POLQ对SACC-83细胞的影响及POLQ基因表达调控因子预测

The effects of POLQ on SACC-83 cells and prediction of POLQ gene regulator

目的:研究POLQ在肿瘤中的表达及对唾液腺腺样囊性癌细胞SACC-83增殖及克隆形成能力的影响,预测POLQ的表达调控因子。方法:应用数据库分析POLQ在肿瘤中的表达,并预测与POLQ启动子结合的FOXM1及二者的表达相关性。将体外培养的SACC-83细胞分为2组,用基因沉默技术(siRNA)抑制实验组细胞中POLQ的表达,未处理的细胞作为对照组。CCK-8实验检测增殖能力,平板克隆实验检测克隆形成能力,应用qRT-PCR检测基因表达水平。结果:数据库分析显示,多种肿瘤中POLQ表达高于正常组织。实验组细胞增殖能力及克隆形成能力均低于对照组(P<0.001,P<0.05)。FOXM1与POLQ启动子有5个结合位点,并与POLQ表达正相关(P<0.05)。PTEN下调促进SACC-83 POLQ与FOXM1表达(P<0.001,P<0.05)。结论:POLQ促进肿瘤的发生发展;促进SACC-83增殖及克隆形成能力;FOXM1可能正调控POLQ表达;PTEN负调控SACC-83 POLQ与FOXM1表达。

Objective:To investigate the expression of POLQ in tumors and its effects on cell proliferation and clone formation of salivary adenoid cystic carcinoma SACC-83 cells,and to predict and the transcription factor FOXM1 of POLQ.Methods:The expression of POLQ in tumors and the binding sites of FOXM1 in POLQ promoter were obtained by data base.In vitro cultured SACC-83 cells were treated by siRNA method for down-expression of POLQ as the cells of test group and those without treatment as the control group.CCK-8 assay was used to detect the proliferation of SACC cells,and clone formation assay was carried out to detect colongenesis of the cells.qRT-PCR was used to detect gene expression levels.Results:Data base showed that POLQ was up-expressed in many tumors(P<0.01).The proliferation and clonogenesis of the test group were lower than those of the control group of SACC-83 cells(P<0.001 and P<0.05).5 binding sites of FOXM1 in POLQ promoter sequence were predicted,and FOXM1 expression was positively correlated with POLQ in tumors(P<0.05).PTEN low-expression promoted POLQ and FOXM1 expression in SACC-83 cells(P<0.001,P<0.05).Conclusion:POLQ may promote tumor progeression,increase the proliferation and clone formation of SACC cells.FOXM1 is positively expressed with POLQ in tumors;PTEN is negatively regulated with POLQ and FOXM1 expression in SACC-83 cells.