ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the antitumor efficacy of Ad-TD-RFP for human nasopharyngeal carcinoma cells (C666-1) in vitro and in vivo.</p><p><b>METHODS</b>The oncolytic effects of Ad-TD-RFP and control virus dl11520 on C666-1 cells were determined by cytotoxicity assay (MTS assay). Viral replication of Ad-TD-RFP and dl11520 was detected at different time points (24 h, 48 h, 72 h and 96 h) by tissue culture infective dose (TCID(50)) in C666-1 cells implanted subcutaneously into the flank in each of BALB/c nude mice. The xenografts were injected intratumorally with Ad-TD-RFP or dl1520 to investigate their effects on tumor growth.</p><p><b>RESULTS</b>The concentration for 50% of maximal effect (EC(50)) values of Ad-TD-RFP and dl1520 were (107.6 ± 3.2) pt/cell and (174.1 ± 4.0) pt/cell, respectively (t = 22.6, P < 0.001). The Ad-TD-RFP replication was 3-14 folds more than dl1520 replication at four time points (24 h, 48 h, 72 h and 96 h) in C666-1 cells (t values were 33.6, 23.4, 20.8 and 17.3, respectively, P < 0.001). The average tumor volumes of PBS group, dl1520 group and Ad-TD-RFP group were (1765.5 ± 713.9) mm(3), (1036.9 ± 623.8) mm(3), and (420.8 ± 238.7) mm(3), respectively (F = 12.0, P < 0.05) on day 67 after treatment.</p><p><b>CONCLUSIONS</b>The antitumour efficacy of the novel oncolytic adenovirus Ad-TD-RFP for human nasopharyngeal carcinoma C666-1 cells is superior to that of dl1520 in vitro and in vivo. The outcome of this study provides an experimental basis for the treatment of human nasopharyngeal carcinoma by viral gene therapy.</p>
Subject(s)
Animals , Female , Humans , Mice , Adenoviridae , Classification , Genetics , Carcinoma , Cell Line, Tumor , Genetic Vectors , Mice, Inbred BALB C , Mice, Nude , Nasopharyngeal Neoplasms , Therapeutics , Oncolytic Virotherapy , Oncolytic Viruses , Genetics , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To investigate the impact of all-trans retinoic acid (ATRA) on chemosensitivity to esophageal squamous cell carcinoma EC9706 cells in vitro and its mechanism.</p><p><b>METHODS</b>EC9706 cells were routinely cultured as the control group. The experimental group was divided into three groups. The ATRA group with ATRA in final concentration of 1 µmol/L; the 5-Fu group with 5-Fu in final concentration of 50 mg/L; the combined treatment group with ATRA in final concentration of 1 µmol/L and 5-Fu 50 mg/L. The cell apoptosis was detected by terminal deoxynucleotidy transferase mediated dUTP nick end labelling (TUNEL). The cell cycle and apoptosis were detected by flow cytometry.</p><p><b>RESULTS</b>The results of TUNEL showed that in the combined treatment group appeared a large number of apoptotic cells, and their nuclei were stained brown, with a positive rate of 89.7%. There was a significant difference in the comparison with the ATRA group (38.3%) and 5-Fu group (40.3%) (P < 0.05). The flow cytometry showed that the ATRA + 5-Fu group had a significantly higher apoptosis rate (76.9% ± 2.7%) than that in the ATRA group (38.2% ± 2.6%) and 5-Fu group (45.2% ± 2.3%) (P < 0.05). The ratio of cells in G(1) phase increased in the ATRA + 5-Fu group (83.4% ± 3.0%), significantly higher than (48.2% ± 2.5%) in the ATRA group and (53.2% ± 2.6%) in the 5-Fu group (P < 0.05). The ratio of cells in S + G(2)/M phase was decreased in the ATRA + 5-Fu group, with a significant difference (P < 0.05) when compared with other groups. There was no significant difference between the ATRA group and 5-Fu group (P > 0.05) in the apoptosis rate and the proportion of cells at different phases.</p><p><b>CONCLUSION</b>ATRA can induce apoptosis of esophageal carcinoma EC9706 cells in vitro. The combination of ATRA and 5-Fu may enhance the chemotherapeutic efficacy.</p>
Subject(s)
Humans , Antimetabolites, Antineoplastic , Pharmacology , Antineoplastic Agents , Pharmacology , Apoptosis , Carcinoma, Squamous Cell , Pathology , Cell Cycle , Cell Line, Tumor , Drug Synergism , Esophageal Neoplasms , Pathology , Fluorouracil , Pharmacology , Tretinoin , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanism of apoptosis of EC9706 tumor-bearing nude mice induced by all-trans retinoic acid (ATRA).</p><p><b>METHODS</b>Human esophageal carcinoma cell line EC9706 cells were inoculated into nude mice to establish the solid tumor model. The tumor models were divided into the following groups: ATRA group, fluorouracil group, the two-drugs combination group, and with an equal volume fraction of solvent as the control group. The nude mice were sacrificed after 10 days of medication. TUNEL staining was used to detect cell apoptosis. RT-PCR was used to detect the expression level of mRNA and immunohistochemistry was used to detect the expression level of protein of caspase-3 and survivin, the apoptosis-related genes in the tumor tissue.</p><p><b>RESULTS</b>The apoptosis rates of the ATRA group, 5-Fu group and ATRA + 5-Fu group were 44.3%, 39.7% and 91.0%, respectively. There was a significant difference in comparison with the control group (0.7%), and the ATRA group had no significant difference compared with that of the fluorouracil group (P > 0.05), but the apoptosis rate of the two-drugs combination group was significantly higher than that in the two single-drug groups (P < 0.05). The average gray value of caspase-3 protein expressed in the control group was 46.12 ± 0.33 and the relative expression of caspase-3 mRNA was 0.14 ± 0.03, both were significantly lower than that in the ATRA group, 5-Fu group and the two-drugs combination group (P < 0.05). The average gray value of survivin protein expressed in the control group was 96.07 ± 0.13 and the relative expression of survivin mRNA was 0.84 ± 0.04, both were significantly higher than those of other groups (P < 0.05). The ATRA group had no significant difference compared with the fluorouracil group (P > 0.05), but the two-drugs combination group was significantly different compared with the single-drug groups (P < 0.05).</p><p><b>CONCLUSION</b>Apoptosis in the EC9706 tumor cells in nude mice can be induced by ATRA. The mechanism may be related with down-regulation of the level of survivin gene expression and up-regulation of the level of caspase-3 gene expression.</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Antimetabolites, Antineoplastic , Pharmacology , Antineoplastic Agents , Pharmacology , Apoptosis , Caspase 3 , Genetics , Metabolism , Cell Line, Tumor , Esophageal Neoplasms , Metabolism , Pathology , Fluorouracil , Pharmacology , Inhibitor of Apoptosis Proteins , Genetics , Metabolism , Mice, Nude , Neoplasm Transplantation , RNA, Messenger , Metabolism , Tretinoin , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate PTEN expression and mutation status in the development of cervical adenocarcinoma.</p><p><b>METHODS</b>Immunohistochemistry study of PTEN protein was performed on 42 cases of cervical adenocarcinoma, 20 cases of cervical glandular intraepithelial neoplasia and 28 cases of normal cervix tissue samples. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to detect the presence of mutation of exons 5 and 8 of PTEN gene.</p><p><b>RESULTS</b>Positive expression rates of PTEN protein were 54.8% (23/42), 25.0% (5/20) and 100% (28/28) in cervical adenocarcinoma, cervical glandular intraepithelial neoplasia and normal cervix tissues, respectively. There were significant differences among the 3 groups (P < 0.05). Positive expression rates of PTEN protein were 47.4% (9/19), 20.0% (2/10) and 92.3% (12/13) in mucinous, endometrioid and the other variants of cervical adenocarcinoma, respectively. Mutation rates at exon 5 and exon 8 of PTEN gene were 19.0% (8/42), 45.0% (9/20) and 0 in cervical adenocarcinoma, cervical glandular intraepithelial neoplasia and normal cervix tissue, respectively. There were significant differences among 3 groups (chi(2) = 4.29, chi(2) = 12.70; P < 0.05). The mutation rates were 21.1% (4/19) and 40.0% (4/10) in mucinous and endometrioid variants of cervical adenocarcinoma, respectively. There was no mutation at exons 5 and 8 of PTEN gene detected in other variants of cervical adenocarcinoma.</p><p><b>CONCLUSION</b>The development of cervical adenocarcionomas is correlated with the mutation and absence of the protein expression of PTEN, likely in the early phase of their carcinogenesis.</p>
Subject(s)
Female , Humans , Adenocarcinoma , Genetics , Metabolism , Adenocarcinoma, Mucinous , Genetics , Metabolism , Carcinoma, Endometrioid , Genetics , Metabolism , Uterine Cervical Dysplasia , Genetics , Metabolism , Cervix Uteri , Metabolism , Exons , Mutation , PTEN Phosphohydrolase , Genetics , Metabolism , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Uterine Cervical Neoplasms , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the mRNA and protein expression of nucleostemin (NS) in human esophageal squamous cell carcinoma.</p><p><b>METHODS</b>The mRNA and protein expression of NS were detected in 31 mucosal atypical hyperplasia specimens, 62 esophageal squamous cell carcinoma specimens and the matched normal esophageal mucosa samples by RT-PCR and immunohistochemistry method, respectively.</p><p><b>RESULTS</b>The positive expression rate of NS protein in normal esophageal mucosa, atypical hyperplasia and esophageal squamous cell carcinoma was 17.7% (11/62), 41.9% (13/31) and 69.4% (43/62), respectively. There was a significant difference among the above three groups (chi2 = 33.676, P < 0.01). The expression levels of NS mRNA in esophageal squamous cell carcinoma (0.971 +/- 0.121) was significantly higher than that in the atypical hyperplasia (0.913 +/- 0.085) and also in the normal esophageal mucosa (0.866 +/- 0.103; F = 14.829, P < 0.01). The expression level of both NS protein and mRNA was positively correlated with histological grade, infiltration depth, and lymph node metastasis (P < 0.05), but not with age, gender or pathological type (P > 0.05).</p><p><b>CONCLUSION</b>Our results indicate that nucleostemin mRNA and protein are over-expressed in human esophageal squamous cell carcinoma, and it may be related with its oncogenesis.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Metabolism , Pathology , Carrier Proteins , Genetics , Esophageal Neoplasms , Metabolism , Pathology , Esophagus , Pathology , GTP-Binding Proteins , Gene Expression Regulation, Neoplastic , Hyperplasia , Lymphatic Metastasis , Mucous Membrane , Metabolism , Neoplasm Invasiveness , Neoplasm Staging , Nuclear Proteins , Genetics , Precancerous Conditions , Metabolism , Pathology , RNA, Messenger , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effects of lutein on apoptosis and its mechanism.</p><p><b>METHOD</b>The cells of human esophageal carcinoma EC9706 were grown in RPMI medium containing 10% bovine serum and were treated with lutein at 100 microg x mL(-1) concentration. Flow cytometry was employed to investigate the effects of lutein on cell apoptosis of EC9706 cells. Histochemistry was performed to determine apoptosis-related protein expresion.</p><p><b>RESULT</b>Flow cytometry analyses revealed that lutein increased EC9706 cell apoptosis ratio when treated with lutein 100 microg x mL(-1) at 96 h. Lutein decreased the expression of Bcl-2 protein and increased the expression of Bax protein in EC9706 cells.</p><p><b>CONCLUSION</b>Lutein could inhibit mitosis and stimulate apoptosis of EC9706 cells. The apoptotic effect may result from the down-regulation of expression of Bcl-2 and up-regulation expression of Bax.</p>
Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Carcinoma, Squamous Cell , Metabolism , Pathology , Cell Line, Tumor , Esophageal Neoplasms , Metabolism , Pathology , Lutein , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , bcl-2-Associated X Protein , MetabolismABSTRACT
Objective To investigate the expression and significance of death receptor 4(DR4) and DR5 in human craniopharyngioma.Methods The expression of DR4 and DR5 was determined by immunohistochemistry and in situ hybridization in 28 samples of craniopharyngioma and 25 samples of normal brain tissue.Results With low expression in partial normal brain tissue,DR was expressed highly in all of the craniopharyngioma samples.High DR expression in craniopharyngioma tissue differed from low DR expression in normal brain tissue(P0.05).Conclusions High DR expression is prevalent in craniopharyngioma tissue.This may contribute to the apoptosis-induced therapy of craniopharyngioma.The control of DR expression lays in protein level.This may contribute to the selective induced-apoptosis of tumor necrosis factor-related apoptosis-induced ligand.
ABSTRACT
Objective To investigate the expression and significance of tumor necorisis factor related apoptosis induled ligand receptor(TRAILR) in human craniopharyngioma.Methods The expression of TRAILR was determined by immunohistochemistry and in situ hybridization in 24 samples of craniopharyngioma and 16 samples of normal brain tissue.Results With low decoy receptor(DcR) expression in partial craniopharyngioma cells and low death receptor(DR) expression in partial normal brain cells,DR was expressed highly in all craniopharyngioma samples while DcR in most normal brain tissue. High DR expression and low DcR expression in craniopharyngioma tissue differed from low DR expression and high DcR expression in normal brain tissue(P