新基因hepaCAM对膀胱癌细胞体外生物学影响

The effect of hepaCAM gene on biological behavior of TCCB cell in vitro

目的:研究肝细胞粘附分子(Hepatocyte cell adhesion molecule,hepaCAM)对人类膀胱癌T24细胞体外生物学影响。方法:脂质体法转染pEGFP-N2-hepaCAM质粒入T24细胞,激光共聚焦显微镜检测其蛋白定位;筛选稳定表达细胞株;Western blot检测蛋白表达。细胞粘附、基质胶侵袭实验及MTT实验研究该基因对细胞粘附力、活动力及增殖力的影响。结果:hepaCAM在单个细胞中分布在细胞核周边区域;相互连接细胞中主要分布于细胞间相互接触区域。获得稳定表达hepaCAM基因的T24细胞。体外粘附实验、基质胶侵袭实验证明实验组细胞粘附力及活动力明显高于空白组及阴性对照组。MTT法检测转染hepaCAM基因细胞增殖力明显低于空白组及阴性对照组。结论:hepaCAM基因可显著抑制人类膀胱癌T24细胞恶性行为,可能是通过增强细胞-基质间粘附及抑制细胞增殖力从而影响肿瘤细胞的浸润和转移。

Objective：To investigate the effect of transfection with wild hepaCAM gene on biological behavior of the transitional cell carcinoma of bladder in vitro.Methods：Human TCCB cell line T24 was transfected with pEGFP-N2-hepaCAM,mock plasmid pEGFP-N2 with lipofectamine.Laser confocal microscopy was used to detect the location of hepaCAM in cells.Western bolt was used to determine the expression of target of hepaCAM in stable cells.The cell adhesive and motility ability were tested by cell spreading assay and matrigel invasion assay.Cell proliferation ability was investigated by MTT assay.Results：In well spread cells,hepaCAM localized on the perinuclear membrane,whereas in the cells which contacts,hepaCAM predominantly accumulated at the sites of cell-cell contacts on cell membrane.Cell clone with stable expression of hepaCAM,was acquired.In vitro experiments as cell spreading assays and matrigel invasion assays showed the cell adhesion and cell motility properties of hepaCAM group were apparently enhanced compared with the non-transfection or mock-vehicle groups.The MTT assay showed cell proliferation ability in the hepaCAM group was notably decreased when compared with the non-transfection or mock-vehicle groups.Conclusion：The hepaCAM gene can restrain malignant phenotypes of the human TCCB cells in vitro,and may inhibit the TCCB invasion and metastasis by influencing the function of some adherence and proliferation factors.