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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。
OBJECTIVE: To analyze the regulatory mechanism of Maspin gene methylation in the process of oral mucosal squamous cell carcinoma (SCC) proliferation and provide references for related research and clinical treatment. Methods: The squamous cell carcinoma HIOEC-B (a) P cell lines were randomly divided into 4 groups: low (0.1μmol / L + 0.05μmol / L), medium (1μmol / L + 0.5μmol / L) and high dose (10μmol / L + 5μmol / L). Normal oral mucosa epithelial cells were used as control group. Real-time quantitative PCR was used to detect the methylation level of Maspin gene in each group. The proliferation of each group was detected by MTT assay. Apoptosis was detected by 4 ’, 6-diamidino-2-phenylindole (DAPI) staining. Data were analyzed by one-way ANOVA using SPSS20.0 software package. Results: Compared with the control group, Maspin gene methylation was significantly increased (P <0.01). Compared with the negative control group, Maspin gene methylation was significantly attenuated in the medium and high dose groups (P <0.05, P <0.01). The inhibition rates of cell proliferation in low, medium and high dose groups were significantly higher than those in negative group (P <0.05, P <0.01). With the increase of drug dose, the degree of apoptosis gradually increased (P <0.05). Conclusion: Maspin gene methylation may be involved in the proliferation of oral squamous cell carcinoma cells.