miR-204在宫颈癌中的表达及对宫颈癌细胞生物学行为的调控机制研究

The expression of miR-204 in cervical cancer and its regulatory mechanism on the biological behavior of cervical cancer cells

目的观察miR-204在宫颈癌中的表达情况并探讨调控宫颈癌细胞生物学行为的影响机制。方法RTPCR分别检测30例宫颈癌组织和癌旁组织miR-204mRNA表达情况,将Hela细胞株细胞分为对照组、miR-204过表达组、miR-204沉默组、miR-204过表达+SIRT1激动剂白藜芦醇组、miR-204沉默+SIRT1拮抗剂EX527组;用MTT法检测细胞活力,Annexin V-FIPC/PI双染色法流式细胞仪检测细胞凋亡,Transwell小室细胞迁移实验及体外侵袭实验。RT-PCR及Western blot检测细胞SIRT1、CXCR4mRNA和蛋白表达情况。结果与癌旁正常组织相比,宫颈癌组织中miR-204mRNA相对表达量明显下降,SIRT1和CXCR4mRNA相对表达量明显增加,差异有统计学意义(P<0.05)。与对照组相比,miR-204过表达组细胞增殖率、细胞侵袭数、SIRT1和CXCR4显著下降,凋亡率显著上升(P<0.05),miR-204过表达+白藜芦醇组增殖率无明显差异(P>0.05),miR-204过表达+EX527组变化更为显著,且miR-204过表达+EX527组与miR-204过表达组存在显著性差异(P<0.05)。结论miR-204在宫颈癌中呈现低表达,提高miR-204表达可以经SIRT1/CXCR4通路抑制宫颈癌细胞增殖和侵袭,并促进凋亡。

Objective To observe the expression of miR-204 in cervical cancer and explore the regulating mechanism of cervical cancer cells.Methods RT-PCR was used to detect the expression of miR-204 mRNA in 30 cases of cervical cancer tissues and adjacent tissues,Hela cell line were divide into control group,miR-204 overexpression group,miR-204 silence group,miR-204 overexpression+SIRT1 agonist resveratrol group,miR-204 silence group+SIRT1 antagonist EX527 group;MTT,Annexin V-FIPC/PI double staining,Transwell chamberwere used to detect cell viability,apoptosis by flow cytometry and invasion ability.RT-PCR and Western blot were used to detect the expression of SIRT1 and CXCR4 mRNA and protein.Results Compared with normal tissues adjacent to cancer,the relative expression of miR-204 mRNA in cervical cancer tissue was significantly decreased,and the relative expression of SIRT1 and CXCR4 mRNA was significantly increased(P<0.05).Compared with the control group,the cell proliferation rate,cell invasion number,SIRT1 and CXCR4 of the miR-204 overexpression group decreased significantly,and the apoptosis rate increased significantly(P<0.05).There was no significant difference in the proliferation rate of miR-204 overexpression+resveratrol group(P>0.05).The changes in the miR-204 overexpression+EX527 group were more significant,and there was a significant difference between the miR-204 overexpression+EX527 group and the miR-204 overexpression group(P<0.05).Conclusion The expression of miR-204 is low in cervical cancer,increasing the expression of miR-204 can inhibit the proliferation and invasion of cervical cancer cells through the SIRT1/CXCR4 pathway,and promote apoptosis.