正常和病变的口腔黏膜上皮细胞对α干扰素的敏感性差异研究

Sensitivity difference of oral epithelial cells in alpha interferon between normal and lesion

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 sensitivity differences of proliferation ability beween three different oral cells after incubation with interferon alpha（IFN-α）, and provide the basis on anti-tumor application and mechanism of IFN-α. Methods The normal oral mucosa epithelial cell line（NOK）, oral dysplasia epithelial dell line（DOK） and oral epidermoid carcinoma cell line （KB） were cultured respectively and the 3rd generations were seeded on cell culture plates. Control group was pulsed with 2 mL 10 % fetal bovine serum（FBS） complete medium in per wells. The experimental groups were incubated with 100 U/mL, 500 U/mL and 1 000 U/mL IFN respectively, which were tested after 24-hour regular culture, and the cell proliferation ability, apoptosis and cycle changing were observed. Results The different concentrations of IFN-α performed 3 kinds of cells, there was no effect on cell proliferation ability of NOK cell line, while compared with control group, DOK and KB cell lines were slowly down and the difference was statistical significance （P 〈 0.01）. The concentration of 500 U/mL IFN-α was no apoptosis affected on NOK cell lines, but DOK and KB cell lines apoptosis were promoted. The concentration of 500 U/mL IFN-α was no affected on NOK cells cycle, but G0/G1 phase DOK cell lines significantly reduced, G2/M phase cells increased significantly, and the proportion of G2/M phase KB cell lines increased significantly. Conclusion It is demonstrated that normal epithelial cells are non-sensitive in IFN-α, but much high sensitivity of IFN-α in DOK and KB cell lines. The IFN-α could inhabit cells proliferation and promote apoptosis in DOK and KB cell lines, and the cell cycle arrest in G2/M phase.