Role of inhibiting lncRNA TUG1 to down⁃regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease / 解剖学报

Objective To investigate the relieving effects of knockdown of long non-coding RNA(lncRNA)taurine up-regulated gene 1 (TUG1) on inhibiting nucleotide binding oligomerization domain like receptor protein 1 (NLRP1) inflammasome and the progression of Alzheimer’ s disease. Methods Wild-type (WT group, 10 mice) or amyloid precursor protein (APP) / presenilin-1 (PS1) transgenic mice (30 mice) with a genetic background of C57 / BL6 aged 9-10 weeks were used in this study. APP / PS1 transgenic mice were randomly divided into model group, model+lncRNA TUG1 short hairpin RNA (shRNA) group and model + shRNA non target (NT) group (n = 10) . Blood samples, cerebral cortex tissues, primary microglial cells and primary astrocytes were collected from mice 12 weeks of age on day 1 (3-month-old) and 32 weeks of age on day 1 (8-month-old), with 5 mice per group at each time point. Real-time PCR analysis was used to detect the expression levels of lncRNA TUG1 and macrophage migration inhibitory factor (MIF) mRNA in cerebral cortex tissues and primary microglial cells, and C1r and C1s mRNA levels in primary astrocytes of 3-month-old and 8-month-old mice in the above 4 groups, respectively. ELISA was used to determine the MIF in plasma samples of the above 4 groups of mice. Primary microglia and astrocytes from the cerebral cortex of 3-month-old and 8-month-old mice were co-cultured. CCK-8 method was used to determine the proliferation ability of the above cells. Western blotting was used to determine the expression levels of MIF, pro interleukin-1β (pro-IL-1β), apoptosis associated speck-like protein containing a caspase recrult domain(ASC), Caspase-1 (p20), Caspase-1 (full), NLRP1 and NLRP3 in cerebral cortex tissues of 3-month-old and 8-month-old mice. Immunofluorescent staining was used to determine amyloid beta(Aβ) in cerebral cortex of 8-month-old mice. Results At the age of 3-month-old and 8-month-old, compared with the WT group, the relative expression level of lncRNA TUG1 and MIF in cerebral cortex tissues and primary microglia of model group mice was significantly up-regulated, with primary microglial cells and astrocytes proliferation ability enhanced (P0. 05) . There was no significant difference between the model group and the model+shRNA NT group mice of all the above factors (P>0. 05) . Conclusion In APP / PS1 transgenic mice, up-regulation of lncRNA TUG1 and MIF are positively associated with the activation of NLRP1 inflammasome in mice cerebral cortex tissues and primary microglia. Knock-down of lncRNA TUG1 can ameliorate the progression of Alzheimer’ s disease.

Screening chronic periodontitis-related genes using whole-genome expression profiling / 中国组织工程研究

BACKGROUND: Whole-genome expression profiling is a technical method for gene expression research, with high sensitivity and specificity. This technique can be used to detect differential genes related to chronic periodontitis in the whole genome, therefore efficiently and quickly finding chronic periodontitis-related factors. OBJECTIVE: To screen genes related to chronic periodontitis by using the whole-genome expression profiling. METHODS: Normal periodontal ligament tissue of 15 patients with orthodontic extraction was selected as control group, and periodontal tissue of 21 patients with chronic periodontitis was selected as experimental group. To screen up-regulated and down-regulated genes. the genome-wide expression profile chips of four chronic periodontitis tissues and four healthy tissues were compared. The expression of the differential gene PI3K-Akt signal pathway was verified by real-time PCR (7 normal cases and 13 cases of chronic periodontitis) and western blot (4 normal cases and 4 cases of chronic periodontitis). The experimental protocol was approved by the Ethics Committee of the First Affiliated Hospital of Hainan Medical University (approval No. HNM20180034) and informed consent was obtained from each patient. RESULTS AND CONCLUSION: Analysis of the whole genome expression profile chip revealed that 1 565 up-regulated genes and 1 849 down-regulated genes were significantly differentially expressed in chronic periodontitis samples. The enrichment analysis revealed that the expression of PI3K-Akt signaling pathway was significantly different in chronic periodontitis (P < 0.001). Real-time PCR and western blot assay results indicated that PI3K and Akt expression was higher in the experimental group than in the control group (P < 0.05). All the findings indicate that the genome-wide expression profile chip is fast and highly sensitive to screen the changes in chronic periodontitis-related genes. Significantly differential expression of PI3K-Akt signal pathway in chronic periodontitis provides an experimental basis for the treatment of chronic periodontitis.

Effects of LncRNA HULC on radiosensitivity of osteosarcoma cells / 中华放射医学与防护杂志

Objective To investigate the effect of LncRNA HULC on radiosensitivity of osteosarcoma cells. Methods Osteosarcoma cells OS732 was infected by shRNA HULC lentivirus, and the interference effect was determined by qRT-PCR. Osteosarcoma cells infected with shRNA HULC lentivirus were irradiated with 8 Gy X-rays. MTT, PI monochrome staining and Annexin V-FITC/PI double staining were used to detect cell proliferation, cell cycle and apoptosis, respectively. Western blot was used to detect the protein levels of p21, Cyclin D1, C-Caspase-3 and Cyt-C in cytoplasm and mitochondria. Plate cloning assay was used to evaluate cell radiosensitivity. Results The expression of HULC in osteosarcoma cells was significantly down-regulated by shRNA HULC lentivirus infection. Down-regulation of HULC or irradiation inhibited osteosarcoma cell proliferation [(100. 00±9. 65)％ vs. (71. 36±5. 27)％, (63. 48± 5. 93)％, t=4. 512, 5. 585, P<0. 05 ] , blocked cell cycle [ ( 50. 15 ± 5. 14 )％ vs. ( 62. 35 ± 4. 22 )％, (66. 05±5. 23)％,t=3. 177,3. 756,P<0. 05], induced cell apoptosis [(2. 98±0. 23)％ vs. (22. 61± 3. 26)％, (26. 14±2. 81)％,t=8. 898,10. 498,P<0. 05], promoted the expressions of p21 and Cyclin D1 in cells, down-regulated the level of C-Caspase-3 protein, increased the level of Cyt-C protein in cytoplasm, and down-regulated the level of Cyt-C protein in mitochondria. Downregulation of HULC combined with irradiation yield much more effects on cell proliferation inhibition [ ( 71. 36 ± 5. 27 )％, (63.48±5.93)％ vs. (49.32±5.76)％, t=4.890, 2.967, P<0.05], cell cycle arrest [(62.35± 4. 22)％, (66. 05±5. 23)％ vs. (77. 17±7. 54)％, t=2. 983, 2. 106, P<0. 05], apoptosis induction [(22. 61±3. 26)％, (26. 14±2. 81)％ vs. (36. 21±3. 26) ％,t=6. 164, 4. 564, P<0. 05] and the expressions of p21, Cyclin D1, C-Caspase-3 and Cyt-C in osteosarcoma cells. The radiosensitization ratio of down-regulation of HULC was 1. 432. Conclusions Down-regulation of HULC enhances radiosensitivity of osteosarcoma cells, which may be related to cell cycle arrest and apoptosis induction.

Bioinformatics Analysis of Up-regulated Differential Genes in Rats post Spinal Cord Injury / 中国康复理论与实践

Objective:To investigate the differential expression and gene functions of up-regulated genes in rats with spinal cord injury. Methods:Female Sprague-Dawley rats' model of spinal cord injury was established with the modified Allen's method. Gene chip technology was used to detect the variation of differentially expressed genes in the spinal cord after spinal cord injury in rats. The differences in genes, functional localization and pathways were analyzed with gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway database. Results:The results of total RNA quality in spinal cord segment were qualified. Gene chip results showed that there were 1874 differentially up-regulated genes and 2348 differentially down-regulated genes. Bioinformatics was used to analyze differentially up-regulated genes in terms of biological processes, cellular components, and molecular functions. The differentially up-regulated genes were involving apoptosis, immune response, inflammation, etc., pathway analysis mainly showed the differentially up-regulated genes involved phosphoinositide 3-kinase protein kinase B signaling pathway and Toll-like receptor signaling pathways. Conclusion:Differentially up-regulated genes may be involved in secondary reactions following spinal cord injury, such as inflammation, immune response and hypoxia, and then further affect motor function and sensory function.

Effect of lncRNA HULC knockdown on rat secreting pituitary adenoma GH3 cells

Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.

Effect of Jiawei-Naotai formula on ATF4/CHOP/Puma pathway of ovariectomized rats with cerebral ischemia / 中国病理生理杂志

AIM:To investigate the effects of Jiawei-Naotai formula (JWNTF) on ATF4/CHOP/Puma pathway in hippocampal neurons of ovariectomized female rats with cerebral ischemia.METHODS:The female rats were randomly divided into sham group, model group, JWNTF group and positive control group.The rats, expect in the sham group, were ovariectomized.The rats in each group were intragastric administration 11 days after ovariectomy.The rats in sham group and model group were given a gavage of 0.9％Na Cl, while the rats in other groups were administrated by corresponding therapy intragastrically for 3 d.The regional cerebral ischemia model was established by middle cerebral artery occlusion (MCAO) suture method 14 days after ovariectomy.The behaviors of the rats were evaluated 24 h after cerebral ischemia.The mRNA levels of Bax, Bcl-2 and caspase-3 were detected by RT-qPCR, and the protein expression of Bax, Bcl-2, caspase-3, ATF4, CHOP and Puma was determined by Western blot.RESULTS:Compared with sham group, the neurobehavioral scores significantly increased in other groups (P<0.05).Compared with model group, the neurobehavioral scores were significantly decreased in positive control group and JWNTF group (P<0.05).The protein expression of Bax, caspase-3, ATF4, CHOP and Puma, and the mRNA expression of Bax and caspase-3 in the hippocampus were much higher, and Bcl-2 was lower in model group than those in sham group (P<0.05).JWNTF significantly reduced the protein expression of Bax, caspase-3, ATF4 and CHOP, and the mRNA expression of Puma, Bax and caspase-3, and markedly increased the expression of Bcl-2 at mRNA and protein levels compared with model group.CONCLUSION:The JWNTF protects against brain damage induced by cerebral ischemia, which may be related to inhibitiing the expression of ATF4/CHOP/Puma pathway-related molecules at mRNA and protein levels.

Effects of long non-coding RNA TUG1 on the proliferation and apoptosis of gastric cancerAGS cells / 中国肿瘤生物治疗杂志

@#Objective: To investigate the expression of lncRNA TUG1 (long non-coding RNA taurine up-regulated gene 1) in gastric cancer and its effect on the proliferation and apoptosis of gastric cancer cells. Methods: Surgically resected gastric cancer tissues and corresponding distal normal tissues (>5 cm away from the margin of tumor) of 40 gastric cancer patients from March 2016 to December 2017 at Ganzhou People's Hospital of Jiangxi Province were collected, and qPCR was used to examine the expression of lncRNA TUG1.AGS gastric cancer cells were transfected with lncRNATUG1 over-expression plasmids and siRNAs, and the effects of lncRNA TUG1 on cell proliferation and apoptosis were assessed by CCK-8, qPCR and Flow cytometry. Results: lncRNATUG1 expression was significantly increased in gastric cancer tissues as compared to normal tissues; and it was not correlated with gender, age, tumor size, infiltration depth of tumor, lymph node-metastasis, tumor differentiation and TNM staging. TUG1 over-expression significantly suppressed the expressions of CDKN1A, BAX and Caspase-3 in AGS gastric cancer cells, and decreased G1 phase proportion and apoptosis rate, but increased S phase proportion and cell viability; in contrast, TUG1 siRNA transfection significantly promoted the expressions of CDKN1A, BAX and Caspase-3, and increased G1 phase proportion and apoptosis rate, but decreased S phase proportion and cell viability. Conclusion: Up-regulated lncRNATUG1 promotes proliferation and inhibits apoptosis of gastric cancer cells.

Bioinformatics Analysis of long non-coding RNA TUG1 in hepatocellular carcinoma / 国际检验医学杂志

Objective To explore the significance of long non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) in hepatocellular carcinoma (HCC),to predict the target gene of TUG1,and to provide a ref-erence for further study of TUG1 in HCC.Methods The differential expression of TUG1 in HCC was ana-lyzed by using the UALCAN database and the survival analysis of TUG1 was performed.The target gene of TUG1 was predicted by RegRNA 2.0 biology software,HMDD,targetscan and microT-CDS,and the regulato-ry network of lncRNA TUG1-microRNAs-mRNAs was constructed.The predicted target gene was analyzed by Gene Ontology (GO) and KEGG signal transduction pathway enrichment by using FunRich platform. Results TUG1 expression in HCC was significantly increased,and the expression level of TUG1 increased generally with the increase of tumor grade.The overall survival of patients with low expression of lncRNA TUG1 was significantly longer than that of lncRNA TUG1 high expression patients.There were four possible binding sites of HCC related microRNAs (hsa-mir-122-5p,hsa-mir-200a-3p,hsa-mir-34c-3p,hsa-mir-629-3p) on TUG1,which regulated 245 downstream target genes and formed the regulatory network of lncRNA TUG1-microRNAs-mRNAs.In the biological process,microRNA target genes were highly enriched in the processes such as the regulation of nucleobase,nucleoside,nucleotide and nucleic acid metabolism.In KEGG pathway analysis,microRNA target genes were highly enriched to the signal pathways mediated by Syndecan and TRAIL.Conclusion TUG1 expression level in HCC increased.Increased expression of TUG1 is associat-ed with poor prognosis in HCC.Bioinformatics methods can be used to explore the mechanism of tumorigene-sis from the molecular level,which can provide valuable information for subsequent experiments and clinical diagnosis and treatment.

Silencing pancreatic adenocarcinoma up-regulated factor increases the sensitivity of pancreatic cancer cell line to gemcitabine / 中华肝胆外科杂志

Objective To observe the influence on the sensitivity of pancreatic cancer cell line BxPC-3 to gemcitabine of silencing PAUF gene.Methods BxPC-3 cells,which overexpress PAUF,was stably transfected with PAUF-shCtrl and PAUF-shRNA to establish BxPC-3_shCtrl and BxPC-3_shPAUF cells as control and experiment group.Then the mRNA and protein expression level of PAUF in these two cell lines were detected by RT-PCR and western blot,respectively.The growth inhibition rates of these two cell lines treated with different concentrations of gemcitabine (0,3.1,6.25,12.5,25,50,100,200 nmol/L) were detected by MTT.Apoptosis rates in the cells treated with different concentrations of gemcitabine (0,75,100 nmol/L) were then observed by flow cytometry.Results The relative PAUF mRNA expression level in BxPC-3_shCtrl and BxPC-3 cells were 1.00 ± 0.06 and 0.83 ± 0.07,which were significantly high er than that in BxPC-3_shPAUF cells (0.25 ± 0.02;both P ＜ 0.05).The relative PAUF protein expression level in BxPC-3_shCtrl and BxPC-3 cells were 0.89 ± 0.07 and 0.95 ± 0.04,which were significantly high er than that in BxPC-3_shPAUF cells (0.31 ± 0.03;both P ＜ 0.05).The IC50 value of gemcitabine to BxPC-3_shCtrl cell was (22.88 ± 2.43) nmol/L,which was significantly higher than that of BxPC-3_shPAUF cells [(1.06 ± 0.02) nmol/L;P ＜ 0.05];apoptosis rate of BxPC-3_shPAUF cells treated by gemcitabine increased faster than that of BxPC-3_shCtrl cells.Conclusion PAUF silencing could greatly enhance the sensitivity of BxPC-3 cells to gemcitabine.

Regulatory effects of taurine up-regulated gene 1 on tumorigenesis / 中国病理生理杂志

It has been estimated that approximately 75% of the human genome is transcribed into RNA, 74% of which would be transcribed into non-coding RNA (ncRNA).The ncRNA can be divided into 2 major groups including small RNA and long non-coding RNA (lncRNA).There is increasing evidence that the dysregulation of lncRNA is closely associated with the occurrence and progression of many tumors.The lncRNA taurine up-regulated gene 1 (TUG1) is originally detected in a genomic screen for genes in response to taurine treatment of developing mouse retinal cells.According to research reports, dysregulation of TUG1 participates in the progression of a variety of tumors.Therefore, the regulatory effects of lncRNA TUG1 on tumorigenesis are summarized in this article.

Effects of VDUP1 on proliferation and migration of human breast cancer MCF-7 cells / 中国病理生理杂志

AIM:To investigate the effect of vitamin D3 up-regulated protein 1 (VDUP1) gene over-expression/knockdown on the proliferation and migration of human breast cancer MCF-7 cells and its related mechanisms.METHODS:Gene over-expression/interference techniques were used to up-regulate/down-regulate the expression of VDUP1 in the MCF-7 cells.The mRNA expression of VDUP1 was detected by qPCR.CCK-8, BrdU and Transwell assays were used to measure the cell viability, proliferation and migration, respectively.The protein levels of Akt, p-Akt, GSK3β and p-GSK3β were determined by Western blot.RESULTS:The mRNA expression of VDUP1 was up-regulated after transfection with VDUP1 over-expression plasmid (P<0.05), and down-regulated after transfection with VDUP1 siRNA (P<0.05).Over-expression of VDUP1 significantly inhibited MCF-7 cell proliferation and migration (P<0.05), while knockdown of VDUP1 enhanced cell proliferation and migration (P<0.05).Furthermore, over-expression of VDUP1 up-regulated the protein levels of p-Akt and p-GSK3β (P<0.05).Inverse results were obtained after knockdown of VDUP1.CONCLUSION:The viability and migration ability of MCF-7 cells are inhibited by over-expression of VDUP1 but enhanced by VDUP1 knockdown, which may be related with Akt/GSK3β pathway.

Effect of URG11 gene silencing on proliferation and invasion abilities of human prostate cancer cells / 中国病理生理杂志

AIM:To investigate the expression of up-regulated gene 11 ( URG11 ) in prostate cancer cell line and the effect of URG11 siNRA on the proliferation and invasion of human prostate cancer LNCaP cells.METHODS:The mRNA and protein levels of URG11 in prostate cancer cell lines and normal prostate epithelial cell line were evaluated by real-time PCR and Western blot.LNCaP cells were transfected with designed siRNA using the liposome method.The prolif-eration, apoptosis, migration and invasion abilities of the LNCaP cells were evaluated by MTS assay, flow cytometry, wound-healing assay and Transwell assay.RESULTS:The expression of URG11 at mRNA and protein levels in the DU145, PC3, LNCaP cell lines was significantly higher than that in RWPE-1 cell line.Compared with the control group, the proliferation of LNCaP cells with URG11 siRNA was stagnant in G1/S phase and induced apoptosis.The proliferation of LNCaP cells at 0 h, 24 h, 48 h and 72 h was inhibited after URG11 expression was down-regulated (P<0.05).Transwell assay showed that migration (P<0.05) and invasion (P<0.05) were also inhibited.CONCLUSION:URG11 is highly expressed in prostate cancer.Silencing of URG11 significantly inhibits the proliferation and invasion and induces apoptosis of LNCaP cells.

Expression of taurine up-regulated gene 1 and the clinical significance in renal cell carcinoma / 中国综合临床

Objective To detect the expression level of Taurine up?regulated gene 1( TUG1) in the re?nal cell carcinoma and paired paracancerous normal tissues,then explore the relationships between the expression level of TUG1 and clinical characteristics.Methods RNA was Extacted from the resected renal cell carcinoma tissues and paired paracancerous normal tissues of 46 patients respectively,by reverse transcription to get cDNA, the expression level of the TUG1 was detected by RT?qPCR, the relationship between the expression level of TUG1 and the clinicopathological characteristics was analyzed by statistically software. Results The expression of TUG1 in renal cell carcinoma was obviously lower than that in paired paracancerous normal tissues(0.533±0. 027 vs. 1.000±0.298,t=-3.350,P0.05).Conclusion The expression of TUG1 in renal cell carcinoma tissues are down?regulated,which also suggest that it may be re?lated to the tumorigenesis and development of renal cell carcinoma.

Expression of AEG-1-1 gene in NSCLC and its clinical significance / 中国免疫学杂志

Objective:To investigate the expression of AEG-1 gene in NSCLC and its clinical significance. Methods:Selected our hospital cardiothoracic surgical resection of 83 cases of postoperative cancer tissues of NSCLC patients and 20 paracancer to study, immunohistochemical staining was used to detect the expression level of AEG-1 protein in two groups,the clinical and pathological of AEG-1 protein in patients with NSCLC was analyzed. Results:NSCLC tissues AEG-1 protein expression 46 cases ( 55. 42%) was sig-nificantly higher than 2 cases ( 10. 00%) of paracancer ( P0. 05 ) . AEG-1 high expression of NSCLC in patients with a median survival time of 15. 0 months was significantly lower than that of 19. 0 months (log-rankχ2=4. 119 P<0. 05,) in patients with low expression of AEG-1. Conclusion:AEG-1 gene expression has been up-regulated in NSCLC tissue,which was related to the clinical stage and distant metastasis of the patients.

Mechamism of PAUF in pancreatic cancer pathogenesis and progression / 中华肝胆外科杂志

Pancreatic adenocarcinoma up-regulated factor(PAUF),a newly discovered gene,is highly expressed in pancreatic cancer.PAUF promotes the metastasis and progression of pancreatic cancer through many ways,such as the activation of signal pathway (CXCR4,β-catenin,TPL2/MEK/ERK,FAK/Scr),increasing the adhesiveness of pancreatic cancer cells,promoting angiogenesis and vascular permeability.Simultaneously,CXCR4,β-catenin,TPL2/MEK/ERK and FAK/Scr are closely related with gemcitabine-resistance.Based on this theory,we infer that PAUF plays a role in gemcitabine-resistance of pancreatic cancer cells.So far,no related research has been done domestic and overseas.The research may find a clue for the mechanism of chemotherapy-resistance and provide a new target spot for the therapy of pancreatic cancer.

BH3-only members are involved in radiation-induced apoptosis in mouse liver / 第二军医大学学报

Objective To study the expression of apoptosis-associated genes and the role of apoptosis in acute radiation-induced liver injury. Methods A mouse radiation model, which was irradiated with 60Co γ ray, was established in this study, and the pathological changes were observed by light and electron microscopy for 48 hours. Western blotting analysis was used to measure the expression of some proteins (Caspase 3,Caspase 8, Bcl-2, Bcl-xL, Bax, Bad, PUMA, Slug) and TUNEL assay was employed to examine cell apoptosis in mouse liver. Results Degeneration and apoptosis were found in the liver at 4 h after irradiation and necrosis occurred at 12 h after irradiation. The peak of apoptosis with activation of Caspase 3 in liver was detected during 24-48 h after irradiation. Bcl-2 protein expression was up-regulated during 4-24 h after irradiation, then was down-regulated at 48 h; PUMA protein was up-regulated during 4-48 h after irradiation; Bcl-xL and Bad protein were up-regulated during 6-48 h after irradiation; and Bax and Slug protein were up-regulated only at 12 h after irradiation. Conclusion Up-regulation of BH3-only members (PUMA and Bad) after irradiation may be associated with the increase of apoptotic cells in the liver.

Vitamin D3 up-regulated protein 1 controls the priming phase of liver regeneration

Vitamin D3 up-regulated protein 1 (VDUP1) is a potent growth suppressor that inhibits tumor cell proliferation and cell cycle progression when overexpressed. In a previous study, we showed that VDUP1 knockout (KO) mice exhibited accelerated liver regeneration because such animals could effectively control the expression of cell cycle regulators that drive the G1-to-S phase progression. In the present study, we further investigated the role played by VDUP1 in initial priming of liver regeneration. To accomplish this, VDUP1 KO and wild-type (WT) mice were subjected to 70% partial hepatectomy (PH) and sacrificed at different times after surgery. The hepatic levels of TNF-alpha and IL-6 increased after PH, but there were no significant differences between VDUP1 KO and WT mice. Nuclear factor-kappaB (NF-kappaB), c-Jun-N-terminal kinase (JNK), and signal transducer and activator of transcription 3 (STAT-3) were activated much earlier and to a greater extent in VDUP1 KO mice after PH. A single injection of TNF-alpha or IL-6 caused rapid activation of JNK and STAT-3 expression in both mice, but the responses were stronger and more sustained in VDUP1 KO mice. In conclusion, our findings provide evidence that VDUP1 plays a role in initiation of liver regeneration.

PUMA enhanced chemotherapeutic sensitivity of human glioma stem cells against temozolomide / 肿瘤研究与临床

Objective To explore differences of drug resistance of temozolomide(TMZ) on different CD133 immune prototype of glioma cells and study on the changes of their sensitivity to TMZ through increased PUMA. Methods CD133+-U87MG cells sorted by CD133 magnetic beads were cultured in serum-free stem cell medium respectively. The cells were infected with recombinant adenovirus, Ad-PUMA, diluted in cell culture medium with or without TMZ intervention.The inhibitory rate of cell proliferation was detected by MTT assay and 50 ％ inhibition concentration of TMZ was calculated. Apoptosis rates of CD133+-U87MG cells were assessed by flow cytometry (FCM) before and after intervention of exogenous PUMA and TMZ.Results The TMZ IC50 values of CD133+glioma cells were higher than that of CD133- glioma cells. There were significant differences in apoptosis rate between CD133+ glioma cell and CD133- glioma cell (all P＜0.05).Conclusion AdPUMA joint TMZ can promote glioma stem cells apoptosis, thus improve the sensitivity to chemotherapy of glioma.

Study on effect of recombinant adenovirus PUMA on sensitivity of human glioblastoma cells to temozolomide in vivo / 肿瘤研究与临床

Objective To investigate the inhibitive effects of Ad-PUMA combined with temozolomide on human glioblastoma cells growth in vivo experiments. Methods The nude mouse model with human glioblastoma cells subcutaneous transplantation was established. The mice were randomly divided into 4 groups to receive subcutaneous injection at the 14th day separately with: Normal saline 100 μl (control, n=8), Ad-PUMA 2×108 pfu/100 μl (PUMA group, n=8), 10 mg/kg TMZ (TMZ group, n=8) and 2×108 pfu/100 μl Ad-PUMA + 10 mg/kg TMZ (combined group, n=8). Mice were killed after 20 days treatment.Tumor volume, inhibition rates and apoptotic index (AI) were measured, meanwhile, apoptotic tumor cells were detected by TUNEL technology respectively. The expression of MGMT mRNA and MGMT protein were revealed by the methods of RT-PCR and Western blot. Results According to the order: control group, AdPUMA group, TMZ group, combined group, tumor volumes were (3.68±0.09), (2.63±0.13), (2.13±0.07),(0.97±0.02) cm3 respectively (P＜0.05); the inhibitive rates were 0, 28.5 %, 42.1%, 73.6 % respectively and AI were (2.0±1.2) %, (11.4±2.6) %, (7.6±3.2) %, (20.6±8.6) % (P＜0.05). The results of Western blot and RT-PCR showed that MGMT mRNA and MGMT protein levels in TMZ group were higher than other groups (all P＜0.01). Conclusion Ad-PUMA combined with TMZ greatly enhances the sensitivity of human glioblastoma cells to TMZ and could effectively inhibit the proliferation and promote the apeptosis of glioblastoma cells, its mechanism was probably related Ad-PUMA promote apoptosis and inhibit MGMT expression.

Expression of PUMA, P53, Bax and Bcl-2 proteins in pancreatic ductal adenocarcinoma and the relevant clinical significance / 第二军医大学学报

Objective: To observe the expression of PUMA, P53, Bax and Bcl-2 protein in pancreatic ductal adenocarcinoma (PDA), and to investigate the role of PUMA in tumorigenesis of PDA and its relationship with P53, Bax and Bcl-2 protein expression. Methods: The expression of PUMA, P53, Bax and Bcl-2 protein was examined by immunohistochemical EnVision method in 65 PDA tissues and the corresponding adjacent tissues. Results: The positive rate of PUMA protein expression was 30.8% (20/65) in the PDA tissues, which was significantly lower than that in the corresponding adjacent tissues (49.2%[32/65], P<0.05). The expression of PUMA protein was significantly correlated with the tumor size and lymph node metastasis (P<0.05), but not with patient's age, sex, tumor location, differentiation degree, tumor stage or neural invasion. PUMA expression was negatively correlated with P53 and Bcl-2 expression (P = 0.019, P = 0.015), but was not correlated with Bax expression in PDA tissues. Conclusion: Decreased PUMA protein expression is correlated with tumor size and lymph node metastasis in PDA patients, suggesting that PUMA protein might be involved in the tumorigenesis and development of PDA. Detection of PUMA protein expression may be used for predicting the prognoses of PDA patients.