幽门螺旋杆菌上胃癌细胞中p16的表达.pdf

Helicobacterpylori(Hp) upregulatesthe expression ofp16 in gastriccancercells Abstract Objective: The mechanism of Helicobacter pylori pathogenicity is not well understood, although bacterial virulence factor has been associated with the development of gastric carcinogenesis. p16 is a very important facter of carcinoma,and now overexpression of p16 protein has been found in gastric carcinoma.So, it is necessary to study the expression of p16 in gastric cancer and relationshipwithHelicobacterpylori infection.Theaimofthis studywas to analyze whetherHelicobacterpylori infection inducesp16 expression andthe mechanism in humangastriccancercellsSGC-7901. Methods:TheSGC-7901cellswereco-incubatedwithHelicobacterpylori atvarious concentrations for different time. Changes in the morphology of SGC-7901cells ① wereobservedbyinversionmicroscope. Expressionofp16proteinwasassessedby ② Western-blot. ExpressionofP16mRNAwasassessedbyreversetranscription-PCR. ③ Plasmid construction of fragments of P16 promoter(900bp,358bp,155bp) Cells ④ ⑤ weretransientlytransfectedwiththep16-lucvector. Aluciferaseassaywasusedto ⑥ monitoractivationoftheP16genepromoter. Results: After SGC-7901 cells infected with Helicobacterpylori, under the light ① microcopy,there were frank vacuolation in the cytoplasm around thenucleus. ②infection of SGC-7901gastric epithelial cellswithHelicobacterpylori resulted in an increase in p16 protein,and Helicobacter pylori (MOI=100:1) significantly increased the expression of p16 protein in SGC-7901 cells in 24h post-treatment. ③CoculturewithHelicobacterpylori significantlyenhancedthesteady-statelevelsof P16mRNAinSGC-7901cells. P16promoterwashighlyactivedinSGC-7901cells ④