ABSTRACT
Following targeted gene virotherapy, the suppression of tumorigenicity 13 (ST13) gene was inserted into the double-regulated oncolytic adenovirus SG500 to ensure more safety and potent antitumor activity against colorectal cancer in vitro and in vivo. We generated the ST13-expressing oncolytic adenovirus SG500-ST13, with which colorectal carcinoma cell lines SW620 and HCT116, and the lung fibroblast cell line WI38, were infected. Crystal violet staining was carried out to detect the cytopathic effect in cells, and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method was used to assay cell viability. The effect of apoptosis induced by SG500-ST13 was confirmed by Hoechst staining and the TdT-mediated dUTP-biotin nick-end labeling method. To further identify the antitumor effects of SG500-ST13 on HCT116 xenografts in Balb/c nude mice, the induction of cell death was assessed by hematoxylin-eosin staining. Immunohistochemical study was also carried out.
Subject(s)
Adenoviridae/genetics , Colorectal Neoplasms/genetics , Colorectal Neoplasms/therapy , Genetic Therapy/methods , Oncolytic Virotherapy/methods , Adenoviridae/physiology , Animals , Apoptosis/drug effects , Benzimidazoles/metabolism , Cell Death , Cell Line, Tumor , Cell Survival , Coloring Agents/metabolism , Cytopathogenic Effect, Viral/genetics , Cytopathogenic Effect, Viral/physiology , Eosine Yellowish-(YS)/metabolism , Female , Fluorescent Dyes/metabolism , Gene Expression , Genetic Vectors , Gentian Violet/metabolism , HCT116 Cells , Hematoxylin/metabolism , Humans , Immunohistochemistry , In Situ Nick-End Labeling/methods , Mice , Mice, Nude , Tetrazolium Salts/metabolism , Thiazoles/metabolism , Xenograft Model Antitumor AssaysABSTRACT
Interleukin (IL)-24 is an excellent therapeutic gene for cancer therapy. In this work, IL-24 was inserted into Ad.sp-E1A(Delta24), an oncolytic adenovirus with a 24-bp deletion in the E1A gene, which was driven by the survivin promoter to form Ad.sp-E1A(Delta24)-IL-24. Ad.sp-E1A(Delta24)-IL-24 has an excellent antitumor effect in vitro for human nasopharyngeal, liver, lung, and cervical carcinoma cell lines but does no or little damage to normal cell lines L-02 and WI38. Furthermore, it achieved nearly complete inhibition (although not elimination) of NCI-H460 lung carcinoma growth in nude mice. The antitumor efficacy of Ad.sp-E1A(Delta24)-IL-24 on NCI-H460 cells was clearly mediated by apoptosis, because it induced caspase-3 and poly(ADP-ribose) polymerase cleavage. This is the first report of Ad.sp-E1A(Delta24)-IL-24 with such an excellent, broad, and specific antitumor effect in vitro and nearly complete inhibition of lung tumor growth in vivo.
Subject(s)
Adenoviridae/genetics , Interleukins/genetics , Microtubule-Associated Proteins/genetics , Oncolytic Viruses/genetics , Promoter Regions, Genetic/genetics , Retinoblastoma Protein/genetics , Xenograft Model Antitumor Assays , Adenoviridae/physiology , Animals , Apoptosis , Cell Line, Tumor , Cell Proliferation , Cell Survival , Cytopathogenic Effect, Viral/immunology , Genetic Therapy , Humans , Inhibitor of Apoptosis Proteins , Interleukins/therapeutic use , Mice , Oncolytic Virotherapy , Oncolytic Viruses/physiology , Survivin , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>To investigate the association between hyperglycemia and outcome in elderly patients with acute ST segment elevation myocardial infarction (STEMI) underwent primary percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>This retrospective analysis was performed on 284 elderly patients (age > or = 60 years) with acute STEMI underwent primary PCI between January 2000 to April 2004 in our department. Patients were divided into 3 groups according to the level of blood glucose on admission: group A, < 7.8 mmol/L; group B, 7.8 - 10.9 mmol/L; group C, > or = 11.0 mmol/L.</p><p><b>RESULTS</b>(1) The proportion of female in group B and group C was greater than that of group A (33.3% vs. 26.5%, P < 0.01; 40.2% vs. 26.5%, P < 0.01). The hospital stay time of group B and group C was significantly longer than that of group A (16.0 days vs. 13.9 days, P < 0.05; 16.6 days vs. 13.9 days, P < 0.05). There were more patients with history of hypertension in group C than that in group A (72.1% vs. 54.9%, P < 0.01). (2) After PCI, the proportion of patients with TIMI myocardial perfusion grade (TMPG) 0-1 in group B and C was greater than that of group A (22.6% vs. 13.3%, P < 0.05; 34.1% vs. 13.3%, P < 0.05). The proportion of patients with TMPG 3 in group B and C was less than that in group A (74.3% vs. 84.4%, P < 0.05; 57.6% vs. 84.4%, P < 0.05). The complication rate of PCI was significantly higher in group C than in group A (42.5% vs. 20.6%, P < 0.01) and group B (42.5% vs. 26.6%, P < 0.01). IABP use was significantly more in group C than that in group A (19.5% vs. 4.9%, P < 0.01) and group B (19.5% vs. 6.4%, P < 0.01). (3) There were more patients with grade of Killip class > or = 2 in group C than that in group A (44.8% vs. 23.5%, P < 0.01) and group B (44.8% vs. 27.7%, P < 0.01). The in-hospital mortality rate (8.0% vs. 1.1%, P < 0.05) and one-year mortality rate (18.7% vs. 3.4%, P < 0.05) of group C were significantly higher than those in group A.</p><p><b>CONCLUSION</b>Hyperglycemia at admission was associated with poor tissue perfusion, cardiac function and prognosis in elderly patients with acute STEMI underwent primary PCI.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Angioplasty, Balloon, Coronary , Blood Glucose , Hyperglycemia , Myocardial Infarction , Blood , Therapeutics , Prognosis , Retrospective Studies , Treatment OutcomeABSTRACT
AIM: The aim of the present study was to further improve the therapeutic effects for human hepatocellular carcinoma (HCC) and reduce the damage in normal cells using a novel chemo-gene-virotherapeutic strategy. METHODS: An oncolytic adenoviral vector (ZD55) similar to the typical oncolytic adenovirus ONYX-015, with a deletion of E1B-55K gene, was employed to express the second mitochondria-derived activator of caspases (Smac) protein by constructing a recombinant virus ZD55-Smac. The enhanced cytotoxicity of the combined treatment of ZD55-Smac with cisplatin or 5-fluorouracil (5-FU) was evaluated in several HCC cell lines. Moreover, the negative effects on normal cells have been tested in human normal liver cell lines L-02 and QSG-7701 cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and apoptotic cell staining. RESULTS: According to our observation, ZD55-Smac is superior to ONYX-015 in sensitizing chemotherapy, ZD55-Smac used in conjunction with chemotherapy was found to exhibit obviously enhanced cytotoxicity in HCC cells, yet significantly abolished the negative toxicity in normal cells by utilizing the tumor selective replication vector and reducing the dosage. CONCLUSION: This chemo-gene-virotherapeutic (cisplatin or 5-FU+ZD55-Smac) strategy is superior to the conventional chemo-gene or chemo-viro approach.
Subject(s)
Adenoviridae/genetics , Antineoplastic Agents/therapeutic use , Carcinoma, Hepatocellular/drug therapy , Liver Neoplasms/drug therapy , Oncolytic Virotherapy , Base Sequence , Carcinoma, Hepatocellular/therapy , Cell Line, Tumor , DNA Primers , Humans , Liver Neoplasms/therapyABSTRACT
<p><b>BACKGROUND</b>Hyperglycemia has been shown to be a powerful predictor of poor outcome after ST-segment elevation myocardial infarction (STEMI). This study aimed to evaluate the effect of admission glucose on microvascular flow after successful primary percutaneous coronary intervention (PCI) in patients with STEMI.</p><p><b>METHODS</b>Successful primary PCI was performed in 267 patients with STEMI. The maximum ST elevation of single electrocardiogram (ECG) lead before and 60 minutes after PCI was measured, and patients were then divided into 3 groups according to the degree of ST-segment resolution (STR): absent (<30%), partial (30% to 70%) or complete (> or =70%).</p><p><b>RESULTS</b>Of the 267 patients, 48 (18.0%) had absent STR, 137 (51.3%) experienced partial STR, and 82 (30.7%) had complete STR. The degree of STR decreased with increasing admission glucose levels (P=0.032), and patients with hyperglycemia (serum glucose level > or =11 mmol/L) were more likely to have absent STR (P=0.001). Moreover,hyperglycemia was an independent predictor of incomplete STR (odds ratio, 1.870; 95% confidence interval, 1.038 to 3.371, P=0.037).</p><p><b>CONCLUSIONS</b>Hyperglycemia on admission is associated with abnormal coronary microvascular reperfusion in patients with STEMI after successful primary PCI, which may contribute, at least in part, to the poor outcomes in these patients.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Angioplasty, Balloon, Coronary , Methods , Electrocardiography , Glucose , Metabolism , Hyperglycemia , Blood , Pathology , Myocardial Infarction , Blood , Therapeutics , Odds RatioABSTRACT
<p><b>AIM</b>To investigate the cellular signal transduction pathway of vascular smooth muscle cell (VSMC) proliferation stimulated by insulin-like growth factor-1 (IGF-1).</p><p><b>METHODS</b>Rabbit aortic VSMCs was cultured in 3 groups. Cell proliferating ability was determined by measuring cell number and mitochondrial dehydrogenase (MD) activity (MTT assay). Wortmannin (WT), the specific inhibitor of phosphatidylinositol 3-kinase (PI3K), was used to evaluate indirectly the possible involvement of PI3K. Western blotting was used to detect the protein expression of phosphatase PTEN. Diphosphate action of PTEN on its specific substrate diC16PIP3 was measured by green reagent method.</p><p><b>RESULTS</b>IGF-1 (100 microg/L) increased cell number and MD activity by 2.8-3.8 fold. WT markedly inhibited VSMC proliferation and completely abolished the above effects of IGF-1. IGF-1 inhibited PTEN activity in a concentration-(10-100 microg/L) and time--(3 min-24 h) dependent manner (P < 0.01).</p><p><b>CONCLUSION</b>IGF-1 increases VSM proliferation by increasing PI3K activity and inhibiting PTEN activity.</p>
