外源性CCN1促进辐射损伤L929细胞的生长和迁移

Exogenous CCN1 promotes proliferation and migration of radiation-injured L929 cells

目的观察外源性CCN1是否促进辐射损伤L929细胞生长和迁移,以探讨CCN1在放创复合伤修复中的作用。方法以4Gyγ射线辐射L929细胞作为辐射损伤细胞模型,细胞辐射后分别加入2mlCCN1或RFP条件培养液,37℃,5%CO2培养,作为CCN1组和RFP组;空白条件培养液培养辐射损伤L929细胞作为空白组。各组进行MTT试验、平板克隆形成试验、细胞周期分析和划痕愈合试验。结果与RFP组和空白组相比,CCN1条件培养液处理后,与空白组辐射损伤L929细胞增殖明显增强(P<0.05),与RFP组和空白组相比,克隆形成率明显增高[CCN1组(34.4±3.6)%,RFP组(24.5±2.9)%,空白组(29.5±3.5)%](P<0.05),培养5d时CCN1组S期细胞[(37.3±2.3)%]比RFP组[(25.2±1.9)%]和空白组[(26.9±1.3)%]增多(P<0.01),细胞迁移能力明显增强划痕愈合试验在1～5d的愈合宽度/原宽度值与RFP组和空白组相比差异显著(P<0.01)。结论外源性CCN1可促进辐射损伤的成纤维细胞生长和迁移,提示CCN1是放创复合伤修复的促进因素之一。

Objective To study the role of Cysteine-rich 61 （ Cyr61/CNN1） in repair of combined injury by observing whether exogenous CCN1 promotes the proliferation and migration of radiation-injured L929 cells. Methods A radiation model of L929 cells was induced by γ ray at a dose of 4 Gy. The irradiated L929 cells were cultured in a medium containing 2 ml adenovirus plasmids of CCN1 or RFP at 37 ℃ in an atmosphere containing 5% CO2,which served as a CCN1 group and a RFP group,respectively. Irradiated L929 cells cultured in a blank control medium served as a blank control group. Effects of CCN1 on proliferation and migration of radiation-injured L929 cells were detected by MTT assay,plate colony formation assay,cell cycle analysis,and scratch test of wound healing. Results The proliferation and colony formation rates of L929 cells cultured in a medium containing CCN1 were significantly higher than in RFP and control groups [colony formation rate：（ 34. 4 ±3. 6） % vs （ 24. 5 ±2. 9） % and （ 29. 5 ±3. 5） %,P 0. 05]. Five hours after culture,the amount of L929 cells in the S phase was larger in CCN1 group than in RFP and control groups （ 37. 3% ± 2. 3% vs 25. 2% ± 1. 9% and 26. 9% ± 1. 3% ,P 0. 01）. Scratch test of wound healing showed that the width/thick-ness ratio of migrated L929 cells in healed wounds was significantly higher in CCN1 group than in RFP and control groups （ P 0. 01）. Conclusion Exogenous CCN1 promotes the proliferation and migration of radiation-injured L929 cells,suggesting that CCN1 is one of the stimulating factors for the repair of combined injury.