Maspin基因甲基化对口腔黏膜鳞癌细胞的调控作用

Involvement of Maspin methylation in the proliferation of oral squamous cell carcinoma

目的:分析口腔黏膜鳞癌细胞增殖过程中Maspin基因甲基化的调控作用机制,为相关研究及临床治疗提供借鉴。方法:鳞状细胞癌HIOEC-B(a)P细胞系随机分为4组:5-氮-2-脱氧胞苷+曲古抑菌素A(ADC+TSA)阴性组、低(0.1μmol/L+0.05μmol/L)、中(1μmol/L+0.5μmol/L)和高剂量组(10μmol/L+5μmol/L)。以正常口腔黏膜上皮细胞作为对照组。实时定量PCR检测各组细胞Maspin基因的甲基化程度,MTT法检测各组细胞的增殖情况,4',6-二脒基-2-苯基吲哚(DAPI)染色检测细胞凋亡。采用SPSS20.0软件包对数据进行单因素方差分析。结果:与对照组相比,癌细胞的Maspin基因甲基化程度显著增强(P<0.01),与阴性组相比,中剂量和高剂量组Maspin基因甲基化程度显著减弱(P<0.05,P<0.01),低、中、高剂量组的细胞增殖抑制率显著高于阴性组(P<0.05,P<0.01)。随着药物剂量的提高,细胞凋亡程度逐渐增加(P<0.05)。结论:Maspin基因甲基化可能参与口腔黏膜鳞癌细胞的增殖过程。

PURPOSE： Explore the involvement of Maspin methylation in the proliferation of oral squamous cell carcinoma. METHODS： HIOEC-B（a）P cell line was divided into 4 groups： negative control, ADC＋TSA with low （0.1 μmol/L＋0.05 μmol/ L）, middle （1 μmol/L＋0.5μmol/L） and high （10 μmol/L＋5 μmol/L） dose. Oral epithelial cells were taken as control. Methylation degree was detected by real-time PCR. The proliferation and apoptosis were detected by MTT and DAPI staining. The data were analyzed using SPSS 20.0 software package. RESULTS： Maspin methylation was enhanced in tumor cells when compared with control group （P〈0.01）. Maspin methylation was decreased in middle and high dose group when compared with negative group （P〈0.05, P〈0.01）. The inhibition ratio of cell multiplication was enhanced when compared with negative group （P〈0.05, P〈0.01）. The apoptosis content was enhanced after incubation with ADC＋TSA with a dose dependent manner （P〈0.05）. CONCLUSIONS： Methylation of Maspin gene may be involved in the proliferation of oral cancer cells.