This study was aimed at investigating the effects of demethylase fat mass and obesity-associated protein(FTO)and methyltransferase methyltransferase like protein 3(METTL3),key molecules in N6-methyladenosine(m6A)modification,on the replication and proliferation of Japanese encephalitis virus(JEV).Recombinant lentiviruses were generated by packaging the FTO and green fluorescent protein into lentiviral vectors.Neuro2a cells,a mouse neuroblastoma cell line,were infected with the lentivirus,and stable FTO-expressing cell lines were obtained through puromycin selection.Successful overexpression of FTO was confirmed through fluorescence microscopy,real-time quantitative PCR,and western blot analysis.When Neuro2a cells overexpressing FTO were infected with JEV,the overexpression of FTO decreased JEV replication in the cells,and increased the expression of interferon(IFN)and related molecules.Additionally,treatment of JEV-infected Neuro2a cells with the METTL3-specific inhibitor STM2457 resulted in a dose-dependent decrease in JEV replication and viral protein expression.These findings suggested that lowering m6A methylation levels inhibits JEV replication,thus shedding light on the regulatory role of methylation modification in JEV replication.

This study was aimed at investigating the effects of demethylase fat mass and obesity-associated protein(FTO)and methyltransferase methyltransferase like protein 3(METTL3),key molecules in N6-methyladenosine(m6A)modification,on the replication and proliferation of Japanese encephalitis virus(JEV).Recombinant lentiviruses were generated by packaging the FTO and green fluorescent protein into lentiviral vectors.Neuro2a cells,a mouse neuroblastoma cell line,were infected with the lentivirus,and stable FTO-expressing cell lines were obtained through puromycin selection.Successful overexpression of FTO was confirmed through fluorescence microscopy,real-time quantitative PCR,and western blot analysis.When Neuro2a cells overexpressing FTO were infected with JEV,the overexpression of FTO decreased JEV replication in the cells,and increased the expression of interferon(IFN)and related molecules.Additionally,treatment of JEV-infected Neuro2a cells with the METTL3-specific inhibitor STM2457 resulted in a dose-dependent decrease in JEV replication and viral protein expression.These findings suggested that lowering m6A methylation levels inhibits JEV replication,thus shedding light on the regulatory role of methylation modification in JEV replication.

OBJECTIVETo study the responses of different pancreatic cancer cells to stimulations by nerve growth factor (NGF) and explore the role of Trk-A in such responses.

METHODSFive pancreatic cancer cell lines (MIA-PaCa-2, PANC-1, SW-1990, AsPC-1, and BxPC-3) were exposed to different concentrations of NGF (0, 4, 20, 100, and 500 ng/ml). MTT and Matrigel invasion method were used to observe the changes in the cell proliferation and invasion ability. Trk-A expression in these cells was detected by PCR and Western blotting, and the relations of Trk-A expression to the cell proliferative and invasive abilities following NGF treatment were analyzed.

RESULTSNGF at 100 ng/ml most obviously stimulated the cell proliferation, and PANC-1 cells showed the highest while AsPC-1 cells showed the least sensitivity to 100 ng/ml NGF stimulation. Matrigel invasion test showed that NGF enhanced the invasiveness of PANC-1 and MIA-PaCa-2 cells but produced only limited effect on AsPC-1 cells; the effect of NGF was completely inhibited by the Trk-A inhibitor CEP701. The expression levels of Trk-A mRNA and protein were the highest in PANC-1 cells and the lowest in AsPC-1 cells.

CONCLUSIONNGF can enhance the proliferation and invasiveness of pancreatic cancer cells, and this effect is possibly mediated by Trk-A protein.

Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Neoplasm Invasiveness ; Nerve Growth Factor ; pharmacology ; Pancreatic Neoplasms ; metabolism ; pathology ; Receptor, trkA ; genetics ; metabolism

OBJECTIVETo construct a plasmid carrying small interfering RNA (siRNA) targeting human C-erbB2 gene (pGenesil- erbB2) and test its effect on Her-2 expression at the post-transcriptional level in human colon cancer cell lines HT-29 cells that highly express erbB2.

METHODSA HT-29 cell line that highly expressed CerbB-2 was selected using immunohistochemical method. The double-stranded siRNA targeting human CerbB-2 cDNA and the negative control fragment were cloned into pGenesil-1 vector, and after identification and sequence analysis, the constructed pGenesil-erbB2 plasmid was transfected into the selected HT-29 cell line.

RESULTSThe pGenesil-erbB2 plasmid was successfully constructed and stably transferred into HT-29 cells. The transfection resulted in significant inhibition of Her-2 protein expression in the HT-29 cells, as shown by Western blotting.

CONCLUSIONThe pGenesil-erbB plasmid we constructed can be stably transfected into HT-29 cells to inhibit the expression of Her-2 protein, and can be useful in further studies of increasing the radiosensitivity of HT-29 cell lines.

Base Sequence ; Genes, erbB-2 ; genetics ; HT29 Cells ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Transfection

OBJECTIVE: To determine the clinical application of the new classification of adenocarcinoma of esophagogastric junction (AEG). METHODS: The data of cancer of distal esophagus, cancer of cardia, and proximal gastric cancer were reviewed. Clinicopathologic characteristics, surgical modes and survival were analyzed according to Siewert's standards. RESULTS: Among the 203 patients that were up to the standard, 29 had adenocarcinoma of the distal esophagus (Type I), 80 had true carcinoma of cardia (Type II), and 94 had subcardial carcinoma (Type III). The 5-year survival rates of the 3 types of patients after the operation were 34% for Type I, 27.5% for Type II, and 24.5% for Type III (P<0.05). Further analysis of the patients with curative resection suggested there was no significant difference in the 5-year survival rates, with 37.5% for Type I, 34.5% for Type II, and 33.3% for Type III (P>0.05). CONCLUSION Difference has been found in the clinicopathologic characteristics of the 3 types of adenocarcinoma of the esophagogastric junction. The exact relation of the 3 types is still unknown. The TNM classification, complete tumor resection and the extent of lymph node metastasis are critical for the prognosis of the patients.
Adenocarcinoma ; classification ; mortality ; surgery ; China ; Esophageal Neoplasms ; classification ; mortality ; surgery ; Esophagectomy ; Esophagogastric Junction ; pathology ; surgery ; Female ; Humans ; Male ; Retrospective Studies ; Survival Analysis ; Survival Rate

OBJECTIVETo investigate the expression of beta-protein 1 (BP(1)) gene, a novel member of DLX homeobox gene family, in lung cancer tissue and its relationship with clinical features of lung cancer.

METHODSRT-PCR was employed for detecting BP(1) gene expression in the lung cancer tissues, adjacent tissues, non-cancer lung tissues of 46 lung cancer patients.

RESULTSThirty-six lung cancer tissues and 6 adjacent tissues but none of the normal tissues were found to have BP(1) gene overexpression, showing significant difference in BP(1) expression between the tissues (P<0.01). Significant difference in BP1 gene overexpression was noted between well differentiated cancers (13 of out 21) and poorly differentiated cancers (22 of the 25), but not between cancers of different stages or between squamous carcinoma and adenocarcinoma.

CONCLUSIONBP(1) gene expression is up-regulated in human lung cancer in related to the differentiation level of lung cancer but not to the clinical stage.

Adenocarcinoma ; genetics ; pathology ; Carcinoma, Squamous Cell ; genetics ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Homeodomain Proteins ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics