hepaCAM基因对膀胱癌细胞体外生物学的影响

Effect of hepaCAM gene on biological behavior of TCCB cells in vitro

目的研究野生型hepaCAM基因对人类膀胱癌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。细胞黏附、基质胶侵袭实验及MTT实验研究该基因对细胞黏附力、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。体外黏附实验、基质胶侵袭实验证明实验组细胞黏附力及活动力明显高于空白组及阴性对照组(P<0.01)。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(P<0.01)。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-基质间黏附及抑制细胞增殖力,从而对抗肿瘤细胞的浸润和转移。

Objective To investigate the effect of transfection of wild hepatocyte cell adhesion molecule （hepaCAM） on the biological behaviors of the T24 cells of transitional cell carcinoma of bladder （TCCB）. Methods Human TCCB cell line T24 was transfected with pEGFP-N2-hepaCAM, mock plasmid pEGFP-N2 with lipofectamine. Laser confocal microscopy was used to observe the location of hepaCAM in cells. Western blotting was used to determine the expression of hepaCAM in stably transfected cells. The cell adhesive and mo- tility ability were tested by cell spreading assay and matrigel invasion assay. Cell proliferation ability was inves- tigated by MTT assay. Results In well spread cells, hepaCAM was localized on the perinuclear membrane, whereas in the cells contacting together, it predominantly accumulated at the sites of cell-cell contacts on cell membrane. Cell clones with stable expression of hepaCAM were acquired. Cell spreading assay and matrigel invasion assay showed the cell adhesion and cell motility properties of hepaCAM transfected cells were apparently enhanced compared those with no transfection or mock-vector cells. MTT assay showed cell proliferation ability in the hepaCAM group was notably decreased when compared with the non-transfection or mock-vector groups. Conclusion hepaCAM 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.