ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects and the mechanisms of cell growth inhibition in hepatocellular carcinoma cells after induction with antisense survivin-liposome (LIP) complex, and to provide evidence in treatment for hepatocellular carcinoma and tumors expressing survivin.</p><p><b>METHODS</b>Survivin ODNs was transfected into HepG2 cells mediated by LiP reagent. The expression of survivin mRNA and protein was detected by RT-PCR and Western blot. MTT assay was applied to determine cell proliferation in HepG2 cells. Active caspase-3 and apoptosis rate were evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Cell cycle was analyzed by flow cytometry in the cell cycle-synchronized hepatocellular carcinoma cells treated with the antisense compound.</p><p><b>RESULTS</b>Antisense compound efficiently down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC(50) of 250 nmol/L. Its maximal effect was achieved at a concentration of 600 nmol/L, when expression levels were down-regulated by 80%, as revealed by gradually increase of caspase-3-like protease activity and apoptosis rate in a time-dependent manner. Morphological apoptotic changes such as membrane blebbing, loss of microvilli, cytoplasmic vasculization, condensation of cytoplasm and nucleus, chromatin fragmentation, and apoptosis and cell growth inhibition were observed. In the cell cycle-synchronized hepatocellular carcinoma cells, antisense compound induced cell cycle arrest followed by apoptosis. After treated with low concentration of compound, the cell cycle was arrested at S phase or G2/M phase; while at high concentration, the cell cycle was mainly arrested at S phase. Apoptosis was obviously observed and the rate of apoptosis was increased in a time and concentration-dependent manner.</p><p><b>CONCLUSION</b>Antisense survivin has significant inhibitory effect on growth of hepatocellular carcinoma cells in vitro. This is associated with cell cycle arrest and apoptosis.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Hepatocellular , Pathology , Caspase 3 , Caspases , Metabolism , Cell Cycle , Cell Proliferation , Inhibitor of Apoptosis Proteins , Liposomes , Liver Neoplasms , Pathology , Microtubule-Associated Proteins , Metabolism , Pharmacology , Neoplasm Proteins , Metabolism , Pharmacology , Oligonucleotides, Antisense , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To study the expression and associations of three survivin splicing variants in gastric cancer and normal gastric mucosa, and to evaluate the prognostic significance.</p><p><b>METHODS</b>Real time quantitative RT-PCR was used to detect the expression of three survivin splicing variants in tumor and matched normal gastric mucosa specimens from 77 cases with gastric cancer.</p><p><b>RESULTS</b>The expression of three survivin splicing variants than upregulated significantly in gastric cancer than those in normal mucosas (P< 0.01). In cancer tissues, the expression rates of survivin, survivin-2B, survivin-deltaEx3 were 100%, 79.8% (61/77), 64.9% (50/77) respectively. The survival rate was significantly lower in the patients with high survivin expression than those with low survivin expression (P< 0.01).</p><p><b>CONCLUSION</b>Among three survivin splicing variants, the expression level of wild-type survivin mRNA is an important predictor for prognosis.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Biomarkers , Follow-Up Studies , Gastric Mucosa , Pathology , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , Neoplasm Staging , Prognosis , Protein Isoforms , Genetics , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms , Genetics , Pathology , Transcription, GeneticABSTRACT
Objective To study the sources and the relationship between the management and the outcome of hemorrhage after cephalic pancreatoduodenectomy.Methods The clinical data of 370 patients who underwent pancreatic resection at the Lihuili Hospital and the Second Affiliated Hospital of Zhejiang University were retrospectively analyzed.Results Postoperative bleeding occurred in 35 patients with 11 deaths.Among those intraabominal bleeding occurred in 14 cases and gastrointestinal hemorrhage occurred in 22,with one case suffering from both.Bleediug developing within 72 hours after operation in 12 cases (early-stage group),which was caused by improper intraoperative homeostasis.In other 23 cases,bleeding 72 hours after operation(later stage group)was caused by the erosion following pancreatic and/or bile leakage.Relaparotomy was performed in 13 cases and endoscopic homeostasis was performed in 3. Relaparotomy or endoscopic homeostasis was superior to that of conservative therapy in the early-stage group (P0.05).Pancreatic or bile leakage was identified as the significant risk factors for the postoperative bleeding.Conclusions In order to prevent the postoperative hemorrhage and to reduce the mortality of pancreatic resection,skillful techniques,expeditious homeostasis,proper management of stump pancreas and the prevention of pancreatic and bile leakage are essential.