LncRNA LINC00525调控miR-338-3p/RUNX2轴对子宫颈癌细胞生物学行为的影响

Effects of LncRNA LINC00525 on the Biological Behavior of Cervical Cancer Cells by Regulating MiR-338-3p/RUNX2 Axis

目的:探讨长链非编码RNA长基因非蛋白质编码RNA 525(LncRNA LINC00525)调控miR-338-3p/核心结合因子(RUNX2)轴对子宫颈癌细胞生物学行为的影响。方法:样本选自2018年10月至2020年10月期间石家庄市第四医院的52例子宫颈癌组织和52例癌旁组织,以及子宫颈癌细胞HeLa、C33A、SiHa和人正常子宫颈上皮细胞Ect1/E6E7。采用实时荧光定量PCR(qRT-PCR)检测LINC00525、miR-338-3p、RUNX2 mRNA的表达;通过细胞计数试剂8(CCK8)、Transwell、蛋白印迹(Western blot)实验检测HeLa细胞的增殖、迁移与侵袭、RUNX2蛋白的表达;采用双荧光素酶报告基因、RNA下拉实验验证LINC00525与miR-338-3p、miR-338-3p与RUNX2的关系。结果:①与癌旁组织比较,子宫颈癌组织中LINC00525、RUNX2 mRNA表达水平显著升高(P<0.05),miR-338-3p表达水平显著降低(P<0.05);与人正常子宫颈上皮细胞Ect1/E6E7比较,子宫颈癌HeLa、C33A、SiHa细胞中LINC00525、RUNX2 mRNA表达水平显著升高(P<0.05),miR-338-3p表达水平显著降低(P<0.05),且HeLa细胞变化最显著。②下调LINC00525抑制HeLa细胞增殖、迁移与侵袭。③LINC00525靶向负调控miR-338-3p的表达;miR-338-3p靶向下调RUNX2的表达;④下调miR-338-3p表达减弱了下调LINC00525对HeLa细胞增殖、迁移与侵袭的抑制作用,过表达RUNX2减弱了下调LINC00525或过表达miR-338-3p对HeLa细胞增殖、迁移与侵袭的抑制作用。结论:下调LINC00525可能通过调控miR-338-3p/RUNX2轴抑制子宫颈癌细胞增殖、迁移与侵袭。

Objective:To investigate the effect of long non-coding RNA,long intergenic non-protein coding RNA 525(LncRNA LINC00525)on the biological behavior of cervical cancer cells by regulating miR-338-3p/core binding factor-α(RUNX2)axis.Methods:We included samples from 52 cases of cervical cancer tissue and 52 cases of paracarcinoma tissue from October 2018 to October 2020 in Shijiazhuang Fourth Hospital,and cell lines including cervical cancer cells HeLa,C33A,SiHa and human normal cervical epithelial cells Ect1/E6E7.Expressions of LINC00525,miR-338-3p and RUNX2 mRNA were detected by qRT-PCR.The proliferation,migration,invasion and the RUNX2 protein expression of HeLa cells were detected by CCK8,Transwell and Western blot,respectively.Dual luciferase reporter assay and RNA pull down were used to verify the relationship between miR-338-3p and LINC00525,RUNX2.Results:①Compared with paracancer tissues,cervical cancer tissues showed significantly higher expression levels of LINC00525 and RUNX2 mRNA(P<0.05)and lower expression level of miR-338-3p(P<0.05).Compared with Ect1/E6E7 in human normal cervical epithelial cells,expression levels of LINC00525 and RUNX2 mRNA in HeLa,C33A and SiHa cells were significantly increased(P<0.05),while the expression level of miR-338-3p was significantly decreased(P<0.05).HeLa cells showed the most significant changes.②Silencing LINC00525 inhibited the proliferation,migration and invasion of HeLa cell.③LINC00525 negatively regulates the expression of miR-338-3p which down regulates the expression of RUNX2.④Down-regulating of miR-338-3p expression weakened the inhibitory effect of silenced LINC00525 on the proliferation,migration and invasion of HeLa cells,while overexpression of RUNX2 attenuates the inhibitory effects of silenced LINC00525 or overexpressed miR-338-3p on the proliferation,migration,and invasion of HeLa cells.Conclusions:Silencing LINC00525 may inhibit the proliferation,migration and invasion of cervical cancer cells by regulating the miR-338-3p/RUNX2 axis.