肝癌细胞在Ⅳ型胶原裱衬表面的粘附特性

Adhesion of hepatocellular carcinoma cells to collagen coated surfaces.

目的定量描述肝癌细胞和肝细胞在ＩＶ型胶原裱衬表面的粘附特性，探讨细胞粘附力与ＩＶ型胶原裱衬浓度及ＩＶ型胶原与层粘连蛋白（ＬＮ）复合裱衬时ＬＮ复合浓度的相关关系。方法采用分步裱衬的方法制作有关裱衬表面，即依次在玻璃表面裱衬多聚赖氨酸和ＩＶ型胶原／ＬＮ，最后以牛血清白蛋白作用阻断非特异性粘附。细胞粘附力采用微管吸吮技术进行测定。结果细胞粘附力表现出对ＩＶ型胶原裱衬浓度较好的依赖性。当裱衬质量浓度从１μｇ／ｍｌ增加至５μｇ／ｍｌ时，肝癌细胞粘附力增加幅度［２０２×１０－１０牛顿（Ｎ），２６％］远远小于肝细胞粘附力增加幅度（１５６６×１０－１０Ｎ，７４２％），并呈现饱和效应，Ⅳ型胶原裱衬质量浓度超过２μｇ／ｍｌ时，肝癌细胞粘附力不再增加。ＬＮ与ＩＶ型胶原复合裱衬并不一定导致肝细胞和肝癌细胞在Ⅳ型胶原裱衬表面的粘附力增加。在特定的复合裱衬质量浓度（０．６２５μｇ／ｍｌ或１．２５μｇ／ｍｌ），ＬＮ使肝细胞和肝癌细胞在Ⅳ型胶原裱衬表面的粘附力达到最大值，其余浓度的ＬＮ与Ⅳ型胶原复合裱衬均引起细胞粘附力的下降。结论上述方法是定量研究细胞与胶原蛋白粘附特性的有效手段。肝癌细胞与Ⅳ型胶原的粘附特性较之肝细胞发生了明显的变化。Ｌ?

Objectives To quantitatively describe the adhesion of hepatocellular carcinoma (HCC) cells and hepatocytes to collagen /laminin coated surfaces and to investigate the relevance of cell adhesion forces to the coating concentration of collagen and, when laminin was cocoated along with collagen , the incorporation concentration of laminin. Methods A stepwise coating protocol was adopted to prepare the collagen /laminin coated surfaces, in which polyDlysine and collagen /laminin were successively coated onto glass surfaces and finally, bovine serum albumin was used to block the nonspecific adhesion. Cell adhesion forces were measured by means of micropipette aspiration technique. Results Cell adhesion forces showed good dependence on the coating concentration of collagen . However, the amplitude for the increase of adhesion forces of HCC cells (20210-10N,26%) was far more less than that for hepatocytes(1 56610-10N,742%) when coating concentration increased from 1 g/ml to 5 g/ml. No further increase in the adhesion forces of HCC cells was observed when the coating concentration of collagen IV increased beyond 2g/ml. The incorporation of laminin along with 2 g/ml collagen did not readily result in an increase in the adhesion forces of the two kinds of cells to the surfaces. Among the concentration of laminin tested, only in certain concentration(0.625 g/ml or1.25 g/ml) did the incorporation of laminin along with 2 g/ml collagen IV lead the adhesion forces of hepatocytes and HCC cells to the coated surfaces to reach their corresponding maximum values, otherwise this incorporation led the adhesion forces to decrease. Conclusion The present method is efficient in investigating cellmatrix adhesion. The cell adhesion behavior to collagen for HCC differs obviously from those for normal hepatocytes. Laminin might be an important factor for the regulation of cell adhesion forces to matrix components