miR-182通过HES1调控宫颈癌耐放疗细胞株的上皮间质转化

miR-182 attenuates EMT of radioresistant cervical cancer cells by targeting HES1

目的探讨miR-182在宫颈癌耐放疗细胞株中的作用及其调控方式。方法采用分割剂量照射法诱导宫颈癌耐放疗HeLa细胞株和SiHa细胞株,分别每周4 Gy照射1次(HR4组和SR4组)和每周6 Gy照射1次(HR6组和SR6组)。采用RT-qPCR和Western blot检测4株耐放疗细胞中发状分裂相关增强子1(hairy and enhancer of split 1,HES1)和miR-182的表达。在筛选的耐放疗HR6细胞中下调miR-182后,检测该细胞中上皮间质转化相关因子E-cadherin、Slug mRNA和蛋白的表达,Transwell小室法检测细胞迁移和侵袭能力;并观察耐放疗HR6细胞中miR-182与HES1相互表达的影响。结果与未照射宫颈癌HeLa和SiHa细胞比较,耐放疗HeLa和SiHa细胞中HES1表达均显著降低(P<0.05),miR-182表达显著升高(P<0.05),且在不同耐放疗细胞株中miR-182表达有差异性(P<0.05)。下调miR-182后,HR6细胞中E-cadherin表达增强,Slug表达降低,且细胞迁移、侵袭能力减弱。HR6细胞中miR-182下调后HES1表达升高,但下调HES1后miR-182表达无明显变化。结论宫颈癌耐放疗细胞中下调miR-182可上调HES1,从而逆转上皮间质转化并抑制细胞迁移、侵袭,HES1可能是miR-182发挥作用的靶点。

Objective To evaluate the effect of miR-182 in radioresistant cervical caner cells and its probable regulative mode.Methods HeLa cell lines and SiHa cell lines were induced by fractionated dose irradiation,respectively,once a week at 4 Gy（HR4 group and SR4 group） and once a week at 6 Gy（HR6 group and SR6 group）. Expression of HES1 and miR-182 in 4 radiation resistant cells were detected using RT-qPCR and Western blotting.The expression levels of epithelial-mesenchymal transition-（EMT-） related factors（E-cadherin,Slug） were detected after down-regulation of miR-182 in selected radioresistant HR6 cells,and the capability of invasion and migration of HR6 cells was measured using the transwell assay. Additionally,we separately knocked down miR-182 or HES1 and investigated their ralationship. Results HES1 were significantly reduced in HeLa and SiHa cells after radiation,while miR-182 were significantly increased. Levels of miR-182 were diverse in different radioresistant cells. Knockdown of miR-182 significantly increased E-cadherin,decreased Slug in HR6 HeLa cells,and silenced their migration and invasion. The expression of HES1 in HR6 cells was increased after miR-182 knockdown,while the knockdown of HES1 did not affect miR-182. Conclusions Silencing miR-182 can effectively inverse EMT and inhibit migration and invasion by radioresistant cervical cancer cells. miR-182 may exert these effects by regulating the expression of HES1.