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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 study the biological effects of wild-type hepaCAM on human bladder cancer T24 cells in vitro. Methods The plasmid pEGFP-N2-hepaCAM was transfected into T24 cells by lipofectamine. The protein localization was determined by laser confocal microscopy. The stable cell lines were selected and the protein expression was detected by Western blot. Cell adhesion, matrigel invasion assay and MTT assay were used to study the effect of the gene on cell adhesion, activity and proliferation. Results HepaCAM was distributed in the periphery of the nucleus in a single cell, and intercellular connected cells were mainly distributed in the intercellular contact area. T24 cells stably expressing the hepaCAM gene were obtained. In vitro adhesion experiment and matrigel invasion experiment demonstrated that the cell adhesion and activity in experimental group were significantly higher than those in blank group and negative control group (P <0.01). MTT assay hepaCAM gene transfected cell proliferation was significantly lower than the blank group and the negative control group (P <0.01). Conclusion The hepaCAM gene can significantly inhibit the malignant behavior of T24 human bladder cancer cells, which may be through the enhancement of cell-matrix adhesion and inhibition of cell proliferation, thereby preventing tumor cell invasion and metastasis.