中文摘要：

目的通过比较融合生长肝细胞（L02）与肝癌细胞（HCCLM3）对不同硬度基底（0．5、4kPa和玻璃）的响应，探查两类细胞融合生长差异的成因。方法运用实时显微摄影技术、免疫荧光染色、流式细胞技术及WesternBlot—ring等试验方法分别检测融合生长ID2和HCCLM3细胞在不同硬度基底上的形态特征、骨架构象、E—cad和Integrinβ1表达分布，以及Src激酶活性水平的变化。结果（1）融合生长L02细胞呈圆形或立方形，HCCLM3细胞呈多角形，铺展和极化更为明显；随基底硬度增加，L02细胞圆度随时间变化幅度较小，HCCLM3细胞变化幅度较大。（2）融合生长的L02细胞皮质下呈现环形骨架，E—cad定位于细胞一细胞接触处，HCCLM3中皮质骨架环不完整，胞内骨架沿基底呈放射状分布，E—cad呈弥散分布于细胞质中。（3）随基底硬度增加，L02和HCCLM3细胞中E—cad表达显著降低（P〈0．01），而p-Src和Integrinβl的表达量则显著增加（P〈0．01），HCCLM3较L02细胞变化明显。结论皮质下环形骨架装配与E—cad在细胞一细胞定位呈正相关，基底硬度对肝癌细胞钙黏素与整合素黏附系统平衡调节的影响较对肝细胞的影响明显。

英文摘要：

Objective To investigate the cause of differences in confluent growth between hepatic（L02） and hep atoma carcinoma（HCCLM3） cells by comparing responses of the two cells to different substrate stiffness （0.5, 4 kPa and glass）. Methods The realtime photomicrography, immunofluorescence staining, flow cytometry, and Western Blotting techniques were respectively employed to observe the morphological characteristics, the cy toskeleton conformation and the distribution of Ecad of confluent L02 and HCCLM3 cells on different substrates, and test the changes in expression of Ecad, IntegrinlM and pSrc. Results （ 1 ） Confluent L02 cells displayed a round or cubic shape, while HCCLM3 cells showed a polygon shape. The morphology of HCCLM3 cells were spread and polarized more obviously than that of L02 cells. With the increase of substrate stiffness, the variation of L02 cells with time was smaller than that of HCCLM3 cells. （2） The cytoskeleton of confluent L02 cells showed a ringlike conformation under the cortex, and Ecad was located at the cellcell contact sites. However, the ring like cytoskeleton of HCCLM3 cells was incomplete and distributed radially along the basement, while Ecad was dispersed in cytoplasm. （3） As the substrate stiffness increased, expression of Ecadherin in both L02 and HC CLM3 cells was significantly decreased （P 〈 0.0l）, while the level of pSrc and integrinβl was increased signifi cantly, with greater changes in HCCLM3 cells than in L02 cells. Conclusions The assembling of cortical ringlike cytoskeleton was positively correlated with the location of Ecad at the cellcell contact sites. The substrate stiff ness showed a more obvious impact on the balanced regulation between cadherin and integrin mediated adhesion system of hepatocarcinoma cells than that of hepatic cells.