EPB41L3基因甲基化参与调控宫颈癌进展的相关性研究<sup>★</sup>

doi:10.3969/j.issn.2095-1264.2024.05.06

首页 > 过刊浏览>2024年第5期 >554-559. DOI:10.3969/j.issn.2095-1264.2024.05.06

EPB41L3基因甲基化参与调控宫颈癌进展的相关性研究★
DOI:
                        10.3969/j.issn.2095-1264.2024.05.06
                    
作者:
                        
                        
                    
作者单位:1.新疆医科大学附属肿瘤医院，新疆 乌鲁木齐，830000;2.新疆医科大学第一附属医院，新疆 乌鲁木齐，830054
作者简介:张媛媛，女，博士，主任医师，研究方向为妇科肿瘤。
通讯作者:古扎丽努尔·阿不力孜，女，博士，主任医师，研究方向为妇科肿瘤。
中图分类号:R737.33
基金项目:★新疆自治区自然科学基金项目（2021D01C404）；“天山英才”医药卫生高层次人才培养项目（TSYC202301B076）。

Correlation between methylation of EPB41L3 gene and regulation of cervical cancer progression^★

Author:

Affiliation:

1.Cancer Hospital Affiliated to Xinjiang Medical University, Urumqi, 830000, Xinjiang, China;2.The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, Xinjiang, China

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摘要:

目的探究EPB41L3基因甲基化与宫颈癌进展的相关性。方法收集临床慢性宫颈炎、高级别宫颈上皮内瘤变及宫颈癌患者的宫颈组织，用基因甲基化特异性PCR扩增（MSP）试剂盒检测样本中EPB41L3基因的甲基化水平；另外选取宫颈癌HeLa细胞和SiHa细胞，以人正常宫颈上皮细胞HUCEC作为对照，PCR检测细胞中EPB41L3基因甲基化水平，RT-PCR和免疫印迹检测EPB41L3基因表达水平；根据是否使用去甲基化药物5-氮杂-2'-脱氧胞苷（5AZA）将细胞分为4组：HeLa+Control组、HeLa+5AZA组、SiHa+Control组、SiHa+5AZA组。采用CCK8实验检测各组细胞的增殖率，Transwell实验检测各组细胞的迁移率，流式细胞术检测各组细胞的凋亡率，进行组间对比分析。结果慢性宫颈炎、高级别宫颈上皮内瘤变、宫颈癌组织中EPB41L3甲基化百分比分别为6.67% 、73.33%、93.33%。在HeLa细胞和SiHa细胞中均检测到EPB41L3甲基化。EPB41L3基因在HeLa细胞和SiHa细胞中的表达水平均显著低于对照组（P<0.000 1， P=0.002 5）。HeLa+5AZA组细胞增殖率显著低于HeLa+Control组（P=0.003 5），SiHa+5AZA组细胞增殖率显著低于SiHa+Control组（P=0.006 0）。HeLa+5AZA组细胞迁移率显著低于HeLa+Control组（P=0.000 9），SiHa+5AZA组细胞迁移率显著低于SiHa+Control组（P=0.001 9）。HeLa+5AZA组细胞凋亡率显著高于HeLa+Control组（P=0.002 4），SiHa+5AZA组细胞凋亡率显著高于SiHa+Control组（P=0.002 4）。结论 EPB41L3基因甲基化后可能失去对宫颈癌细胞的抑制作用，其表达水平与宫颈癌进展可能呈正相关。

Abstract:

Objective To investigate the correlation between EPB41L3 gene methylation and cervical cancer progression.Methods Cervical tissues of patients with clinical chronic cervicitis, cervical intraepithelial neoplasia and cervical cancer were collected. The methylation of EPB41L3 gene in the samples was detected by gene methylation-specific PCR amplification (MSP) kit. Taking the normal cervical epithelial cells as control, the methylation of EPB41L3 gene in HeLa and SiHa cervical cancer cell lines were detected by PCR. The expression levels of EPB41L3 gene in cervical cancer cell lines and normal cervical epithelial cell lines were detected by RT-PCR and western blotting. Cells were divided into four groups according to whether the demethylated drug 5-aza-2'-deoxygcitidine (5AZA) was used: HeLa group, HeLa+5AZA group, SiHa group, and SiHa+5AZA group. In addition, CCK8 assay was used to detect cell proliferation rate, Transwell assay was used to detect cell mobility, flow cytometry was used to detect cell apoptosis rate in each group. Comparative analysis was performed between the groups.Results The methylation percentage of EPB41L3 in chronic cervicitis, cervical intraepithelial neoplasia and cervical cancer was 6.67%, 73.33% and 93.33%, respectively. EPB41L3 methylation was detected in HeLa and SiHa cells. The expression of EPB41L3 in HeLa and SiHa cells was significantly lower than that in the control group (P<0.000 1, P=0.002 5). The cell proliferation rate of HeLa+5AZA group was significantly lower than that of HeLa+Control group (P=0.003 5), and that of SiHa+5AZA group was significantly lower than that of SiHa+Control group (P=0.006 0). The cell migration rate of HeLa+5AZA group was significantly lower than that of HeLa+Control group (P=0.000 9), and that of SiHa+5AZA group was significantly lower than that of SiHa+Control group (P=0.001 9). The cell apoptosis rate of HeLa+5AZA group was significantly higher than that of HeLa+Control group (P=0.002 4), and that of SiHa+5AZA group was significantly higher than that of SiHa+Control group (P=0.002 4).Conclusion EPB41L3 gene may lose its inhibitory effect on cervical cancer cells after methylation, and its expression level may be positively correlated with the progression of cervical cancer.