重组质粒载体pCMV-LRIG2转染BIU-87细胞株的效应研究

目的研究携带有LRIG2基因的重组质粒载体pCMV-LRIG2对膀胱癌细胞株BIU-87生物学行为的影响。方法用脂质体介导的基因转染技术，将携带有LRIG2基因的重组质粒载体pCMV-LRIG2转染至膀胱癌细胞株BIU-87，用G418筛选出抗性克隆后，Western blot检测转染前后LRIG2和EGFR蛋白表达水平变化，MTT法绘制转染前后细胞生长曲线，Transwell法检测转染前后细胞侵袭力，同质黏附实验观察转染前后细胞黏附能力。结果转染pCMV-LRIG2组LRIG2蛋白表达水平显著高于末转染组和空载体转染组；转染pCMV-LRIG2组细胞生长速度较未转染组和空载体转染组明显减慢，侵袭力显著低于未转染组和空载体转染组，而黏附能力显著高于未转染组和空载体转染组。结论LRIG2有类似于抑癌基因的作用，通过对EFG-EGFR信号转导途径发挥负反馈抑制作用，从而抑制BIU87细胞的生长、侵袭和转移。

The research on the effect of BIU-87 cell transfected by recombined plasmid pCMVLRIG2

Objective To investigate the effects of recombined plasmid pCMV-LRIG2 carrying LRIG2 gene on the biological behaviors of human bladder cancer BIU-87. Methods The recombined plasmid pCMV-LRIG2 was transfected into BIU-87 cell by lipofectamine 2000, and G418 was used to screen out the positive clone. Then Western-blot was used to measure the expression of LRIG2 and EGFR protein, MTT assay was used to drawn out the growth curve of the cell, and transwell method and cell-cell adhesion were used to detect the invasion and metastasis ability among transfected cell, non-transfected cell, and pCMV-transfected cell. Results Among the LRIG2-transfected group, non-transfected group and pCMV- transfected group, the expression of LRIG2 protein was significantly higher in the LRIG2-transfected group than in the other two groups, and the expression of EGFR in LRIG2-transfected group was significantly lower. The growth curve of the cells by MTT showed that the LRIG2-transfected group had less proliferation than the other two groups. Contrast to non-transfected group and pCMV-group, the invasion ability of transfected group decreased significantly. However, the adhesion ability of transfected group increased significantly. Conclusion The LRIG2 gene may act as a tumor suppressor gene by participating in the construction of negative feedback loop of EGFR, so as to inhibit bladder cancer cells from growth, invasion and metastasis.