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目的探讨LKB1在Peutz-Jeghers综合征(PJS)错构瘤及肠上皮细胞中对上皮间质转化(EMT)调控作用。方法采用免疫组化法检测20例PJS错构瘤标本及10例正常肠道组织中LKB1、ecadherin、vimentin蛋白的表达。Masson三色染色法分析胶原纤维沉积。选取NCM460人肠上皮细胞株,慢病毒转染后建立LKB1敲低及空载体稳定表达株。以CCK8实验和transwell迁移实验评价LKB1敲低对细胞增殖、迁移的影响。运用WB、q PCR、免疫荧光检测LKB1敲低后对细胞中EMT相关蛋白ecadherin、vimentin、ncadherin、snail、slug表达的影响。结果与正常肠组织相比,PJS错构瘤中LKB1和ecadherin蛋白表达下降,而vimentin蛋白表达增加。Masson染色提示错构瘤中胶原纤维沉积增多,沉积面积扩大。与空载体组相比,LKB1敲低的NCM460细胞增殖、迁移能力明显增强(P<0.01),细胞中ecadherin表达降低,ncadherin、vimentin、snail、slug表达增高。结论LKB1可能通过调控EMT从而影响PJS错构瘤的形态改变及纤维化进程,提示EMT可能成为PJS治疗的新靶点。
Objective To investigate the regulatory effect of LKB1 on epithelial-mesenchymal transition (EMT) in Peutz-Jeghers syndrome (PJS) hamartomas and intestinal epithelial cells. Methods The expressions of LKB1, ecadherin and vimentin in 20 PJS hamartoma specimens and 10 normal intestinal tissues were detected by immunohistochemistry. Masson trichrome staining for collagen deposition. Select NCM460 human intestinal epithelial cell line, lentivirus transfection LKB1 knockdown and empty vector stable expression strain. The effects of LKB1 knockdown on cell proliferation and migration were evaluated by CCK8 assay and transwell migration assay. The effects of LKB1 knockdown on the expression of EMT-related proteins ecadherin, vimentin, ncadherin, snail and slug were detected by WB, q PCR and immunofluorescence. Results Compared with normal intestinal tissue, the expression of LKB1 and ecadherin in PJS hamartoma decreased and the expression of vimentin increased. The results of Masson staining showed that the deposition of collagen fibers increased and the deposition area increased. Compared with the blank vector group, LKB1-knockdown NCM460 cells had significantly enhanced proliferation and migration ability (P <0.01), decreased expression of ecadherin, and increased ncadherin, vimentin, snail and slug expression. Conclusion LKB1 may influence the morphological changes and fibrosis of PJS by regulating EMT, suggesting that EMT may be a new target of PJS treatment.