIM | ID: wpr-591544

ABSTRACT

Objective:To investigate whether PUMA gene transfection can increase sensitivity of pancreatic cancer cells (PC) to 5-FU-induced apoptosis. Methods: PUMA-pCEP4 containing full length PUMA cDNA or pCEP4 was transfected into human pancreatic cancer cell line AsPC-1 by lipofectamine transfection, G418 selection was used to select positive cells. AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were separately treated with serial concentrations of 5-FU(0.01-100 ?mol/L). MTT assay was used to determine the cell survival rate in each group and IC50 of 5-FU was calculated. TUNEL,FCM and DNA ladder observation were employed to study cell apoptosis. Western blotting was performed to detect the expression of PUMA protein. Results: The 5-FU IC50 values of AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were (12?1.9)?mol/L,(1.6?0.4)?mol/L and (10.4?1.6) ?mol/L, respectively, with the sensitivity of AsPC-1/PUMA cells increased by 7.5 folds. 5-FU induced cell apoptosis of AsPC-1 cells in a dose-dependent manner, with the apoptosis of AsPC-1/PUMA cells more prominent than those of AsPC-1 and AsPC-1/pCEP4 cells. Low concentration of 5-FU (0.1 ?mol/L) induced few apoptosis of AsPC-1/pCEP4 cells([1.14?0.28]%) and AsPC-1 cells ([0.9?0.23]%), and induced apoptosis in AsPC-1/PUMA cells([6.47?1.42]%). High concentration of 5-FU (1.0 ?mol/L) induced apoptosis in all groups, with that in AsPC-1/PUMA cells([34.54?9.36]%) significantly higher than those in AsPC-1/pCEP4 cells([15.8?5.15]%) and AsPC-1 cells ([12.8?3.74]%, both P

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