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目的通过比较不同硬度基底对肝细胞和肝癌细胞迁移特征的影响,研究肿瘤细胞迁移行为变化的成因。方法运用免疫荧光染色、形态学分析法和Transwell侵袭实验,观察不同硬度基底上HCCLM3和L02细胞的形态特征及对细胞运动能力进行检测和定量分析。结果 (1)相对于极软(0.5 kPa)和极硬(玻璃)基底,HCCLM3和L02细胞在较软基底(4 kPa)上具有较高的迁移速率和迁移净距离,且L02细胞表现出高的迁移效率。(2)HC-CLM3和L02细胞在不同硬度基底上均方位移趋势一致,较软基底上L02细胞具有较高的方向持续能力。(3)0.5和1 mg/mL三维胶原基质中,HCCLM3细胞穿透基底膜的个数分别显著多于L02细胞穿透基底膜的个数;加入40μg/mL水解酶抑制剂GM6001后,HCCLM3细胞穿透基底膜的个数显著增加,而L02细胞穿透基底膜的个数显著减少。结论 (1)二维较软基底上,L02细胞因有较高的方向持续能力而表现出高的迁移效率。(2)三维胶原基质中,HCCLM3细胞以不同迁移模式适应周围环境,从而表现出更大的侵袭能力。
OBJECTIVE: To study the genesis of migration behavior of tumor cells by comparing the effect of different hardness substrates on the migration characteristics of hepatocytes and hepatoma cells. Methods Immunofluorescence staining, morphological analysis and Transwell invasion assay were used to observe morphological characteristics and cell motility of HCCLM3 and L02 cells on different hardness substrates. Results (1) HCCLM3 and L02 cells had higher migration rates and net migration distances on softer substrates (4 kPa) relative to very soft (0.5 kPa) and very hard (glass) substrates, and L02 cells showed high The migration efficiency. (2) The tendency of HC-CLM3 and L02 cells to move in the same direction on different hardness substrates was consistent, and LO2 cells on the soft substrate had a higher directional sustainability. (3) In the three-dimensional collagen matrix of 0.5 and 1 mg / mL, the number of HCCLM3 cells penetrating the basement membrane was significantly higher than that of L02 cells penetrating the basement membrane respectively. After addition of 40μg / mL hydrolase inhibitor GM6001, HCCLM3 cells The number of penetrating basement membrane significantly increased, while the number of L02 cells penetrating basement membrane significantly reduced. Conclusion (1) Two-dimensional soft base, L02 cells due to the higher direction of sustained capacity and showed high migration efficiency. (2) Three-dimensional collagen matrix, HCCLM3 cells in different migration patterns to adapt to the surrounding environment, thus showing greater invasive ability.