Effects Of TIN2 Mutation On Telomereand Chromosome Of Human Gastric Epithelial Cell Line GES-1

[Objective] TIN2 is a telomere protective protein that plays an important role in the telomere protein complex.Studies have recently shown that structural changes in the telomere-protected protein complex,Shelterin,can cause telomere function disorders,leading to a range of diseases.This study based on the establishment of a stable TIN2 gene mutation eukaryotic vector and empty vector of human gastric mucosal epithelial cells GES-1 to investigate the effect of TIN2 gene mutation on telomere and chromosome of GES-1 human gastric mucosal epithelial cells.[Methods] The human gastric mucosal epithelial cells GES-1 of TIN2 gene mutation eukaryotic vector and empty vector was established stablely.Immunofluorescence/telomere peptide nucleic acid fluorescence in situ hybridization(PNA-Fish)assay to detect whether TIN2 gene mutation can cause damage to telomeric DNA and trigger telomere dysfunction lesion(TIF);Chromosome experiments to see if TIN2 gene mutation leads to chromosomal aberrations(ring chromosome,chromosome end-end fusion);Southern blot analysis showed whether the TIN2 gene mutation had an effect on the relative length of telomeres.[Res?lts] 1.The human gastric mucosal epithelial cells GES-1 with stable TIN2 gene mutation eukaryotic vector and empty vector were successf?lly constructed.2.The PNA-Fish experiment found that there were no obvious lesions in the empty vector group and the blank group.More than three telomere dysfunction lesions appeared in the GES-1 cell group with TIN2 gene mutation eukaryotic vector,and the difference was statistically significant.(P < 0.05).3.Chromosomal analysis revealed that GES-1 cell chromosomal aberrations were observed in the GES-1 cell group of the TIN2 gene-mutant eukaryotic vector,whereas no chromosomal aberrations were observed in the empty vector group and the blank group.The difference was statistically significant(P < 0.05).4.Southern blot analysis showed that there was no significant change in the relative length of telomeres in the empty and blank groups.The relative length of telomeres in GES-1 cell group with TIN2 gene mutation eukaryotic vector became shorter.The difference was statistically significant(P < 0.05).[Conclusions] In human gastric mucosal epithelial GES-1 cells,TIN2 gene mutation res?lted in telomere dysfunction lesions and shortened mean telomere length,as well as chromosomal aberrations.It indicates that TIN2 gene mutation may play a certain role in the occurrence of gastric cancer.