摘要
目的研究抑癌基因LRIG1对膀胱癌细胞BIU87生物学特性的影响。方法用脂质体介导的基因转染技术,将pLRIG1-GFP质粒转染人膀胱癌BIU87细胞,用G418(800 mg／L)筛选出抗性克隆。Western blot检测转染前后LRIG1和EGFR蛋白水平的变化;MTT法绘制转染前后细胞的生长曲线;Boyden小室和同质黏附实验观察转染前后细胞侵袭和黏附能力的变化。结果转染pLRIG1-GFP组LRIG1蛋白表达水平较对照组和转染空载体组明显升高,而EGFR蛋白表达水平较对照组和转染空载体组显著降低;转染pLRIG1-GFP组细胞的生长速度较对照组和转染空载体组明显减慢;转染pLRIG1-GFP后的BIU87细胞侵袭和黏附能力显著降低(P<0．05)。结论 LRIG1是一种抑癌基因,通过参与形成EGFR的负反馈环路,对膀胱癌细胞BIU87的生长、侵袭和转移有显著的抑制作用。
Objective To investigate whether the expression of tumor suppressor gene LRIG1 can suppress the invasive or metastatic ability of bladder cancer cells. Methods Plasmid pLRIG1-GFP was transfected into bladder cancer cell （BIU8） by Lipofectamine2000, and cells that can express LRIG1 stably was screened out by G418 （800 mg/L）. Changes of LRIG1 and EGFR protein levels were measured by Western blot;Growth curve of the LRIG1 gene transfected bladder cancer cells was drawn with the results of MTT assay;Then Boyden chamber and cell-cell adhesion were used to detect the difference of invasion and metastases between transfected and non-transfected cells. Results LRIG1 protein level in pLRIG1-GFP transfected BIU87 cells was significantly higher than that in control BIU87 cells and pEGFP-N1 transfected BIU87 cells. EGFR protein level in pLRIG1-GFP transfected BIU87 cells was significantly lower than that in control BIU87 cells and pEGFP-N1 transfected BIU87 cells. The growth curve of the LRIG1 gene transfected ceils by MTT revealed that the LRIG1 gene transfected cells had less proliferation than control ceils and pEGFP-N1 transfected BIU87 cells. As compared with control cells and pEGFP-N1 transfected BIU87, the invasion and adhesion ability of pLRIG1-GFP transfected cells was decreased obviously （ P 〈 0.05）. Conclusion LRIG1, as a tumor suppressor gene, participates in construction of negative feedback loop of EGFR to inhibit bladder cancer cells from growth, invasion and metastasis.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第12期1543-1545,共3页
Chinese Journal of Experimental Surgery
