1.Effects of H_2O_2 and PTEN on activation of Akt by peptide growth factors
Liu YANG ; Yanying HUO ; Yingchun HU
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
0.05).Conclusion The peptide growth factors can activate the Akt kinase by phosphorylation.This process depends on the production of H2O2 and the presence of PTEN.
2.Differential proteomic analysis of Pten~(L/L)MEFs and Pten~(△/△)MEFs cell lines
Ruifeng DUAN ; Gang LI ; Yingchun HU ; Dechang WU ; Yanying HUO
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(03):-
Objective To study protein variation between PtenL/LMEFs and Pten△/△MEFs cells.Methods Two-dimensional electrophoresis(2-DE) was employed to compare the differential expression proteins between PtenL/LMEFs and Pten△/△MEFs cells.Six differential expression proteins were digested in gel by enzyme and the mass of generated peptides was measured by matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS).The data obtained from peptide mass fingerprinting(PMF) were searched using the Internet available database and five proteins were identified.Results Compared with that of PtenL/LMEFs,expression level of proteins including phosphoglycerate mutase 1(PGAM1) and peptidyl-prolyl cis-trans isomerase C(PPIC) was up-regulated,whereas expression level of Transgelin 2 was down-regulated in Pten△/△MEFs cells.B and F proteins were both identified to be peroxiredoxin-6.They had similar molecular weight but different PI which might be caused by post-translation modification.B protein was only expressed in Pten△/△MEFs cells.Conclusion The protein profile of Pten△/△MEFs cells displayed obvious difference compared to that of PtenL/LMEFs cells.The results implied that various distinct different proteins might lead to cancer.
3.Construction of recombinant adenovirus vector expressing EGFP gene specifically in AFP producing liver cancer cells
Yujun SHI ; Changan LIU ; Jianping GONG ; Xuhong LI ; Yong PENG ; Ying MEI ; Can MI ; Yanying HUO
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To construct a recombinant adenovirus vector carrying AFP promoter to specifically express a targeting gene in hepatocellular carcinoma HepG2 cells. METHODS: Based on the Adeno-X~TM Expression system, the CMV promoter was replaced by a 300 bp a-fetoprotein promoter. The EGFP(enhanced green fluorescent protein) gene as a report gene was inserted to the multiple-cloning site(MCS). The normal liver LO2 cells, hepatocellular carcinoma HepG2 cells and HeLa cells were infected by the recombinant adenovirus, respectively. Northern blotting and fluorescence microscope were used to detect the transcription level of EGFP gene and its protein expression, respectively. RESULTS: Northern blotting showed that the target gene was markedly transcribed in HepG2 cells, but slightly in LO2 and HeLa cells. Under the fluorescence microscope, strong EGFP expression was seen in HepG2 cells but very weakly in HeLa and LO2 cells. CONCLUSION: Under the control of the 300 bp human AFP promoter, the target gene carried by the recombinant adenovirus was expressed in the AFP-producing HepG2 cells at a very high level, but not or very weakly in AFP negative cells. This adenovirus system can be used as a new, potent and specific approach for the gene-targeting therapy for the AFP producing primary hepatoma. [
4.Changes of gene methylation profile in malignant transformation of immortalized human bronchial epithelial cell line induced by alpha-particle irradiation
Yingchun HU ; Zhongmin CHEN ; Qiaodan ZHOU ; Boqiang SONG ; Gang LI ; Dechang WU ; Yanying HUO
Chinese Journal of Radiological Medicine and Protection 2011;31(4):420-424,432
Objective To identify the changes of DNA methylation profile in the process of malignant transformation of BEP2D cell induced by α particles.Methods The genomic DNAs were isolated from the malignant transformation BERP35T4 cells and immortalized human bronchial epithelial cell line BEP2D.Genomic DNAs were digested by MseI and ligated of PCR linkers.Methylated DNAs were digested by BstUI and amplified by PCR.The methylated DNA probes were prepared by labeling with Cy3 and Cy5 fluorescence dyes individually and hybridized to the methylation CpG-Island microarray.The hybridization results were scanned and analyzed.Intensity values were quality controlled and normalized.The normalized data were used to identify the differentially expressed genes based on a 1.5 fold difference of the expression level.Results There were 16 genes which showed changes of methylation level in malignant transformation BERP35T4 cells, 9 of them were hypermethylation and 7 were hypomethylation.These genes were including the SKIP gene, PPP3CC gene, MAP2K6 gene, KIR2DL1 gene, KIR2DL4 gene, KIR3DP1 gene, ZNF493 gene, ZNF100 gene, NKX2-5 gene, TFAP2D gene, DR1 gene, KCNJ16 gene, CCDC18 gene, FNBP1L gene, IRX4 gene, EPB41L3 gene, TCP10 gene and so on.Conclusions The DNA methylation might have effects on ionizing radiation drived tumorigenesis.
5.Development of young talented scientists by excellent young scientists fund : Review of excellent young scientists fund in the department of health sciences of National Natural Science of Foundation of China
Dou DOU ; Yuhua PENG ; Minghe HUO ; Yanying XU ; Ruijuan SUN ; Erdan DONG
Chinese Journal of Medical Science Research Management 2019;32(1):29-33
Objective The implementation and prospect of excellent young scientists fund in the department of health sciences of national natural science of foundation of china were reviewed and discussed to propose suggestions for future development.Methods The applications and grants of excellent young scientists fund in health sciences field from 2012 to 2017 wereanalyzed among different medical branches,regions and supporting institutions.The age,gender,title and academic rank of both applicants and awardees were analyzed.Results The applications and grants of excellent young scientists fund vary greatly in the different branches of medical science,as well as in the different regions and supporting institutions.The person in charge of such projects has been better developed after being awarded.Conclusions The establishment and implementation of the excellent young scientists fund has promoted the growth of young medical scientists.