MDCK细胞成瘤相关蛋白的筛选及验证

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中图分类号：Q942.6 文献标识码：A 文章编号： 1000-4440（2025）10-2009-09

Abstract：In order to clarify the tumorigenic mechanism of Madin-Darby caninekidney（MDCK）cells， based on data-independent acquisition mass spectrometry （DIA-MS）technology，non-tumorigenic MDCK cells and highlytumorigenic MDCK cellswere used as test materials inthisstudytoanalyze theproteinexpression differencesof different tumorigenic MDCK cells. The results showed that a totalof11359 proteinswereidentifiedin non-tumorigenic MDCKcellsand highly tumorigenic MDCK cells，of which thenumber of differentially expressed proteinswith fold change （FC）>1.5 and corrected P<0.05 was 163. In non-tumorigenic MDCKcels，93diferentiallyexpressedproteinswereup-regulatedand7Odiffrentiallyexpressedproteinswere down-regulated.Diferentiallyexpressedproteinsweremainlyinvolvedinproteinphosphorylation，intracelularsignal transduction and negativeregulation of peptidaseactivity，andtheirfunctions wereenriched in extracelular space.Results of real-time fluorescencequantitativePCRand Westernblotanalysisof twokeytumorigenic proteins，doublecortin-likekinase1 （DCLKI）andtissue factorpathwayinhibitor2（TFPI2）showed thattherelative expressionlevelofDCLK1 genein non-tumorigenic MDCKcellswashighlysignificantlylowerthanthatintumorigenic MDCKcells，and therelativeexpression levelof TFPI2 genewas highlysignificantlyhigher thanthatintumorigenicMDCKcells.Theresultsof thisstudyprovide potential molecular regulatory targets forthe tumorigenic mechanism of MDCK celsand the screening of tumor markers.

KeyWords：MDCKcell；data-independent acquisition mass spectrometry（DIA-MS）；tumorigenic；doublecortinlikekinase1（DCLKl）；tissue factor pathway inhibitor2（TFPI2）

MDCK细胞系是由Madin和Darby于1958年从美国雌性可卡犬的肾脏中提取的上皮样细胞而建立的，通常以贴壁方式生长。（剩余14845字）