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OBJECTIVE@#The activation of Janus kinase (JAK) and signal transducers and activators of transcription (STAT) plays an important role in the prognosis and targeted therapy of ovarian high-grade serous carcinoma (HGSC). Utilizing simple and practicable technique, this study aimed to evaluate the activation of JAK/STAT signaling pathway in ovarian HGSC patients, and investigated the correlation between the activation of JAK/STAT signaling pathway and the prognosis of the HGSC patients.@*METHODS@#We performed immunohistochemistry of phosphorylated STAT3 (pSTAT3) and phosphorylated STAT5 (pSTAT5) on paraffin imbedded slides of 73 ovarian HGSC patients, and evaluated the expression level and range of both markers. According to the grading score of the immunostaining of pSTAT3 and pSTAT5, we divided the 73 ovarian HGSC cases into STAT3 low/high expression and STAT5 low/high expression groups, and analyzed the prognosis of the patients in different groups, in order to explore the relationship between the expression of pSTAT3 and pSTAT5 proteins and the prognosis of the HGSC patients.@*RESULTS@#Some of the ovarian HGSC cases showed high expression of pSTAT3 and pSTAT5 protein level, which was related to the poorer prognosis of the HGSC patients. There was a significant difference in the expression level of pSTAT3 and pSTAT5 between the patients with better prognosis (survival time ≥3 years) and poorer prognosis (survival time < 3 years). The patients with higher protein expression of pSTAT3, pSTAT5 or both markers might have poorer prognosis, with significant shorter progression-free survival time and overall survival time (P < 0.001).@*CONCLUSION@#Immunostaining of pSTAT3 and pSTAT5 proteins might be helpful to evaluate and predict the prognosis of the ovarian HGSC patients, and to identify the patients who might have higher chances to respond to the STAT inhibitors and anti-angiogenesis therapy.
Subject(s)
Humans , Prognosis , STAT5 Transcription Factor/metabolism , Neoplasms , Signal Transduction , ImmunohistochemistryABSTRACT
Objective: To investigate the clinicopathologic and molecular characteristics of fumarate hydratase (FH) deficient uterine leiomyoma. Methods: Eighty cases of FH deficient uterine leiomyoma were diagnosed from April 2018 to September 2022 in Department of Pathology, Peking University Third Hospital. Sanger sequencing of FH gene exons (exon 1-10) were performed on tumor tissues and matched non-tumor tissues/peripheral blood for all cases. FH immunohistochemistry were performed in 74 cases; S-(2-succino)-cysteine (2SC) were also detected by immunohistochemistry in five cases. Results: Patients' age ranged from 18 to 54 (36.0±7.5) years, with more than 60% exhibiting clinical symptoms of multiple and large leiomyomas (the median diameter was 70 mm). More than four histologic features, including staghorn vasculature, alveolar-pattern edema, bizarre nuclei, oval nuclei arranged in chains, prominent eosinophilic nucleoli with perinucleolar haloes and eosinophilic intracytoplasmic globules were observed in 98.5% (67/68) patients. The immunohistochemical sensitivity of FH and 2SC were 97.3% and 100%, respectively. Based on the Sanger sequencing results, the cases were divided into germline variant group (31 cases), somatic variant group (29 cases) and no variant group (20 cases). Sixty-nine percent (20/29) of the patients with FH germline variation had clear family history. Conclusions: Clinical features, histological morphology, FH and 2SC immunohistochemistry and Sanger sequencing have their own significance and limitations in differential diagnosis of FH deficient uterine leiomyoma. In clinical practice, the above information should be fully integrated and studied for accurate pathologic diagnosis and selection of patients with FH germline variation.
Subject(s)
Female , Humans , Adolescent , Young Adult , Adult , Middle Aged , Fumarate Hydratase/genetics , Uterine Neoplasms/pathology , Leiomyoma/pathology , Germ-Line Mutation , Diagnosis, Differential , Leiomyomatosis/pathology , Carcinoma, Renal Cell/diagnosisABSTRACT
Objective:To observe clinical effect of addition and subtraction therapy of Zidiantang to purpura nephritis in children （syndrome of blood fever） and regulatory action to immune inflammatory factors. Method:One hundred and twenty-five patients were randomly divided into control group （61 cases） and observation group （64 cases） by random number table. A total of 57 patients in control group completed the therapy （2 patients were falling off or missing visit and 2 was eliminated）， 59 patients in observation group completed the therapy （3 patients were falling off or missing visit and 2 was eliminated）. Both groups' patients got comprehensive measures of western medicine. Patients in control group got Xueniaoan capsule， 1 to 4 grains/time， 3 times/day. Patients in observation groups got addition and subtraction therapy of Zidiantang， 1 dose/day. The treatment was continued for 2 months and the follow up was recorded for a month. Purpura and urinalysis were recorded for every week. And disappearance of purpura， hematuria and proteinuria were compared for 3 months. Before treatment， and at the first， second and during the follow up， scores of 24 h urine protein quantification （24 hup） and syndrome of blood fever were graded. Levels of microalbuminuria （mAlb）， urinary <italic>β</italic><sub>2</sub>-microglobulin （<italic>β</italic><sub>2</sub>-MG）， cystatin C（CysC）， globulin A1（IgA1）， IgG， complement C3， Th17 cells， Treg cells， interleukin-17 （IL-17）， IL-21， IL-10 and transforming growth factor-<italic>β</italic><sub>1</sub> （TGF-<italic>β</italic><sub>1</sub>） were detected before and after treatment. Result:At the first month of treatment， disappearance rate of purpura in observation group was higher than that in control group （<italic>P</italic><0.05）. Before treatment， and at the first， second and during the follow up， disappearance rate of hematuria and albuminuria were higher than those in control group （<italic>P</italic><0.05）， and scores of 24 h urine protein quantification （24 hup） and syndrome of blood fever were lower than those in control group （<italic>P</italic><0.01）. Levels of mAlb， <italic>β</italic><sub>2</sub>-MG， CysC， IgA1， IgG， Th17， Th17/Treg， IL-17， IL-21 and TGF-<italic>β</italic><sub>1</sub> were lower than those in control group （<italic>P</italic><0.01）， and levels of C3， CD4<sup>+</sup>， Treg and IL-10 were higher than those in control group （<italic>P</italic><0.01）. Conclusion:On the basis of conventional western medicine treatment， addition and subtraction therapy of Zidiantang can promote the purpura to subside， improve the syndrome symptoms of blood fever， regulate the immune inflammatory reaction， reduce the inflammatory damage of kidney， thus reduce the hematuria and proteinuria， and play a role in protecting the renal function.
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Objective MicroRNAs (miRNA) play an important role in the development and progression of intervertebral disc degeneration (IDD), but the underlying mechanisms remain unclear. This study was to search for differentially expressed miRNAs and predict their target genes in the degenerative intervertebral disc tissue. Methods Data on the miRNA expression profile in the nucleus pulposus of the intervertebral disc were downloaded from the GEO database, involving nucleus pulposus samples from 3 cases of IDD and normal nucleus pulposus samples from another 3 patients with new traumatic lumbar fracture. Differentially expressed microRNAs were identified in the nucleus pulposus tissues of the IDD and normal control groups with the R Software, and the target genes significantly differentially expressed in the miRNAs were predicted using the miRWalk Software. The above target genes were enriched in the clusterProfiler package by GO biological function analysis and KEGG pathway analysis. Meanwhile, a protein-protein interaction network of the target genes was constructed with the STRING database and Cytoscape software, and the hub genes were identified. Based on the Pfirrmann grading of IDD, the subjects involved in the GSE23130 data were divided into a control group (≤grade 3, n = 15) and an IDD group (>grade 3, n = 8) followed by analysis of the expression levels of the hub genes. Results A total of 374 differentially expressed miRNAs were identified, 189 up-regulated and 185 down-regulated, with hsa-let-7b-5p most significantly down-regulated. Prediction of the 5 most significantly up- or down-regulated miRNAs showed the highest number of target genes in hsa-let-7b-5p, 85 in all, including GPAT4, E2F2, and PAK1. GO enrichment analysis manifested that these target genes were mainly involved in the biological processes of cell cycle G1/S phase transition and positive regulation of membrane-targeted proteins. The signaling pathways enriched in the target genes mainly included prolactin, insulin, p53 signaling pathways. Ten hub genes were identified by analysis of the PPI network, including CCND2, NRAS, E2F2, E2F6, STX3, CDCA8, RRM2, PPP2R2A, TXLNG and AKT2. The expression levels of CCND2, NRAS, E2F2, E2F6, STX3, CDCA8, RRM2, PPP2R2A and AKT2 in the degenerative intervertebral disc tissue were significantly higher than those in the control group (P < 0.05). Conclusion Significantly reduced expression of hsa-let-7b-5p in the nucleus pulposus tissue of IDD patients may play an important role in IDD by regulating its target genes CCND2, NRAS, etc.
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Objective Overexpressed inflammatory factors play an important role in the process of intervertebral disc degeneration. This study aimed to investigate the effect of iguratimod on the expression of inflammatory factors in degenerative intervertebral disc cells. Methods Sixty 8-12 weeks old SD rats were equally randomized into a compression (the tail compressed by external fixation) and a non-compression control group. The nucleus pulposus cells (NPC) of the degenerated intervertebral disc were isolated and treated with iguratimod at the concentrations of 0, 0.3, 3, 10, 20, and 30 μg/mL, followed by measurement of the contents of inflammatory factors and matrix metalloproteinases (MMP) secreted from the NPCs and determination of the effects of different concentrations of iguratimod on the expressions of inflammation-related genes in the NPCs by RT-PCR. Results After treatment with iguratimod at 3, 10, 20, and 30 μg/mL, the expression levels of IL-6 in the NPCs were (204.18 ± 6.96), (122.73 ± 9.38), (97.87 ± 7.81), and (86.31 ± 8.57) pg/mL, respectively, and those of TNF-α were (202.46 ± 7.84), (132.52 ± 11.4), (101.26 ± 10.38), and (96.89 ± 9.60) pg/mL, respectively, all decreased significantly in a concentration-dependent manner (P < 0.05). Meanwhile, the contents of MMP-2, MMP-3 and MMP-9 in the iguratimod-treated NPCs also showed remarkable concentration-dependent decreases (P < 0.05). Conclusion Iguratimod can effectively inhibit the expression of inflammatory factors in nucleus pulposus cells and block the progression of inflammatory response, which has provided a new idea for the treatment of degenerative intervertebral disc disease.
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Objective To investigate the clinical effect of porcelain fused for metal crown restoration after root canal therapy.Methods Totally 90 patients with pulpitis or periapical periodontitis from December 2013 to December 2015 had their clinical data analyzed,who underwent root canal therapy and were divided equally into an observation group and a control group.The observation group took porcelain endocrowns repair and the control group applied ceramic crown.The two groups were compared on the restoration integrity,edge sealing,residual rate,gum,food impaction and color 3 months,6 months and 1 a after treatment.Results The observation group behaved significantly better than the control group in the edge sealing and restoration integrity (P＜0.05).There were significant differences between the residual rates,gum and food impaction in the two groups (P＜0.05).Conclusion Porcelain endocrowns repair gains advantages over ceramic crown repair in edge sealing,integrity,gum,food impaction and color after root canal therapy,though its long-term effect needs further investigation.
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Objective To investigate the clinical effect of porcelain fused for metal crown restoration after root canal therapy.Methods Totally 90 patients with pulpitis or periapical periodontitis from December 2013 to December 2015 had their clinical data analyzed,who underwent root canal therapy and were divided equally into an observation group and a control group.The observation group took porcelain endocrowns repair and the control group applied ceramic crown.The two groups were compared on the restoration integrity,edge sealing,residual rate,gum,food impaction and color 3 months,6 months and 1 a after treatment.Results The observation group behaved significantly better than the control group in the edge sealing and restoration integrity (P＜0.05).There were significant differences between the residual rates,gum and food impaction in the two groups (P＜0.05).Conclusion Porcelain endocrowns repair gains advantages over ceramic crown repair in edge sealing,integrity,gum,food impaction and color after root canal therapy,though its long-term effect needs further investigation.
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Objective:To observe whether pretreatment with Pam3CSK4,a TLR2 agonist,could decrease the inflammation response in kidney from mice with systemic MRSA infection,and to investigate the mechanism of the attenuation of inflammation with Pam3CSK4 pretreatment. Methods:BALB/c mice were pretreated with Pam3CSK4 (10 μg/100 μl/each mouse) or PBS via tail vein once daily for two consecutive days. All mice were infected with live MRSA (ATCC43300) at 2×107 CFU/each mouse (via tail vein) 24 h after the second treatment. The levels of cytokines in kidney were measured by ELISA and real-time PCR,respectively. The relative expression of TLR2,IRAKs etc. were detected by real-time PCR. Western blot was performed to detect the phosphorylation of NF-κB, the expression of IRAK-M and A20,respectively. Results:The level of TNF-α,IL-6,IL-1β,CCL3 and IFN-γ in renal tissue from mice pretreated with Pam3CSK4 was decreased significantly compared with that from PBS-treated mice,respectively. Pam3CSK4 pretreatment down-regulated the relative expression of TLR2, inhibited the expression of IRAK-1 and the phosphorylation of NF-κB post infection. The expression of IRAK-M,one of the negative regulators in TLRs signaling pathway was increased significantly in renal tissue from Pam3CSK4-treated mice post infection. Conclusion:Pam3CSK4 pretreatment attenuated the inflammation response in kidney from mice with systemic MRSA infection,and these attenuation is related with up-regulation of IRAK-M.
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Objective:To observe whether pretreatment with Pam3CSK4,a TLR2 agonist,could decrease the inflammation response in kidney from mice with systemic MRSA infection,and to investigate the mechanism of the attenuation of inflammation with Pam3CSK4 pretreatment. Methods:BALB/c mice were pretreated with Pam3CSK4 (10 μg/100 μl/each mouse) or PBS via tail vein once daily for two consecutive days. All mice were infected with live MRSA (ATCC43300) at 2×107 CFU/each mouse (via tail vein) 24 h after the second treatment. The levels of cytokines in kidney were measured by ELISA and real-time PCR,respectively. The relative expression of TLR2,IRAKs etc. were detected by real-time PCR. Western blot was performed to detect the phosphorylation of NF-κB, the expression of IRAK-M and A20,respectively. Results:The level of TNF-α,IL-6,IL-1β,CCL3 and IFN-γ in renal tissue from mice pretreated with Pam3CSK4 was decreased significantly compared with that from PBS-treated mice,respectively. Pam3CSK4 pretreatment down-regulated the relative expression of TLR2, inhibited the expression of IRAK-1 and the phosphorylation of NF-κB post infection. The expression of IRAK-M,one of the negative regulators in TLRs signaling pathway was increased significantly in renal tissue from Pam3CSK4-treated mice post infection. Conclusion:Pam3CSK4 pretreatment attenuated the inflammation response in kidney from mice with systemic MRSA infection,and these attenuation is related with up-regulation of IRAK-M.
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A B/S framework -based library entrance examination system was developed for new students using the PHP language and SQL server database according to their unsatisfactory training results, which can maintain the in-formation of entrance examination-involved students and examination papers, automatic generating and marking of examination papers. The new students were registered to a regular reader after they passed the examination, and could thus have the right to entry the library, select a seat, loan books or journals, use a computer, and participate in library-held culture activities. The new students could understand the library rules and regulations, control their behaviors against library rules and regulations, and rapidly place themselves in the study environment of library.
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<p><b>OBJECTIVE</b>To investigate the expression profiles of bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) during the development of mouse adipose tissue.</p><p><b>METHODS</b>The total RNA was extracted for real-time PCR for amplification of BAMBI mRNA from the suprascapular brown adipose tissue (BAT) and subcutaneous (inguinal) and visceral (gonadal) white adipose tissue (sWAT and vWAT, respectively) of mice at various embryonic and postnatal stages, as well as from isolated primary preadipocytes during differentiation.</p><p><b>RESULTS</b>In BAT, BAMBI mRNA levels exhibited a transient increase, peaking at day 0 (D0) and declined thereafter. sWAT and vWAT could be isolated from mice from postnatal D21 onwards, in which BAMBI mRNA levels were the highest and decreased at 8 weeks and 6 months. BAMBI mRNA levels were also significantly reduced in primary preadipocytes isolated from vWAT after induced differentiation. BAMBI mRNA expression level was higher in vWAT than in sWAT and BAT at the same developmental stages.</p><p><b>CONCLUSION</b>BAMBI is differentially expressed in different adipose tissues and developmental stages, which supports the hypothesis that BAMBI plays a pivotal role in the development of adipose tissues.</p>
Subject(s)
Animals , Mice , Adipocytes , Metabolism , Adipose Tissue , Metabolism , Bone Morphogenetic Proteins , Metabolism , Cell Differentiation , Membrane Proteins , Metabolism , Phosphorylation , RNA, Messenger , Real-Time Polymerase Chain Reaction , Signal TransductionABSTRACT
This study is to investigate the effects of indirubin on ATP-induced immune responses of macrophages. For this, neutral red dye uptake method was used to test phagocytosis, MTT assay was used for measuring cell death, and reactive oxygen species (ROS) was tested with fluorescent probe DHE. The data showed that extracellular ATP attenuated phagocytosis, induced cell death and increased ROS production, and these effects were restored by pre-treating with indirubin. This result suggested that indirubin blockade the effects of ATP on macrophages, because extracellular ATP-induced effects are dependent on P2 receptors, in particular P2X7 receptors. Furthermore, the effects of indirubin on the activation of P2 receptors were tested, in particular P2X7 receptors. The data showed that indirubin significantly decreased ATP-induced, P2 receptors mediated intracellular Ca2+ concentration ([Ca2+]i) rise and inhibited P2X7 receptor-based ethidium bromide (EB) dye uptake. These results suggested the inhibitory effects of indirubin on the activation of P2X7 receptors, which may underlying the effects on ATP induced ROS production, phagocytosis attenuation and cell death of macrophages.
Subject(s)
Animals , Male , Rats , Adenosine Triphosphate , Pharmacology , Calcium , Metabolism , Cell Death , Indoles , Pharmacology , Macrophages , Cell Biology , Metabolism , Physiology , Phagocytosis , Rats, Wistar , Reactive Oxygen Species , Metabolism , Receptors, Purinergic P2X7 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of MDC1 gene silencing by RNA interference on the radiosensitivity of human esophageal carcinoma cell line ECA109.</p><p><b>METHODS</b>The vectors containing short hairpin RNA (shRNA) targeting MDC1 gene (pMDC1-shRNA) were cotransfected with pPACKH1-lentivector packaging system into 293T cells to package the lentivirus particles. Forty-eight hours after the transfection with specific or control lentiviral vectors, the stable integrants were selected using copGFP reporter gene; real-time RT-PCR and Western blotting were used to detect the expression levels of MDC1 mRNA and protein in the transfected ECA109 cells, respectively. The cell cycle distribution was measured with flow cytometry at 12, 24 and 48 h after a 5 Gy irradiation, and the radiosensitivity of esophageal carcinoma cell was evaluated by clone formation array.</p><p><b>RESULTS</b>Sequence analysis confirmed correct insertion of MDC1-shRNA construct into pSIH1-H1-copGFP. The percentage of G2/M phase ECA109/ MDC1 cells was lower than that of ECA109 and ECA109/negative cells. The value of D0, SF2 and Dq of ECA109/ MDC1 cells were 1.88 Gy, 0.84 and 1.20, respectively, lower than those of ECA109 cells (3.06 Gy, 0.91 and 1.59) and those of ECA109/negative cells (2.90 Gy, 0.89 and 1.47).</p><p><b>CONCLUSION</b>RNA interference can inhibit MDC1 gene expression and enhance the radiosensitivity of ECA109 cells in vitro.</p>
Subject(s)
Humans , Carcinoma, Squamous Cell , Genetics , Pathology , Cell Line, Tumor , Esophageal Neoplasms , Genetics , Pathology , Gene Expression Regulation, Neoplastic , Genetic Vectors , Nuclear Proteins , Genetics , RNA Interference , RNA, Small Interfering , Genetics , Radiation Tolerance , Genetics , Trans-Activators , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To explore the distribution of lymph node metastases, to analyze the cliniopathologic factors of thoracic esophageal carcinoma after curative resection, and to provide the criteria of irradiated region delineation in radiotherapy for esophageal carcinoma.</p><p><b>METHODS</b>The clinicopathological data of 763 patients who underwent esophagecotomy from Jun 2002 to Jun 2006 were retrospectively analyzed. The regularity of lymph node metastases of thoracic esophageal cancer and clinicopathological factors were stratified and analyzed with SPSS13.0 software.</p><p><b>RESULTS</b>Of the 763 patients, a total of 5846 lymph nodes were dissected with an average of 7.7 lymph nodes in each case. Metastatic lymph nodes were 711, the ratio of metastatic lymph node was 12.2%, and 297 patients had lymph node involved, the lymph node metastasis rate was 38.9%. The metastatic lymph nodes of upper-thoracic esophagus were mainly observed in the supraclavicular and paratracheal regions (P < 0.05), the metastatic lymph nodes of middle-third thoracic esophagus were bidirectional, and those of the lower-third thoracic esophagus mainly metastasized to the regions adjacent to the esophagus, gastric cardia and gastric artery (P < 0.05). Both the metastasis ratio and rate of lymph nodes adjacent to the gastric artery in the lower-thoracic esophageal cancer were significantly higher than those in the middle-third and upper-third thoracic esophageal cancers (P = 0.007, P = 0.001). The multiple factors logistic regression analysis showed that tumor length, depth of tumor invasion, vascular tumor emboli and distant metastasis were major factors for lymphatic metastasis (P < 0.01). For the whole group of patients the lymph node metastatic rate was 28.5% in upper-thoracic esophageal cancer, significantly lower than 38.8% of the lower-thoracic esophageal cancer (P = 0.039) and 43.4% in the middle-thoracic esophageal cancer (P = 0.010). However, the lymph node metastatic rates were 37.0%, 37.9% and 41.4% in the upper-, middle- and lower-thoracic esophageal cancers of the 592 cases receiving left chest notches, with a non-significant difference among them (P = 0.715).</p><p><b>CONCLUSION</b>The lesion length, depth of tumor invasion, vascular tumor embolus and distant metastasis are the most important parameters for lymph node metastases. Operative modes have obvious influence on the distribution of regional lymph node metastases. Therefore, in the clinical management, a postoperative prophylactic radiotherapy may be selected according to the tumor length, depth of tumor invasion, vascular tumor embolus and distant lymph node metastasis.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adenocarcinoma , Pathology , Radiotherapy , General Surgery , Carcinoma, Squamous Cell , Pathology , Radiotherapy , General Surgery , Esophageal Neoplasms , Pathology , Radiotherapy , General Surgery , Follow-Up Studies , Lymph Node Excision , Methods , Lymph Nodes , Pathology , General Surgery , Lymphatic Irradiation , Lymphatic Metastasis , Neoplasm Invasiveness , Neoplasm Staging , Neovascularization, Pathologic , Pathology , Retrospective Studies , Small Cell Lung Carcinoma , Pathology , Radiotherapy , General Surgery , Survival RateABSTRACT
<p><b>OBJECTIVE</b>To investigate P2Y purinergic receptor activated PI-3K/Akt signaling pathway in the regulation of growth and invasion of prostate cancer in vitro.</p><p><b>METHODS</b>Western blot was used to detect phosphorylation of Akt (a downstream target molecule of PI-3K) by P2Y receptor agonist in 1E8 cells (a highly metastatic subclone derived from PC-3 prostatic cancer cell line). Cell counts, flow cytometry, Matrigel invasion assay, wound healing assay and gelatin zymography were used to detect changes of biological behaviors of 1E8 cells after P2Y receptor activation.</p><p><b>RESULTS</b>AMP-PNP, one non-hydrolysis ATP analogue and P2Y receptor agonist, induced significant phosphorylation of Akt in a time- and dose-dependent manner in IE8 cells. LY294002, a specific inhibitor of PI-3K, effectively blocked Akt phosphorylation induced by AMP-PNP. Continuous exposure to AMP-PNP induced significant growth inhibition of 1E8 cells (inhibition rate at 50.2% at the 8th day), and this inhibition was mainly due to an arrest at S phase of the cell cycle (the S phase fraction of AMP-PNP treated cells was 22.3% higher than that of the control). Application of LY294002 did not reverse the growth inhibition effect of AMP-PNP. Matrigel invasion assay showed that AMP-PNP stimulation increased invasive ability of 1E8 cells, and this effect was effectively blocked by LY294002. No significant changes in the activation of MMP-2 and MMP-9 were detected by gelatin zymography, although wound healing assay showed 21.2% increase in cell migration after AMP-PNP treatment.</p><p><b>CONCLUSIONS</b>PI-3K/Akt signaling pathway participates in P2Y receptor-stimulated prostate cancer invasion by enhancing cell motility, rather than up-regulating MMP-2 and MMP-9 activities. PI-3K signaling pathway plays an important role in prostate cancer proliferation, but is not involved in P2Y receptor mediated growth inhibition.</p>
Subject(s)
Animals , Humans , Male , Mice , Adenylyl Imidodiphosphate , Pharmacology , Apoptosis , Cell Line, Tumor , Cell Movement , Cell Proliferation , Chromones , Pharmacology , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Mice, Nude , Morpholines , Pharmacology , Neoplasm Invasiveness , Phosphatidylinositol 3-Kinases , Metabolism , Phosphorylation , Prostatic Neoplasms , Metabolism , Pathology , Proto-Oncogene Proteins c-akt , Metabolism , Purinergic P2 Receptor Agonists , S Phase , Signal TransductionABSTRACT
<p><b>OBJECTIVE</b>To observe the expression of DNA damage checkpoint mediator 1 (MDC1) and p53-binding protein 1 (53BP1) at both mRNA and protein levels and their significance in different human esophageal cancer cell lines.</p><p><b>METHODS</b>In 3 human esophageal carcinoma cell lines, TE-1, TE-13 and Eca109 cells, the expressions of MDC1 and 53BP1 mRNA were detected with RT-PCR, and MDC1 and 53BP1 protein expressions were measured with immunohistochemistry, indirect immunofluorescence and Western blotting, respectively.</p><p><b>RESULTS AND CONCLUSIONS</b>MDC1 and 53BP1 expressions were observed for the first time in human esophageal carcinoma cell lines TE-1,TE-13 and Eca109 cells, at both the mRNA and protein levels. The expressions of MDC1 and 53BP1 proteins may be implicated in the radiosensitivity of human esophageal carcinoma.</p>
Subject(s)
Animals , Humans , Blotting, Western , Cell Line, Tumor , DNA Damage , Genetics , Esophageal Neoplasms , Genetics , Metabolism , Pathology , Flow Cytometry , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Genetics , Metabolism , Nuclear Proteins , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Trans-Activators , Genetics , Metabolism , Tumor Suppressor p53-Binding Protein 1ABSTRACT
The recent studies have suggested that matrix metalloproteinases(MMPs)are close associated with the instability of atherosclerotic plaques,the formation and development of intracranial aneurysm,ischemic stroke and hemorrhagic transformation.The study and application of MMP inhibitor may become a new approach in the treatment of cerebrovascular diseases.
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<p><b>OBJECTIVE</b>To observe the effect of RNA interference on CHK1 and CHK2 expression and change of G2/M phase arrest in esophageal carcinoma cells after irradiation.</p><p><b>METHODS</b>Four sequences short hairhip RNA (shRNA) of each CHK1 and CHK2 genes were constructed and connected with vector of pENTR/U6 plasmid, respectively, and then transfected into Eca109 cells with lipofectamine 2000 reagent. Protein and mRNA expression of CHK1 and CHK2 genes were detected with Western blotting and RT-PCR, respectively. Cell cycling was measured by flow cytometry after 5 Gy irradiation. Cell survival rate after 5 Gy irradiation was evaluated by clonegenetic assay.</p><p><b>RESULTS</b>Four shRNA vector each of CHK1 and CHK2 genes were successfully constructed and transfected into Ecal09 cells, respectively. Protein expression of CHK1 and CHK2 were obviously decreased. Their mRNA expressions were also decreased after transfected with shRNA of CHK1 and CHK2. Arrest of G2/M stage in Eca109 cells were obviously decreased only in cells transfected with CHK1 shRNA but not with CHK2 shRNA at 12 h after 5 Gy irradiation. In first progeny Eca109 cells transfected with CHK1 and CHK2 shRNA, expression of CHK1 and CHK2 protein was also decreased. The level of phosphorylated CHK2-T68 expression was decreased at 1 h after 5 Gy irradiation, and at 72 h only transfected with CHK2 shRNA but not with CHK1 shRNA. Phosphorylation level of CHK1-S345 was not increased after transfected with CHK1 or CHK2 shRNA, but arrest of G2/M stage still remained at 12 h after 5 Gy irradiation and at 72 h accordingly. The cell survival rate was decreased in Eca109 cells transfected with CHK1 or CHK2 shRNA after 5 Gy irradiation.</p><p><b>CONCLUSION</b>After transfected with shRNA of CHK1 or CHK2, their expressions of mRNA and protein in Ecal09 cells are markedly inhibited and this inhibition effect can be observed in their first progeny cells and at least hold for 3 days. Arrest of G2/M phase can be reduced after irradiation when teansfected with shRNA of CHK1 and the radiosensitivity of Ec109 cells can be increased.</p>