1.Abnormal O-glycosylation mediated by the deficiency of molecular chaperone Cosmc or T-synthase regulates the expression of miRNAs in colorectal cancer exosomes
Tianbo GAO ; Yang GE ; Guangyu AN ; Jiannan YAO ; Yuliang JIANG ; Heshu LIU ; Rui YAN
Journal of Capital Medical University 2025;46(3):401-409
Objective To explore the effects of aberrant O-glycosylation modifications induced by the knockout of Cosmc or T-synthase genes on the expression profiles of miRNAs in exosomes derived from colorectal cancer cells and to reveal the molecular mechanisms of O-glycosylation in the development of colorectal cancer and identify potential biomarkers for early diagnosis and treatment.Methods This research specifically targets the Cosmc or T-synthase genes in the human colorectal cancer cell line HCT116 to create stable cell lines exhibiting abnormal O-glycosylation with CRISPR/Cas-9 gene editing technology.Exosomes originating from these colorectal cancer cells were isolated and authenticated.A microarray chip equipped with primer sequences for 16 miRNAs closely associated with colorectal cancer was employed to assess the differential expression of miRNAs within these exosomes with fluorescent quantitative polymerase chain reaction(PCR).And then,a cohort of miRNAs that exhibited significant and consistent changes in expression levels across the exosomes from both cell lines was selected.These miRNAs were further validated independently with traditional fluorescent quantitative PCR.Subsequently,data from The Cancer Genome Atlas Program(TCGA)database containing patient information on colorectal cancer was harnessed.Employing R programming language,Gene Set Enrichment Analysis(GSEA)was conducted on the upregulated miRNA to investigate the downstream pathways significantly impacted and the malignant biological behaviors they may influence.Results The absence of either Cosmc or T-synthase genes results in the dysregulation of O-glycosylation in colorectal cancer cells,leading to the exposure of Tn antigens.This,in turn,affects the expression levels of specific miRNAs in exosomes derived from these cells.Specifically,the expression of hsa-miR-125b-1-3p was downregulated,while that of hsa-miR-218-5p was upregulated.Notably,hsa-miR-218-5p were found to be closely associated with the epithelial-mesenchymal transition(EMT)process in tumor cells,which is a key mechanism in cancer progression.Conclusion It elucidates that the aberrant O-glycosylation mediated by the knockout of Cosmc or T-synthase genes significantly influences the expression of certain miRNAs in exosomes from colorectal cancer cells,potentially affect the EMT process in colorectal cancer and thereby promoting distant metastasis.Given the inherent stability and detectability advantages of colorectal cancer-derived exosomes,the altered expression levels of miRNAs within these exosomes may serve as indicators of the stated of abnormal O-glycosylation in colorectal cancer.These findings suggest that exosomal miRNAs have potential as biomarkers for monitoring disease progression and therapeutic efficacy.Consequently,this could pave the way for more personalized diagnostic and treatment strategies tailored to individual colorectal cancer patients,enhancing the precision and effectiveness of clinical management.
2.Abnormal O-glycosylation mediated by the deficiency of molecular chaperone Cosmc or T-synthase regulates the expression of miRNAs in colorectal cancer exosomes
Tianbo GAO ; Yang GE ; Guangyu AN ; Jiannan YAO ; Yuliang JIANG ; Heshu LIU ; Rui YAN
Journal of Capital Medical University 2025;46(3):401-409
Objective To explore the effects of aberrant O-glycosylation modifications induced by the knockout of Cosmc or T-synthase genes on the expression profiles of miRNAs in exosomes derived from colorectal cancer cells and to reveal the molecular mechanisms of O-glycosylation in the development of colorectal cancer and identify potential biomarkers for early diagnosis and treatment.Methods This research specifically targets the Cosmc or T-synthase genes in the human colorectal cancer cell line HCT116 to create stable cell lines exhibiting abnormal O-glycosylation with CRISPR/Cas-9 gene editing technology.Exosomes originating from these colorectal cancer cells were isolated and authenticated.A microarray chip equipped with primer sequences for 16 miRNAs closely associated with colorectal cancer was employed to assess the differential expression of miRNAs within these exosomes with fluorescent quantitative polymerase chain reaction(PCR).And then,a cohort of miRNAs that exhibited significant and consistent changes in expression levels across the exosomes from both cell lines was selected.These miRNAs were further validated independently with traditional fluorescent quantitative PCR.Subsequently,data from The Cancer Genome Atlas Program(TCGA)database containing patient information on colorectal cancer was harnessed.Employing R programming language,Gene Set Enrichment Analysis(GSEA)was conducted on the upregulated miRNA to investigate the downstream pathways significantly impacted and the malignant biological behaviors they may influence.Results The absence of either Cosmc or T-synthase genes results in the dysregulation of O-glycosylation in colorectal cancer cells,leading to the exposure of Tn antigens.This,in turn,affects the expression levels of specific miRNAs in exosomes derived from these cells.Specifically,the expression of hsa-miR-125b-1-3p was downregulated,while that of hsa-miR-218-5p was upregulated.Notably,hsa-miR-218-5p were found to be closely associated with the epithelial-mesenchymal transition(EMT)process in tumor cells,which is a key mechanism in cancer progression.Conclusion It elucidates that the aberrant O-glycosylation mediated by the knockout of Cosmc or T-synthase genes significantly influences the expression of certain miRNAs in exosomes from colorectal cancer cells,potentially affect the EMT process in colorectal cancer and thereby promoting distant metastasis.Given the inherent stability and detectability advantages of colorectal cancer-derived exosomes,the altered expression levels of miRNAs within these exosomes may serve as indicators of the stated of abnormal O-glycosylation in colorectal cancer.These findings suggest that exosomal miRNAs have potential as biomarkers for monitoring disease progression and therapeutic efficacy.Consequently,this could pave the way for more personalized diagnostic and treatment strategies tailored to individual colorectal cancer patients,enhancing the precision and effectiveness of clinical management.
3.USING STR POLYMORPHISM TO STUDY THE GENETIC RELATIONSHIPS AMONG A CHINESE KOREAN POPULATION AND OTHER GROUPS
Ya GAO ; Tianbo JIN ; Shengbin LI
Journal of Pharmaceutical Analysis 2005;17(1):53-56
Objective To discuss the genetic relationship between Chaoxian and the other populations with STR polymorphisms, and study the origination of Chaoxian with historical data, analyze the difference among them. Methods Samples were obtained from 91 unrelated individuals of Jilin province. PCR amplification was performed using PE9700, the results were analyzed with the GeneScan software and then using the Genetic Analyzer ABI377 and Genotyper2.5. Tests for Hardy Weinberg equilibrium (HWE) and the clustering map were carried out using SPSS11.5. The DA (genetic distance) according to Nei's was calculated and a phylogenetic tree based on the neighbor-joining method using Mega2 software package was constructed. Results 81 alleles and 196 genotypes were observed, with the corresponding frequencies ranging from 0.0055 to 0.4615 and 0.0110 to 0.2747, the observed and expected of genotypes were evaluated using χ2-test and all of the loci were in accordance with Hardy Weinberg equilibrium (P>0.05). Chaoxian was clustered with the other Chinese ethnic groups, then the American Black and the White; it was clustered with the Baishan Han, the Beijing Han of China first, then the South Korean. Conclusion The differentiation among races is larger than that among minorities, and, the differentiation among minorities is related to and is consistent with their geographic location.
4.REPORT OF HLA DISTRIBUTION IN 2315 VOLUNTEER DONORS OF CHINESE BONE MARROW BANK FROM NORTHWEST CHINA
Tianbo JIN ; Ya GAO ; Hongbo ZHANG ; Junhai ZHAO ; Jianghua LAI ; Shuping LAI ; Shengbin LI
Journal of Pharmaceutical Analysis 2005;17(1):45-48
Objective To report the HLA data of volunteer donors of Chinese bank from Northwest China and characterize the distribution of HLA genes in Northwest China. Methods HLA-A, B antigens of 2315 volunteer donors were examined by the method of microlymphocytetoxicity (MLT) test .The antigen frequencies(AF) were assessed by directly counting; and based on that gene frequencies(GF) were calculated. HLA data from other population were collected and distribution characteristics were compared. With the raw data, Hardy-Weinberg equilibrium, statistical parameters of forensic medicine interest for HLA were computed. Results A total of 18 specific antigens were detected in HLA-A and the most frequent antigen was A2 . AF and GF were 0.5136 and 0.3026, respectively. A total of 42 specific antigens were detected in HLA-B and the most frequent antigen was A13. Its AF and GF were 0.1978 and 0.1044, respectively. The heterozygosity(H), polymorphism information content(PIC), discrimination power(DP) and probability of paternity exclusion (PPE) of HLA-A were 0.8215, 0.8212, 0.9356 and 0.7798 accordingly; while H,PIC, DP and PPE of HLA-B were 0.9322, 0.9322, 0.9878 and 0.9528. Conclusion The polymorphism of HLA-A,B genes is characteristic in Chinese. In this research, the genetic trait of HLA in 2315 volunteers is consistent with Northern Han population.
5.Genetic relationships of 15 populations of Guangxi Province
Tianbo JIN ; Ya GAO ; Teng CHEN ; Chunxia YAN ; Shengbin LI
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To study the genetic relatio ns hips of 15 groups of Guangxi Province, China by using STR polymorphisms. Methods We collected the blood samples of unrelated individuals from different groups and s equenced the STR data with ABI377, and then analyzed all these data with statist ical methods. Results The study on 6 STR loci showed that Jing and Miao n ationalities of Guangxi Province were close to each other, while Yao, Gelao and Mulam nationalities of Guangxi were far from the other groups. Conclusi on The 15 different ethnic groups of Guangxi, China have gene commun ications.

Result Analysis
Print
Save
E-mail