卷曲乳酸杆菌对宫颈癌细胞迁移、侵袭的影响及其机制研究

Effect of Lactobacillus crimpus on migration and invasion of cervical cancer cells and its mechanism

目的:探讨卷曲乳酸杆菌对宫颈癌细胞迁移、侵袭能力的影响及其机制。方法:将卷曲乳酸杆菌与宫颈鳞癌SiHa细胞在合适条件下共培养,Transwell实验观察共培养前、后细胞迁移和侵袭能力的变化。分别采用实时荧光定量PCR(RT-qPCR)、western blotting法检测ABCG2、PCNA、ATM、TDG、LIG1、OGG1、HMGB1基因和蛋白表达。结果:共培养18 h和24 h后,卷曲乳酸杆菌能有效抑制SiHa细胞的迁移和侵袭(P<0.001)。同时,与宫颈癌相关的致癌基因PCNA、ATM和LIG1表达水平在共培养12 h后开始显著下降(P<0.05);OGG1和HMGB1表达水平在共培养18 h后开始出现显著下降(P<0.05);ABCG2和TDG表达水平在共培养前、后未出现明显变化(P>0.05)。另外PCNA、OGG1和HMGB1蛋白表达水平在共培养12 h后开始呈现显著性下降(P<0.05),ABCG2、LIG1和ATM蛋白表达水平在共培养18 h后开始出现显著下降(P<0.05),TDG蛋白表达水平在共培养前、后未出现明显变化(P>0.05)。结论:卷曲乳酸杆菌可以有效抑制宫颈癌SiHa细胞的迁移和侵袭,并抑制PCNA、ATM、LIG1、OGG1、HMGB1等致癌基因的表达。

Objective:To study the effect of Lactobacillus crimpus on the migration and invasion ability of cervical cancer cells and its mechanism.Methods:Lactobacillus crimpus was co-cultured with SiHa cells of cervical squamous cell carcinoma under suitable conditions.Transwell test was used to observe the changes of cell migration and invasion ability before and after co-culture.Meanwhile,real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and western blotting were used to detect the gene and protein expressions of ABCG2,PCNA,ATM,TDG,LIG1,OGG1 and HMGB1.Results:Lactobacillus crimpus could effectively inhibit the migration and invasion of SiHa cells after co-culture for 18 h and 24 h(P<0.001).At the same time,the oncogenes PCNA,ATM and LIG1 associated with cervical cancer began to decrease significantly after 12 h co-culture(P<0.05);OGG1 and HMGB1 decreased significantly after co-culture for 18 h(P<0.05);no significant changes were observed between ABCG2 and TDG before and after co-culture(P>0.05).In addition,the expressions of PCNA,OGG1 and HMGB1 proteins began to decrease significantly after 12 h co-culture(P<0.05),ABCG2,LIG1 and ATM proteins began to decrease significantly after 18 h co-culture(P<0.05),and there were no significant changes in the relative protein expression levels before and after TDG co-culture(P>0.05).Conclusion:Lactobacillus crimpus can effectively inhibit the migration and invasion of SiHa cells of cervical cancer and inhibit the expressions of PCNA,ATM,LIG1,OGG1,HMGB1 and other oncogenes.