Knockdown of IGF2BP2 inhibits colorectal cancer cell proliferation, migration and promotes tumor immunity by down-regulating MYC expression / 细胞与分子免疫学杂志

Objective To investigate the effect of insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) on the proliferation, migration and tumor immune microenvironment of colorectal cancer cells and its possible molecular mechanism. Methods The Cancer Genome Atlas (TCGA) database was used to analyze the expression levels of IGF2BP2 and MYC in colorectal cancer and adjacent tissues. The expression of IGF2BP2 in HCT-116 and SW480 human colorectal cancer cells was silenced by RNA interference (RNAi), and the silencing effect was detected by quantitative real-time PCR. After knocking down IGF2BP2, colony formation assay, CCK-8 assay and 5-ethynyl-2'-deoxyuridine (EdU) assay were employed to detect cell colony formation and proliferation ability. TranswellTM assay was used to detect cell migration ability. Quantitative real-time PCR was used to detect the mRNA expression of IGF2BP2, MYC, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β) and interleukin-10 (IL-10). The protein expression of IGF2BP2 and MYC was detected by western blot. The binding ability of IGF2BP2 and MYC in HCT-116 cells was detected by quantitative real-time PCR after RNA immunoprecipitation. Results The results of TCGA database showed that the expression of IGF2BP2 and MYC in colorectal cancer tissues was significantly higher than that in adjacent tissues, and the survival time of colorectal cancer patients with high expression of IGF2BP2 was shorter. After silencing IGF2BP2, the viability, proliferation and migration of HCT-116 and SW480 cells were decreased. The mRNA expression of MYC, TGF-β and IL-10 in IGF2BP2 knockdown group was significantly decreased, while the expression of TNF-α mRNA was increased. The expression of MYC protein and the stability of MYC mRNA were significantly decreased. RIP-qPCR results showed that IGF2BP2 could bind to MYC mRNA. Conclusion Knockdown of IGF2BP2 inhibits colorectal cancer cell proliferation, migration and promotes tumor immunity by down-regulating MYC expression.

Effect of VEGF165b on biological characteristics of human hepatocellular carcinoma HepG2 cells / 吉林大学学报(医学版)

Objective:To study the effect of vascular endothelial growth factor 165b (VEGF165b) on the biological characteristics of human hepatocellular carcinoma HepG2 cells,and to explore its mechanisr.Methods;The HepG2 cells were divided into blank group (treated with transfection reagent),control group (transfected with pcDNA3.0 expression vector) and pcDNA-VEGF165b group (transfected with pcNDA-VEGF165b).MTT assay was used to detect the survival rate of HepG2 cells;RT-PCR and Western blotting method were used to detect the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG2cells;Transwell chamber assay was used to measure the migration ability of HepG2 cells.Results:Compared with blank group,there was no significant changes in the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG2 cells in control group (P＞0.05).Compared with blank group,the expression levels of VEGF165b mRNA and protein in the HepG2 cells in PcDNA-VEGF165b group were significantly increased (P＜0.05),and the expression levels of VEGF165 mRNA and protein were significantly decreased (P＜0.05).There was no significant difference in the survival rates between blank group and control group (P＞0.05).The survival rate of HepG2 cells in PcDNA-VEGF165b group was lower than those in blank group and control group,but the differences were not statistically significant (P＞0.05).The cell migration experiment results showed that there was no significant difference in the migration rate between blank group and control group (P＜0.05).Compared with blank group and control group,the migration rate of HepG2 cells in PcDNA-VEGF165b group was significantly reduced (P＜ 0.05).Conclusion:VEGF165b can inhibit the expressions of VEGF165 mRNA and protein,and VEGF165b has no effect on the proliferation of hepatocellular carcinoma cells;but it can reduce the migration of hepatocellular carcinoma cells.

A Winged-Helix Transcription Factor Foxg1 Induces Expression of Mss4 Gene in Rat Hippocampal Progenitor Cells

Foxg1 (previously named BF1) is a winged-helix transcription factor with restricted expression pattern in the telencephalic neuroepithelium of the neural tube and in the anterior half of the developing optic vesicle. Previous studies have shown that the targeted disruption of the Foxg1 gene leads to hypoplasia of the cerebral hemispheres with severe defect in the structures of the ventral telencephalon. To further investigate the molecular mechanisms by which Foxg1 plays essential roles during brain development, we have adopted a strategy to isolate genes whose expression changes immediately after introduction of Foxg1 in cultured neural precursor cell line, HiB5. Here, we report that seventeen genes were isolated by ordered differential displays that are up-regulated by over-expression of Foxg1, in cultured neuronal precursor cells. By nucleotide sequence comparison to known genes in the GeneBank database, we find that nine of these clones represent novel genes whose DNA sequences have not been reported. The results suggest that these genes are closely related to developmental regulation of Foxg1.

The study of changes on NKT cells of experimental autoimmune encephalomyelitis (EAE) mice / 细胞与分子免疫学杂志

AIM: To observe the changes of the number of NKT cells in spleens and livers of induced model of experimental autoimmune encephalomyelitis (EAE), and to study the role NKT cells play in the immunoregulation of EAE. METHODS: C57BL/6 mice were immunized with MOG peptide and received clinical evaluation daily. The mice were sacrificed at the fastigium and the splenic and hepatic lymphocytes were isolated. The changes of NKT cells in normal and EAE C57BL/6 mice were detected by flow cytometry. RESULTS: The percent of NKT cells in lymphocytes of different organs of EAE model were greater decreased than in that of normal mice. The percent of NKT cells in splenic lymphocytes of normal mice was 2.22± 0.14, while that in EAE mice was 1.94±0.07 (P < 0.05). The percent of NKI cells in hepatic lymphocytes of normal mice was 5.52±2.17, while that in EAE mice was 2.67± 1.41 (P < 0.05). CONCLUSION: The proliferation of splenic and hepatic NKT cells in C57BL/6 mice are inhibited in EAE model, which may indicate that the immune function conducted by NKT cell is down regulated in EAE mice.