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@#[Abstract] Objective: To investigate the miR-423-5p expression in brain glioma tissues and cell lines, and its promotive effect on temozolomide (TMZ) chemoresistance by targeting PDCD5 (programmed cell death protein 5). Methods: Tumor tissues and matched peritumoral tissues were collected from 20 brain glioma patients who were surgically treated in the Department of Neurosurgery, Affiliated Hospital of Beihua University between January 2017 and December 2018. Glioblastoma cell lines (U251, U87, SHG-44) and human normal glial cell line HMC-3 were also used in the study. The relative expression of miR-423-5p and PDCD5 in brain glioma and peritumoral tissues and cell lines was detected by qPCR. The synthesized miR-423-5p mimics and miR-NC were respectively transfected into U251 and U87 cells; meanwhile, TMZ at different concentrations (50, 100, 150 and 200 μmol/L) were also used to treat the cells. Then, the chemoresistance of cells to TMZ were determined. MTT assay and colony formation assay were used to examine the proliferation of U251 and U87 cells, andWestern blotting was used to detect the expression of c-caspase 3, Bcl-2 and PDCD5 proteins in U251 and U87 cells. The targeting relationship between PDCD5 and miR-423-5p was validated through Dual luciferase reporter gene assay. Results: miR-423-5p was highly expressed in glioma tissues and glioma cell lines (all P<0.01). As compared with the miR-NC group, the proliferation and TMZ-chemoresistance of U251 and U87 cells in miR-423-5p mimics group significantly increased (all P<0.01). Dual luciferase reporter gene assay validated that miR-423-5p could bind with PDCD5 3' UTR to suppress the expression of PDCD5. Conclusion: High expression of miR-423-5p enhances the chemoresistance of glioma cells to TMZ, and miR-423-5p may serve as a potential therapeutic target in the treatment of brain glioma.
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OBJECTIVE@#To investigate the effect of recombinant human PDCD5 (rhPDCD5) treatment in a rat model of bovine II collagen (CII)-induced arthritis (CIA) on inflammatory cytokine secretion, proliferation and apoptosis of activated lymphocytes and explore the mechanisms of rhPDCD5-induced immunosuppression on activated lymphocytes.@*METHODS@#Female Wistar rats were randomly divided into normal control group, CIA+ ovalbumin (OVA) group, CIA+ rhTNFR: Fc group, and CIA+rhPDCD5 group. The rats in the latter 3 groups received intraperitoneal injections of OVA (14 mg/kg), rhTNFR: Fc (3.5 mg/kg) or rhPDCD5 (14 mg/kg) from day 2 to day 26 following CII injection. On day 28, the spleens of the rats were harvested for preparing single cell suspensions of splenocytes, which were activated by CII (20μg/mL) or anti-CD3 (1μg/mL)+ anti-CD28 (2μg/mL) for 48 h and 72 h. The production of interferon-γ(IFN-γ) and interleukin-17A (IL-17A) by the activated lymphocytes was determined by ELISA of the culture supernatants. The proliferation and apoptosis of the activated lymphocytes were assessed using [H]-thymidine incorporation assay and flow cytometry, respectively.@*RESULTS@#Compared with those in CIA + OVA group, IFN-γand IL-17A secretions by the activated lymphocytes from rhPDCD5-treated CIA rats significantly decreased. RhPDCD5 treatment of the CIA rats obviously suppressed the proliferation and promoted apoptosis of the lymphocytes activated by CII or by anti-CD3 + anti-CD28.@*CONCLUSIONS@#rhPDCD5 reduces pro-inflammatory cytokine secretion, inhibits the proliferation and promotes activation-induced cell death of activated CD4 lymphocytes to produce immunosuppression in rat models of CIA.
Subject(s)
Animals , Cattle , Female , Humans , Rats , Apoptosis , Apoptosis Regulatory Proteins , Arthritis, Experimental , Cell Proliferation , Cytokines , Lymphocytes , Neoplasm Proteins , Rats, WistarABSTRACT
@# Objective: To explore the role of tumor suppressor gene programmed cell death 5 gene (PDCD5) in the growth and temozolomide (TMZ) sensitivity of brain glioma cells. Methods:Atotal of 116 patients with cerebral glioma admitted to the Department of Neurosurgery, First Clinical Hospital of Jilin University from January 2009 to December 2014 were enrolled in this study. QPCR, WB and immunohistochemistry method were used to detect the mRNAand protein expressions of PDCD5 in glioma cell lines (U87, U251), U87 cell line with stable PDCD5 expression (U87-PDCD5), glioma cells with si-PDCD5 transfection and primary cerebral glioma tissues, respectively. MTT assay was used to detect the effect of over-expression or knockdown of PDCD5 on the growth and TMZ-sensitivity of glioma cells. The subcutaneous tumor-bearing model of glioma cell line U87 was established in nude mice, and then the experimental mice were randomly divided into control group, TMZ group, PDCD5 group and TMZ+exogenous PDCD5 recombinant expression vector group.After 20 days, the animals were sacrificed by cervical dislocation and the tumor tissue was excised to measure the tumor volume and weigh. The expression of PDCD5 in tumor tissues was detected by qPCR and WB methods, and the effects of PDCD5 combined with TMZ on the growth of gliomas were also analyzed. Results: The relative mRNA and protein expressions of PDCD5 in U87 cells were significantly lower than those in U251 cells (both P<0.05), and the mRNA and protein expressions of PDCD5 in high level glioma tissues were significantly lower than those in low level tissues (all P<0.05). The sensitivity of U87-PDCD5 cells and U251 cells to TMZ was higher than that of U87 cells (all P<0.05). The sensitivity of cells to TMZ in U87-PDCD5-siRNA group and U251siRNA group was significantly lower than that of the control group (both P<0.05). The tumor volume and weigh to fnudemicexenografts were compared,and the results showed control group>TMZ group>PDCD 5group>combined group(allP<0.05);however, the mRNA and protein expressions of PDCD5 in the transplanted tumor tissues of each group showed the opposite trend (all P<0.05). Conclusion: PDCD5 over-expression can enhance the chemosensitivity of braingliomato the chemotherapy drug TMZ, while silencing of PDCD5 expression exertsthe opposite effect.The combination of PDCD5 and TMZ can better inhibit the growth of xenografts in nude mice.
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Objective To investigate the therapeutic effect of paclitaxel plus oxaliplatin chemotherapy to the transplanted non-small cell lung cancer of nude mice and the effect to the apoptosis protein expression of PDCD5 and XIAP with mice model.Methods A tumor-bearing mice were randomly divided into blank group, normal saline group, oxaliplatin group, paclitaxel group, paclitaxel plus oxaliplatin group.The gene expression of PDCD5 and XIAP was assayed by real-time quantitative PCR(q-PCR).The apoptosis related PDCD5 and XIAP protein were detected by Western blot.Finally, the tumor weight of each group was measured for statistical analysis.ResultsThe mRNA expression of PDCD5 was highest and the gene expression of XIAP was lowest in paclitaxel plus oxaliplatin group(P<0.01).The expression of PDCD5 protein was highest and the expression of XIAP protein was lowest in paclitaxel plus oxaliplatin group (P<0.01).Finally, compare the tumor weight of each group, paclitaxel plus oxaliplatin group has the least mass(P<0.01).Conclusions Paclitaxel plus oxaliplatin group chemotherapy significantly increases PDCD5 expression and reduce XIAP expression.Meanwhile, paclitaxel plus oxaliplatin chemotherapy can significantly reduce the tumor weight of happened non-small cell lung cancer.
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Objective:To investigate the expression of Survivin and programmed cell death 5(PDCD5)protein in mucoepidermoid carcinoma(MEC).Methods:Survivin and PDCD5 were detected by immunohistochemical staining in 20 cases of normal parotid tis-sue(group A),40 cases of pleomorphic adenoma(group B)and 45 cases of MEC tissues(group C).Results:The positive expres-sion ratio of Survivin in group A,B and C was 10.0%,27.5% and 55.6% respectively(χ2 =14.556,P <0.01),while that of PD-CD5 was 85%,65% and 33.3% respectively (χ2 =17.439,P <0.01).The expression of survivin or PDCD5 was related with dif-ferentiation,lymph node metastasis and TNM staging of MEC.Survivin and PDCD5 showed a negative correlation in MEC(χ2 =4.500,P =0.034,γs =-0.316).Conclusion:Survivin over-expression and PDCD5 down-expression may play a role in the de-velopment and progress of mucoepidermoid carcinoma.
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Programmed cell death 5 ( PDCD5 ),formerly designated as TFAR19 ( TF-1 cell apoptosis-related gene 19),is an apoptosis-regulated gene cloned by Peking University Center for Human Disease Genomics.PDCD5 is expressed in many human tissues,with a high degree of homology,plays a regulatory role during cell apoptosis.Disorders of PDCD5 expression is correlated with tumorigenesis.This article reviews the structure,expression and functions of PDCD5,makes a summary of its relationship with kinds of tumors.Further studies about clinical application of PDCD5 are needed.
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Programmed cell death 5(PDCD5)is a novel gene which can promote and regulate apoptosis. The evidences of some studies show that PDCD5 displays reduced expression in a variety of tumor tissue. Increased expression of PDCD5 can improve the sensitivity of anti-tumor chemotherapy. Because of this feature with PDCD5,it is expected to become a new chemosensitizer.
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Objective To observe apoptosis and its relationship between the expression of caspase-3 and programmed cell death 5 (PDCD5) protein in perihematoma tissue after intracerebral hemorrhage in rats and to investigate the injury mechanism after intracerebral hemorrhage. Methods A total of 54 male Wistar rats were randomly divided into sham operation and intracerebral hemorrhage groups, and the latter were redivided into 3,6, 12 hours, days 1,2,3, 5, and 7 subgroups (n = 6 in each group). A model of intracerebral hemorrhage was induced by injecting 50 μL autologous tail artery blood into the caudate nucleus. Apoptosis was detected by TUNEL method. The expressions of Caspase-3 and PDCD5 was observed by immunohistochemistry. Results The apoptotic cells were found in perihematoma tissue of rats at 3 hours,they reached the peak at day 2 to day 3 and reduced gradually after 3 days. Caspose-3 and PDCD5 positive cells were found in perihematoma tissue of rats at 3 hours, they reached peak at day 1 to day 2 and reduced gradually after 3 days. The numbers of Caspase-3 (r =0. 971, P ＜0. 01 ) and PDCD5 (r = -0. 334, P ＜0. 01 ) positive cells were positively correlated with those of apoptotic cells in perihematoma tissue after intracerebral hemorrhage in rats. Conclusions The perihematoma tissue of intracerebral hemorrhage in rats existed apoptosis, and it was consistent with the expressions of Caspase-3 and PDCD5. Caspase-3 and PDCD5 may promote apoptosis in perihematoma tissue after intracerebral hemorrhage.
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Objective: To study the expression of PDCD5 mRNA and its significance in esophageal cancer in Xinjiang Kazakh and Han nationality. Methods: RT-PCR was used to detect the mRNA of PDCD5 in 40 cases of esophageal cancer (18 cases of Kazakh, 22 cases of Han). Results: The positive rate of PDCD5 mRNA expression in 40 samples of esopha-geal cancer tissue, adjacent tissue, and normal tissue was 80.0% (32/40), 80.0% (32/40) and 87.5% (35/40), respectively, with no significant difference (P>0.05). The Ods of cancer tissues, adjacent tissues, and normal tissues were 0.7644± 0.1444, 0.9341 ±0.1631 and 1.8703±0.4767, respectively. The expression of PDCD5 was significantly increased in cancer tissues compared with that in normal tissues (P<0.05). The expression level of PDCD5 mRNA was not significantly correlat-ed with the degree of differentiation and lymph node metastasis. Conclusion: No significant difference was found in PDCD5 mRNA expression in esophageal cancer between Xinjiang kazakh and Han nationality (P>0.05). The expression of PDCD5 is not correlated with the degree of differentiation, depth of invasion or lymph node metastasis. Detection of PDCD5 mRNA expression in esophageal cancer tissues may provide valuable information for patient prognosis.
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Objective To observe the effect of allogenic bone marrow mononuclear cells(BM-MNCs) transplantation on myocardial apoptosis after acute myocardial infarction(AMI) in rats.Methods 40 Wistar rats were randomly divided into control group(n=20) and transplantation group(n=20).Myocardium around the infarcted left ventricular area of the rats in transplantation group were injected with BM-MNCs suspension beneath the epicardium.Myocardium the area of control group was injected with culture solution.Results After 4 weeks of the operation,the myocardial apoptosis index,the TNF-? content and the PDCD5 mRNA of transplantation group were all notably less than those of control group(P
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Objective: To investigate the expression of PDCD5 in tissues of normal cervix, CIN Ⅰ-Ⅲ,cervical cancer and explore the relationship between PDCD5 and cervical cancer.Methods: After we defined the most fitful condition, tissues from 18 cases of normal cervix, 19 of CIN Ⅰ, 18 of CIN Ⅱ, 20 of CIN Ⅲ and 18 of cervical cancer were defined by indirect immunohistochemical technique. Positive expression rates and intensity of PDCD5 protein were investigated by observing under microscope and analyzing with computer imaging technique. The results were analyzed with one way anova. Results: The results of immunohistochemical staining showed that the percentage of strong positive cells in normal cervical tissue and CIN Ⅰ were significant higher than those of CIN Ⅱ, CIN Ⅲ and cervical cancer. On the whole of the condition of immunohistochemical staining, the expressions of PDCD5 were downregulated along the progression of cervical atypical epithelia, but that in CIN Ⅰ was upregulated.The ODs of normal cervix,CIN Ⅰ-Ⅲ,cervical cancer were 0.322,0.366,0.287,0.252,and 0.206 respectively.The intensity of each group showed obvious differences.Conclusion: We found that the expression of PDCD5 was upregulated in CIN Ⅰ and downregulated in CIN Ⅱ, CIN Ⅲ and cervical cancer. It suggests that PDCD5 is an important apoptosis regulating factor in the occurrence of cervical cancer.
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Objective:To explore the effect of progrmmed cell death 5(PDCD5)on apoptosis of rheumatoid arthritis fibroblast-like synoviocytes(RA FLS) induced by triptolide.Methods:Cultured synovial cells in vitro from RA patients were transfected with Ad-PDCD5.At protein level,expression of PDCD5 in RA FLS infected with Ad-PDCD5 was detected by Western blot.RA FLS infected with Ad-PDCD5 were cultured in presence or absence of triptolide and apoptosis of RA FLS was determined by flow cytometry.Results:Infection of RA FLS with increasing concentrations of Ad-PDCD5(50-300 MOI) resulted in a does-dependent increase in the production of PDCD5.Apoptotic cells percentage for noinfection group,Ad-null group and Ad-PDCD5 group were(22.41?3.87)%,(28.77?12.97)% and(48.87?12.69)%,respectively.Alternatively,infection without addition of triptolide stimuli had no effect.The data showed that gene transfection of PDCD5 alone without addition of triptolide was not sufficient to activate RA FLS apoptosis,PDCD5 acted as an enhancer rather than inductor of apoptosis.Conclusion:Overexpression of PDCD5 could enhance apoptosis of RA FLS induced by triptolide,PDCD5 may be a potential therapeutic target to RA.
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Objective:To lay foundation for the functional studies of programmed cell death 5 ( PDCD5 ) and develop new technical pathway for bioinformatics analysis of human functional genes. Methods:Using PDCD5 as the target molecule, intensive bioinformatics analysis of the nucleic acid and protein sequences were conducted. Data mining and comprehensive analysis by sequence against database similarity searching, ortholog structure comparison, expression profile analysis and gene “neighbor” listing were performed. Results: Two human putative pseudogenes on chromosomes 12 and 5, and one mouse putative pseudogene on chromosome 1 were identified. The methanobacterium thermoautotrophicum ortholog was classified as the same fold as ubiquitin and ribosomal protein S13. The C. elegans ortholog, ubiquitin and IAP (inhibitor of apoptosis proteins) belonged to the same expression profile cluster. This cluster was related to biosynthesis and protein synthesis. PDCD5 orthologs in various genomes were adjacent to various ribosomal proteins on the chromosome. Conclusion: The human genome contains at least two processed pseudogenes of PDCD5 . Besides the relationship with cell apoptosis, PDCD5 is predicted to have functional relationship with ubiquitin and participate in the translation regulation.
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Objective:To set up an effective and simple purification method to obtain highly purified prokaryotic protein of PDCD5 and study its stability. Methods:Recombinant PDCD5 protein expressed in E. coli was accumulated as an inclusion body. After washing, the inclusion body was denatured, renatured, digested with thrombine and then purified by two steps of chromatography. The purity of the products was analyzed by capillary electrophoresis and the stability was identified by SDS-PAGE. Results:Capillary electrophoresis showed that the purity of protein was 100%, and molecular weight was 15 800 with pI 5.9. Further bioactivity assay indicated that the purified PDCD5 could enhance the apoptosis of HL 60 cells withdrawing cytokine, which was in a dose dependent manner. Stability analysis showed that the PDCD5 protein was sensitive to temperature and easy to degrade at 4 ℃ and 25 ℃. However, it was relatively stable at -20 ℃ or lyophilized. Conclusion:Highly purified and stable recombinant PDCD5 protein was obtained, which lays a foundation for the functional study and application investigation of PDCD5 .