ABSTRACT
AIM: To determine the expression and function of epithelial membrane protein 1 (EMP1) in colorectal carcinoma. METHODS: Colorectal samples were taken from cancer lesions and adjacent normal tissue in colorectal cancer patients immediately after endoscopic biopsy. A portion of the sample was either fixed in 4% paraformaldehyde and embedded in paraffin for immunohistochemistry or stored in liquid nitrogen for Western blot. In order to determine protein expression of EMP1 in colorectal cancer (n = 63) and normal tissue (n = 31), semi-quantitative immunohistochemistry and Western blot were utilized. For in vitro studies, the human colorectal cancer cell line SW-480 was maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum. Recombinant lentivirus mediated overexpression of EMP1 in SW-480 cells was quantified by real-time reverse transcription-polymerase chain reaction and Western blot. Control SW-480 cells were transfected with an empty vector. To further study the effect of EMP1 overexpression in SW-480 cells, cell proliferation, apoptosis, migration and invasion assays were conducted. RESULTS: Expression of EMP1 was significantly lower in colorectal cancer tissue than in normal tissue using both immunohistochemistry (39.7% vs 90.3% of tissues, P < 0.05) and Western blot (0.126 ± 0.022 vs 0.632 ± 0.053, P < 0.05). The level of EMP1 protein expression was not correlated with gender, age, or tumor location. Decreased expression of EMP1 was significantly correlated with T stage, lymph node metastasis, clinic stage, and histological grade in patients with colorectal cancer (P < 0.05). According to Kaplan-Meier analysis, low EMP1 expression correlated significantly with poor overall five-year survival (34.2% vs 64.0% survival, P < 0.05). SW-480 cells transfected with EMP1 had a lower survival fraction, higher cell apoptosis (12.1% ± 1.3% vs 3.1% ± 0.6%, P < 0.05), a significant decrease in migration and invasion (124.0 ± 17.0 and 87.0 ± 12.0, respectively vs 213.0 ± 29.0 and 178.0 ± 21.0, respectively, P < 0.05), higher caspase-9 (0.635 ± 0.063 vs 0.315 ± 0.032, P < 0.05), and lower VEGFC protein expression (0.229 ± 0.021 vs 0.519 ± 0.055, P < 0.05) relative to cells not transfected with EMP1. CONCLUSION: Low EMP1 expression in colorectal cancer is associated with increased disease severity, suggesting that EMP1 may be a negative regulator of colorectal cancer.
Subject(s)
Colorectal Neoplasms/metabolism , Neoplasm Proteins/metabolism , Receptors, Cell Surface/metabolism , Adult , Aged , Apoptosis , Biopsy , Caspase 9/metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cell Survival , Endoscopy , Female , Humans , Immunohistochemistry , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Invasiveness , Prognosis , Vascular Endothelial Growth Factor C/metabolismABSTRACT
OBJECTIVE: To observe the duration of enterovirus-71 (EV71) and coxsackievirus A 16 (CoxA16) viral shedding in stool samples of children with hand, foot and mouth disease (HFMD) infected with EV71 and CoxA16 and to explore the relationship between the duration of intestinal virus shedding and the severity of illness of children with HFMD. METHOD: Totally 113 laboratory-confirmed cases of children with HFMD infected with EV71 and CoxA16 were followed up. The stool samples were collected with the interval of 4 to7 days and the viral nucleic acids were detected by fluorescent PCR until the stool viral nucleic acids of infected children turned to be negative. The cases in EV71 group were further divided into "ordinary EV71 group" and "severe EV71 group" according to the severity of the illness. The positive rates of viral nucleic acid and the differences of distribution among different groups were analyzed by Kaplan-Meier survival analysis during the follow-up period. RESULT: The 113 cases of infected children were grouped as follows: 65 cases of EV71 positive children, 44 cases of CoxA16 positive children, 4 cases of EV71/CoxA16 mixed infection. The median duration of the stool viral nucleic acids turning to negative was 26 (18.25-32.50) days in EV71 group and 27 (14.50-33.75) days in CoxA16 group (Z = 1.51, P > 0.05). At 1, 4, 6 and 10 weeks, the positive rates of stool viral nucleic acid of children with HFMD in EV71 group were 100%, 48.1%, 17.2% and 0 respectively. At 1, 4 and 6 weeks, the positive rates of stool viral nucleic acid of children with HFMD in CoxA16 group were 95.5%, 53.8% and 0 respectively (χ(2) = 0.18, P > 0.05). At 1, 4 and 6 weeks, the positive rates of stool viral nucleic acid of children with HFMD in ordinary EV71 group were 100%, 23.5% and 0 respectively, while at 1, 4, 6 and 10 weeks, the positive rates of stool viral nucleic acid of children with HFMD in severe EV71 group were 100%, 62.4%, 26.0% and 0 respectively (χ(2) = 5.689, P < 0.05). CONCLUSION: The duration of enterovirus shedding in stool samples of children with HFMD lasted for a long period. The maximum duration of EV71 and CoxA16 in stool of children with HFMD was 10 weeks and 6 weeks, respectively. The duration of intestinal virus shedding of children with HFMD infected with EV71 was related with the severity of the illness.
Subject(s)
Enterovirus A, Human/isolation & purification , Feces/virology , Hand, Foot and Mouth Disease/virology , Nucleic Acids/isolation & purification , Virus Shedding , Child , Child, Preschool , China/epidemiology , Coxsackievirus Infections/diagnosis , Coxsackievirus Infections/epidemiology , Enterovirus/genetics , Enterovirus/isolation & purification , Enterovirus A, Human/genetics , Female , Hand, Foot and Mouth Disease/epidemiology , Hand, Foot and Mouth Disease/prevention & control , Humans , Infant , Male , Polymerase Chain Reaction , RNA, Viral/geneticsABSTRACT
OBJECTIVE: To investigate the molecular-epidemiologic characteristics and genotypes of human calicivirus (HuCVs) in acute diarrhea patients in Hangzhou from 2009 to 2010. METHODS: Epidemiologic data and fecal specimens were collected from patients with acute diarrhea. HuCVs of 920 specimens were detected by PCR. PCR products of several positive samples were randomly selected and sequenced. All the sequences were analyzed, phylogenetically. RESULTS: 201 HuCVs positive cases were identified from 920 facal specimens (21.8%). 25 isolates would include norovirus GI-type, GII-type for 170 strains and sapovirus for 11 strains. Norovirus GI-type and GII-type were detected in four specimens at the same time. Other specimens were mixed infection with norovirus GII-type and sapovirus. Genotypes of HuCVs showed that norovirus GI subtypes were GI-1 (3 strains) and GI-2 (1 strain). Norovirus GII subtypes were GII-4/2006b variant strains (7 strains), GII-2 (1 strain), GII-7 (1 strain) and GII-4/2008 variant strains (2 strains); Sapovirus subtypes were GI-2 (5 strains), GI-1 (4 strains) and GII-1 (1 strain). The prevalence rates of HuCVs were different in seasons and age groups. CONCLUSION: HuCVs were one of the major pathogens causing acute diarrhea. Both multiple viruses and genotypes of HuCVs were found in the specimens. GII-4/2006b variant and similar strains were identified, probably as the prevalent strains from 2009 to 2010 in Hangzhou, Zhejiang province.
