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目的观察正常和不同病变程度的口腔黏膜上皮细胞对α干扰素(IFN-α)敏感性的差异,为IFN-α的抗肿瘤应用和机制的研究提供基础。方法分别培养正常口腔黏膜上皮细胞株(NOK)、口腔黏膜异常增生上皮细胞株(DOK)、口腔表皮样癌细胞株(KB),取传至第3代对数生长期细胞接种于细胞培养板,对照组每孔加入2 m L含10%胎牛血清的完全培养液,实验组设IFN-α浓度100、500、1 000 U/m L 3个浓度组,实验组每孔分别加入含相应浓度IFN-α的完全培养液,常规培养24 h后检测细胞增殖能力、细胞凋亡和细胞周期变化情况。结果不同浓度的IFN-α分别作用于3种细胞,对NOK细胞株的增殖能力无影响,而DOK、KB细胞株各实验组与对照组相比增殖能力明显减慢,差异均有显著统计学意义(均P<0.01)。浓度500 U/m L IFN-α对NOK细胞株的凋亡无影响,但有促进DOK、KB细胞株凋亡的作用。浓度500 U/m L IFN-α对NOK细胞株周期无影响,DOK细胞株G0/G1期细胞比例显著减少,而G2/M期细胞比例显著增加,KB细胞株G2/M期细胞比例显著增加。结论正常上皮细胞对IFN-α不敏感,而DOK、KB细胞株对IFN-α敏感;IFN-α有抑制DOK、KB细胞株的增殖能力、促其凋亡的作用,且细胞周期阻滞于G2/M期。
Objective To observe the difference of sensitivity to interferon α (IFN-α) between normal and different pathological changes of oral mucosa epithelial cells, and to provide a basis for the anti-tumor application and mechanism of IFN-α. Methods Normal oral mucosa epithelial cell line (NOK), oral mucosa dysplasia epithelial cell line (DOK) and oral epidermoid carcinoma cell line (KB) were cultured and passaged to the 3rd generation of logarithmic growth phase cells inoculated into the cell culture plate The control group was supplemented with 2 ml of complete culture solution containing 10% fetal bovine serum. Each group in the experimental group was given 3 concentrations of IFN-α at concentrations of 100, 500 and 1 000 U / mL. Concentration of IFN-α in the complete broth, routine culture 24 h after the detection of cell proliferation, apoptosis and cell cycle changes. Results Different concentrations of IFN-α, respectively, on the three kinds of cells had no effect on the proliferation of NOK cell lines, but the proliferation ability of DOK and KB cell lines was significantly lower than that of the control group, the difference was statistically significant Significance (all P <0.01). The concentration of 500 U / m L IFN-α had no effect on the apoptosis of NOK cell lines, but had the effect of promoting the apoptosis of DOK and KB cell lines. The concentration of 500 U / m L IFN-α had no effect on the cell cycle of NOK, the percentage of cells in G0 / G1 phase decreased significantly while the proportion of cells in G2 / M phase increased significantly, while the proportion of cells in G2 / M phase increased significantly . CONCLUSION: Normal epithelial cells are not sensitive to IFN-α, while DOK and KB cell lines are sensitive to IFN-α. IFN-α can inhibit proliferation of DOK and KB cell lines and promote apoptosis. Cell cycle arrest G2 / M period.