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目的研究抗角蛋白单克隆抗体(mAb)对体外培养的角质形成细胞增殖活性与细胞周期的影响。方法以无血清培养基体外原代培养角质形成细胞。应用3H-TdR掺入DNA合成测定法,观察抗角蛋白16mAb(anti-K16mAb)对角质形成细胞代谢增殖的影响。用流式细胞仪分析anti-K16mAb作用后角质形成细胞的细胞周期变化。结果mAb作用后,角质形成细胞样品的cpm降低,并呈抗体剂量依赖方式。流式细胞仪分析表明,处于S期的细胞比例由27%上升至42.2%,同时处于G1期与G2期的细胞比例分别相应下降。结论anti-K16mAb可抑制体外培养的角质形成细胞的增殖活性,且将细胞阻滞于S期,无法进入G2期与M期分裂增殖。
Objective To investigate the effect of anti-keratin monoclonal antibody (mAb) on the proliferation and cell cycle of keratinocytes cultured in vitro. Methods Primary cultured keratinocytes were cultured in serum-free medium. 3H-TdR incorporation DNA synthesis assay was used to observe the effect of anti-K16mAb on the keratinocyte proliferation. Cell cycle changes of keratinocytes after anti-K16 mAb treatment were analyzed by flow cytometry. Results The cpm of keratinocyte samples decreased after mAb treatment in an antibody dose-dependent manner. Flow cytometry analysis showed that the proportion of cells in S phase increased from 27% to 42.2%, while the proportion of cells in G1 phase and G2 phase decreased correspondingly. Conclusion Anti-K16 mAb can inhibit the proliferation activity of cultured keratinocytes in vitro and arrest the cells in S phase, which can not enter the G2 phase and M phase of the proliferation.