叶酸对宫颈癌细胞生物学行为及自噬的影响  

Effect on biological behavior and autophagy of folic acid of cervical cancer cells

作　　者：刘雅淑 雷声云 郑春艳 陈茜[3] LIU Ya-shu;LEI Sheng-yun;ZHENG Chun-yan;CHEN Qian(Department of Gynecology,Xingyuan Hospital of Yulin,Yulin 719000,Shaanxi,China;Department of Gynecology and Obstetrics,Xi’an International Medical Center Hospital,Xi’an 719100,Shaanxi,China;The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,Shaanxi,China)

机构地区：[1]榆林市星元医院妇科,陕西榆林719000 [2]西安国际医学中心医院妇产科,陕西西安719100 [3]西安交通大学第二附属医院妇产科,陕西西安710004

出　　处：《生物医学工程与临床》2020年第4期375-380,共6页Biomedical Engineering and Clinical Medicine

摘　　要：目的探讨叶酸对宫颈癌细胞增殖、迁移和凋亡及细胞自噬的影响。方法以宫颈癌细胞株SiHa作为研究对象,用不同质量浓度梯度叶酸培养SiHa细胞,其中中等剂量10.0μg/mL为对照组,0.1μg/mL和1.0μg/mL为低叶酸组,100.0μg/mL和1000.0μg/mL为高叶酸组;用CCK-8法检测细胞的增殖活性,划痕实验检测细胞的迁移能力(划痕宽度占初始划痕宽度的比值:划痕宽度比值),流式细胞仪检测细胞的凋亡率;Western blot检测Beclin1、LC3和p62蛋白的表达水平。结果CCK-8法结果显示,质量浓度100.0μg/mL、1000.0μg/mL叶酸明显抑制SiHa细胞增殖活性(0.79±0.01、0.52±0.09 vs 1.24±0.03),且随着叶酸浓度增加,对宫颈癌SiHa细胞增殖的抑制作用越强。细胞划痕实验结果显示,随着叶酸质量浓度的增加,培养48 h的划痕宽度比值增加,叶酸质量浓度为1000.0μg/mL时划痕宽度比值比原始划痕宽度还宽且随着叶酸质量浓度的增加而增加(1.25±0.03 vs 0.59±0.06),对SiHa细胞迁移的抑制作用更强。细胞凋亡实验结果显示,100.0μg/mL、1000.0μg/mL质量浓度的叶酸显著提高SiHa细胞的凋亡率(12.74%±5.12%、26.62%±4.80%vs 6.80%±2.30%),且随着叶酸质量浓度增加,细胞凋亡率越大;随叶酸质量浓度的增加(≥1.0μg/mL),Beclin1、LC3蛋白表达水平升高,p62蛋白表达水平下降。结论叶酸质量浓度≥100.0μg/mL抑制宫颈癌SiHa细胞的增殖及迁移,促进其凋亡,且具有剂量依赖性,可能与细胞自噬的激活有关。Objective To investigate proliferation,migration,apoptosis and autophagy of folic acid on cervical cancer cells.Methods The cervical cancer cell line SiHa was cultured with different mass concentration of folic acid,with 10.0μg/mL as control group,0.1μg/mL and 1.0μg/mL as low folic acid group,100.0μg/mL and 1000.0μg/mL as high folic acid group.The CCK-8 method was used to detect cell proliferation activity,scratch experiment to detect cell migration ability(scratch width ratio of initial scratch width:scratch width ratio),and flow cytometry to detect cell apoptosis rate.The expression levels of Beclin1,LC3 and p62 protein were detected by Western blot.Results The results of CCK-8 method showed that concentration of 100.0μg/mL and 1000.0μg/mL folic acid significantly inhibited proliferation activity of SiHa cells(0.79±0.01,0.52±0.09 vs 1.24±0.03).The results of cell scratch experiment showed scratch width ratio at 48-hour in culture increased with concentration of folic acid.At 1000.0μg/mL folic acid concentration,the scratch width ratio was greater than original scratch width,and increased with the increasing of folic acid concentration(1.25±0.03 vs 0.59±0.06),and stronger inhibitory effect on SiHa cell migration was observed.The results of apoptosis experiments showed that 100.0μg/mL and 1000μg/mL folic acid concentration significantly increased apoptosis rate of SiHa cells(12.74%±5.12%,26.62%±4.80%vs 6.80%±2.30%),and apoptosis rate increased with the increasing of folic acid concentration.The expression levels of Beclin1,LC3 protein were increased,p62 protein was decreased with folic acid concentration increasing(≥1.0μg/mL).Conclusion It is demonstrated that folic acid concentration≥100.0μg/mL can inhibit proliferation and migration of cervical cancer SiHa cells,and promote apoptosis in dose-dependent manner,which may be related to autophagy activation.

关 键 词：叶酸 宫颈癌 细胞增殖 细胞迁移 细胞凋亡 自噬 

分 类 号：R737.33[医药卫生—肿瘤] R364[医药卫生—临床医学]