RESUMEN
Objective:To investigate the differences of integral alpha 3 (ITGA3) mRNA and protein expressions in gliomas of different grades and different cell lines, and gliomas tissues of different clinical and molecular characteristics, and evaluate their prognostic values in brain glioma patients.Methods:(1) ITGA3 mRNA expression data in the brain gliomas and clinical data of these glioma patients were obtained from The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA). The differences of ITGA3 mRNA expressions in glioma patients with different gender, ages, WHO grading, isocitrate dehydrogenase 1 ( IDH) mutation statuses, and 1p/19q co-deletion statuses were compared. Kaplan-Meier method was used to plot and compare the survival curves of patients with ITGA3 mRNA high expression and ITGA3 mRNA low expression. Receiver operating characteristic (ROC) curve was used to analyze the predictive efficiency of ITGA3 mRNA expression in survival rate of gliomas. Univariate and multivariate Cox regression analyses were used to explore the independent influencing factors for prognoses in glioma patients. The independent influencing factors for prognosis were used to construct nomograms and the calibration diagram was used to verify the reliability of nomograms in predicting the prognoses of these patients. (2) Intracellular localization of ITGA3 and ITGA3 protein expression in low- and high-grade gliomas were determined by on-line database of Human Protein Atlas (HPA). (3) Brain glioma cells U87, U118, U251 and human astrocytes SVG were cultured in vitro, and the ITGA3 mRNA and protein expressions in cells were detected by Western blotting and reverse transcription (RT)-PCR, respectively. Results:(1) In TCGA database, the ITGA3 mRNA expressions in gliomas of WHO grading II, III and IV increased successively, with significant differences (P<0.05). In CGGA database, the ITGA3 mRNA expression in glioma of WHO grading IV was statistically higher than that in glioma of WHO grading II and III ( P<0.05). In TCGA and CGGA databases, the ITGA3 mRNA expressions in glioma patients aged ≤40 years and >40 years, patients with IDH wild-type and IDH mutation, and patients with chromosome 1p/19q deletion and chromosome 1p/19q non-deletion were statistically different ( P<0.05). The survival rate of patients with low ITGA3 mRNA expression was significantly higher than that of patients with high ITGA3 mRNA expression, no matter in low-grade glioma, glioblastoma, or entire glioma samples ( P<0.05). ROC curve showed that, in TCGA database, the area under the curve (AUC) of ITGA3 mRNA in predicting 1, 3, and 5 years survival was 0.791, 0.786, and 0.708 in glioma patients; in CGGA database, the AUC of ITGA3 mRNA in predicting 1, 3, and 5 years survival was 0.661, 0.667, and 0.659. Multivariate Cox regression analysis showed that, in TCGA database, age, WHO grading, IDH mutation, chromosome 1p/19q deletion and ITGA3 mRNA expression ( HR=1.018, 95%CI: 1.006-1.030, P=0.0.003) were independent influencing factors for prognoses of glioma patients ( P<0.05); and in CGGA database, WHO grading, IDH mutation, chromosome 1p/19q deletion, and ITGA3 mRNA expression ( HR=1.445, 95%CI: 1.132-1.844, P=0.003) were independent influencing factors for prognoses of glioma patients ( P<0.05). Nomograms showed that age had the greatest influence in survival, followed by ITGA3 mRNA expression. Calibration plots showed that nomogram was reliable in predicting 1-, 3-, and 5-year survival in glioma patients. (2) Immunofluorescence localization showed that ITGA3 protein mainly aggregated in cell membrane and vesicles. Immunohistochemical staining showed that the ITGA3 protein expression in high-grade glioma tissues was obviously higher than that in low-grade glioma tissues. (3) The results of RT-PCR and Western blotting revealed that the ITGA3 mRNA and protein expressions in glioma cell lines U87, U118 and U251 were significantly higher than those in SVG cells ( P<0.05). Conclusion:The ITGA3 mRNA and protein expression levels are correlated with the malignant degrees of glioma; patients with ITGA3 mRNA low expression tend to have a high overall survival; ITGA3 mRNA expression can be used as an index to evaluate the prognoses of glioma patients.
RESUMEN
Objective To investigate the effects of inhibitor of growth 5 (ING5) gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells.Methods The eukaryotic ING5-expressing plasmid and GFP-empty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment.Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability (P<0.05).Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.
RESUMEN
Objective To detect the expression of micro RNA (miR)-433 and histone deacetylase 6 (HDAC6) in glioma tissues and investigate the effect of miR-433 on cell proliferation and invasion of human glioma cell line U251. Methods (1) Forty-two glioma samples, collected from patients accepted surgical resection and conformed by pathology in our hospital from January 2010 to December 2014, and 13 healthy brain tissues, collected from patients accepted surgery for craniocerebral trauma at the same time period, were used in our study; reverse transcription (RT)-PCR was used to detect the mRNA expression levels of miRNA-433 and HDAC6 in the glioma samples and brain tissues. (2) Human glioma cell line was routinely cultured and divided into blank control group, nonsense sequence control group and miRNA-433 mimics group;cells in the later two groups were transfected with nonsense sequences or miRNA-433 mimics, and cells in the blank control group did not give any treatment;the mRNA expression levels of miRNA-433, P21 and HDA C6 in these 3 groups were detected by RT-PCR;the cellular viability was measured by CCK-8 assay;flow cytometry was used to monitor the changes of cell cycle and apoptosis; cell invasion was evaluated by Transwell assay; HDAC6 protein expression was detected by Western blotting. (3) Wide-type (WT)HDAC63'-UTR and mutant type (MUT)HDAC63'-UTR luciferase report vectors were established; miR-433 mimics+WT HDAC63′-UTR and nonsense sequences+WT HDAC63'-UTR were transfected into the U251 cells, and dual-luciferase experiment was used to detect the fluorescence intensity of the cells; miR-433 mimics+MUT HDAC63'-UTR and nonsense sequences+MUT HDAC63'-UTR were transfected into the U251 cells, and dual-luciferase experiment was used to detect the fluorescence intensity of the cells. (4) U251 cells were divided into nonsense sequence control group, HDAC6 expression plasmids group and HDAC6 siRNA group, and nonsense sequences, HDAC6 expression plasmids or HDAC6 siRNA were transfected respectively; RT-PCR was used to detect the P21 and HDAC6 mRNA expressions and miRNA-433 expression; U251 cells were divided into miR-433 mimics group and miR-433 mimics+HDAC6 expression plasmids group, and miR-433 mimics or miR-433 mimics+HDAC6 expression plasmids were transfected, respectively, and one-5 d after that, CCK-8 was used to detect the cellular viability. Results (1) The miRNA-433 expressions gradually increased and HDA C6 mRNA expressions gradually decreased in the high-grade gliomas, low-grade gliomas and normal brain tissues, and significant differences were noted among each two groups (P<0.05);the miRNA-433 expression was negatively correlated with HDA C6 mRNA expression in the glioma tissues (r=0.829, P=0.000). (2) As compared with blank control group and nonsense sequence control group, miRNA-433 mimics group had significantly higher miRNA-433 and P21 mRNA expressions, cell percentage at G0/G1 stage, and apoptotic rate (P<0.05), and had statistically lower HDAC6 mRNA expression, cellular viability on 2-5 d of culture, number of transmembrane cells and HDAC6 protein expression (P<0.05). (3) The luciferase activity in cells from miR-433 mimics+WT HDAC63'-UTR group was significantly lower as compared with that in the nonsense sequences+WT HDAC63'-UTR group (P<0.05);the luciferase activity in cells from miR-433 mimics+MUT HDAC63'-UTR group and nonsense sequences+MUT HDAC63'-UTR group showed no significant differences (P>0.05). (4) The HDA C6 mRNA expressions were gradually increased, and P21 mRNA expressions were gradually decreased in the HDAC6 siRNA group, nonsense sequence control group, and HDAC6 expression plasmids group, with significant differences (P<0.05);on 2-5 d of culture, the cellular viability in the miR-433 mimics+HDAC6 expression plasmids group was significantly higher than that in the miR-433 mimics group (P<0.05). Conclusions The miRNA-433 expression level is low in human glioma tissues;miRNA-433 over-expression may inhibit the cell activity and promote cell apoptosis of glioma cell line U251 in vitro via inhibiting the HDAC6 expression.
RESUMEN
OBJECTIVE:To prepare transferrin modified paclitaxel-loaded liposome(TF-PTX-LP),and to study the tumor in-hibition effect. METHODS:TF-PTX-LP was prepared by thin-film method,and morphology of TF-PTX-LP was observed. Qualita-tive and quantitative investigation were used to value the uptake efficiency of TF-LP and LP by HepG2 cells. The proliferation inhi-bition rate of HepG2 cells was investigated after treated with PTX,PTX-LP and TF-PTX-LP for 24,48 and 72 h. Tumor spheres were prepared by using HepG2 cells. Effects of normal saline,PTX,PTX-LP and TF-PTX-LP on the volume of tumor spheres were investigated after 0,1,2,4,5,6 and 7 d treatment. HepG2 tumor-bearing nude mice model was induced. Inhibitory effects of normal saline,PTX,PTX-LP and TF-PTX-LP(8.5 mg/kg by PTX)on transplantable tumor of tumor-bearing nude mice were in-vestigated. RESULTS:TF-PTX-LP showed uniform spherical shape,with particle size of 100-120 nm. The fluorescence intensity of HepG2 cells treated with TF-LP was stronger than that treated with LP(P<0.01). Compared with PTX and PTX-LP,TF-PTX-LP showed higher proliferation inhibition rate(P<0.01). Compared with normal saline,PTX and PTX-LP,tumor spheres were small-er in volume after treated with TF-PTX-LP,and inhibition rate of tumor was higher in tumor-bearing nude mice;there were statisti-cal significance after treated for 6,7 d(P<0.01). The proliferation inhibition rate and tumor spheres volume changed in time-de-pendent manner. CONCLUSIONS:TF-PTX-LP which owns good tumor inhibition effect is prepared successfully.
RESUMEN
Objective To detect the expression of mitochondrial dynamics proteins (Mfn2 and Drp1) in thyroid squa?mous carcinoma cell line SW579 and the effects of Mitochondrial division inhibitor, Mdivi-1, on proliferation, apoptosis and invasion of SW579. Methods In SW579 and Nthy-ori 3-1 cell lines, the expression levels of Mfn2 and Drp1 were deter?mined by western blot while the transcription level of Mfn2 and Drp1 mRNA were measured by RT-PCR. Then, SW579 cells were divided into control group (DMSO, 0.1%) and Mdivi-1 low, medium and high dose groups (Mdivi-1 of 15,30 and 45μmol/L were incubated with cells for 16 hours respectively). Then the ability of cell proliferation was detected using MTT assay, the mitochondrial membrane potential was determined by fluorescence spectrophotometer, the expression levels of cy?tochrome C and Caspase-3 were quantified by Western blot and the transcription level of the Cyt C and Caspase-3 mRNA were determined by RT-PCR. The ability of invasion in each group was measured with Transwell assays. Results Com?pared with Nthy-ori 3-1, the mRNA transcription and protein expression levels of the Mfn2 was remarkably decreased, while the mRNA transcription and protein expression of the Drp1 was significantly increased in SW579 cells (P<0.01). Compared with control group, the cell survival rates and mitochondrial membrane potential of SW579 were decreased dramat?ically (P<0.01). The mRNA transcription and protein expression of the cytochrome C and Caspase-3 were increased dra?matically (P<0.01) and the capability of invasion was markedly decreased in all the Mdivi-1 groups in a dosage dependent manner compared with those in control groups (P<0.01). Conclusion Abnormal mitochondrial dynamics may be involved in thyroid squamous cell carcinoma SW579 cells;Mdivi-1 can inhibit the cell proliferation and invasion as well as induce apoptosis.
RESUMEN
Objective To investigate the correlation between the single nucleotide polymorphism (SNP) of tumor necrosis factor (TNF-α) gene promotor-238 and hepatitis B virus (HBV) reactivation in patients with malignant tumors after chemotherapy.Methods Polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP) was used to detect the SNPat TNF-α-238 site among 100 malignant tumor patients with HBV infection.HBV-DNA levels in patients were detected before and after chemotherapy.HBV reactivation was defined as that HBV-DNA level greater than 10-fold increase compared with before chemotherapy or higher than 1 × 109 logcopies/ml.Results The quantification of HBV-DNA was higher after chemotherapy than before chemotherapy [(3.02±0.68) logcopies/ml vs.(2.49±0.23) logcopies/ml,t=-7.383,P=0.000].Among the patients with malignant tumor and HBV infection,the genotype frequency of G/A was higher in HBV reactivation group than in non-reactivation group after chemotherapy [27.3％ (6/22) vs.3.8％ (3/ 78),x2 =11.499,P=0.001].Conclusions HBV reactivation is associated with TNF-α-238 gene polymorphism in malignant tumor patients with HBV infection after chemotherapy.
RESUMEN
Objective To explore the anti-apoptotic mechanism of Roux-en-Y gastric bypass (RYGB) through exam?ine the postoperative change of adiponectin levels and expressions of pancreatic islets relative apoptotic protein. Methods Sixty SD rats were randomly allocated to RYGB group (n=20), type 2 diabetes mellitus group (T2DM, n=20) and normal con?trol group (NC, n=20). Rats in the NC group were fed with normal diet. In order to make type 2 diabetic rat models, the rats in the T2DM and RYGB groups were fed with high fat diet (22.19 kJ/g) combined with administration of intraperitoneal strep?tozotocin injection (STZ, 30 mg/kg) on the 13th day of high fat diet. RYGB operation were performed in RYGB group and sham-operation were performed in the T2DM and NC groups when diabetic model was contructed. Rats were weight preoper?atively and at the 7th, 14th, 21st days after operations. Fasting plasma glucose and adiponectin (ELISA) were measured preoper?atively and at 21st day postoperatively. Protein expressions of Bcl-2,Caspase 8 and Caspase 9 in pancreatic islets were ex?amined by immunohistochemistry at the 21st day postoperatively. Results Body weights do not vary significantly among three groups preoperatively. Compared to rats in the NC group, fast plasma glucose level was higher but adiponectin was low?er in rats in RYGB and T2DM groups. Body weights of rats in RYGB group decreased significantly compared to those of rats in NC and T2DM groups postoperatively. Compared to rats in T2DM group, fasting glucose level was lower while adiponectin concentrations was higher in rats in RYGB group but no differences of these parameters were seen in rats in NC group at the 21st day postoperatively. Expression of Bcl-2 in RYGB group was significantly elevated while expressions of Caspase 8 and Caspase 9 were significantly decreased compared to those in T2DM group postoperatively. Conclusion Adiponectin levelswas elevated;expressions of Bcl-2 was increased;expressions of Caspase 8, Caspase 9 were decreased upon RYGB opera?tion in T2DM model. RYGB might reduce pancreatic islets apoptosis through mitochondrial pathway.
RESUMEN
Purpose To detect the expression of osteopontin(OPN) and minkine(MK) protein in paraffin-embedded samples obtained from patients with infiltrating breast carcinoma and to analyze the relationships between their expression and clinical feature and prognosis in order to provide valuable biomarkers for the treatment and prognosis of infiltrating breast carcinoma.Methods The expression of OPN and MK was measured in 90 infiltrating breast cancer tissues and 30 hyperplastic disease of the breast by immunohistochemical PV-9000 two-step method.Results Immunohistochemical results showed that the positive rates of OPN and MK in infiltrating breast carcinoma were 71.11% (64/90) and 78.89 (71/90), respectivelyy, which were significantly higher than that in hyperplastic diseases of breast [30.00% (9/30) and 46.67% (14/30)](P0.05).The expression of MK was significantly correlated with tumor size,axillary lymph node metastasis,recurrence of tumor , the expression of p185 and TNM stage(P0.05).There was a significantly positive relationship between OPN and MK expression (r=0.274,P=0.009). TTP and OS in patients with OPN and MK positive expression were shorter than those with OPN and MK negative expression (P<0.05).Conclusions OPN and MK′s expression may correlate with the progression of breast cancer.OPN and MK′s detection may provide a theoretical basis for the diagnosis and treatment of infiltrating breast carcinoma.
RESUMEN
Tension-free hernioptasty is a major method for treating inguinal hernia, characterizing by less wound, tension-free incision, sample processing, less dissection and separation, mild pain, rapid recovery, and short length of stay. In particular, transabdominal preperitoneal inguinal hernioplasty is considered as the most ideal method at physiologicoanatomical level. However, the effect of patch materials on postoperative recovery is generally ignored for a long time. Recent studies demonstrate that patch weight and mesh diameter are important factors for abdominal wall repairing and reconstruction. Polypropylene patch, characterizing by strong stability, high intensity, great unreactiveness, and sample processing, has become a major material for hernioptasty. However, there are still some problems of clinical application, including that polypropylene patch may induce intensive foreign body reaction and chronic inflammation as well as compliance decreasing of abdominal wall. Therefore, polypropylene patch should be chosen according to patient requirement, i.e., light, soft, and wide-aperture patch, so as to reduce onset of complication. Nerve and ligament tissues could be clearly observed from light and soft patch, and this might and prevent hurt by mistake, improve repairing operability, enhance correct putting of patch, and decrease incidence of pain. Furthermore, the structure was beneficial for in-growth of fiber tissue, longitudinal and transversal movement induced by body position alternation and muscle contraction, and reduction of discomfortableness.
RESUMEN
Objective To investigate the relationship between the expression of p53, ki-67 proteins and clinicopathology of human thyroid carcinoma. Methods Using SP immunohistochemical methods, the expression of p53 and ki-67 proteins was detected in 55 cases of thyroid carcinoma. Result The positive rate of p53 protein expression was 52 7%(29/55) and ki-67 proliferative index was(11 07?3 84)% in the thyroid carcinoma. The positive rate of p53 protein expression and ki-67 proliferative index increased with the decrease of the pathological grade, and the increase of lymph node metastasis and capsule invasion. Conclusion The abnormal expression of p53 and ki-67 proteins might play an important role in the pathogenesis and development of thyroid carcinoma. It has an important clinical value for early diagnosis, prognosis evaluation and proper treatment of thyroid carcinoma to determine the expression of p53 and ki-67 proteins simultaneously.