Influence of miR-21 on radiation sensitivity of cervical cancer HeLa cells
10.13481/j.1671-587x.20140603
- VernacularTitle:miR-21对宫颈癌HeLa细胞辐射敏感性的影响
- Author:
Shan XU
;
Mengmeng YAN
;
Yang LIU
;
Pengwu LI
;
Shumei MA
;
Xiaodong LIU
- Publication Type:Journal Article
- Keywords:
miR-21;
cervical neoplasms;
HeLa cells;
radiation sensitivity;
apoptosis;
autophagy
- From:
Journal of Jilin University(Medicine Edition)
2014;(6):1138-1143
- CountryChina
- Language:Chinese
-
Abstract:
Objective To research the relationship between miR-21 and radiation sensitivity of cervical cancer HeLa cells,and to elucidate the possible action mechanism of miR-21 in apoptosis and autophagy of cells.Methods The HeLa cells were divided into mock groups (transfected with miR-21 NC,mimics,inhibitor NC or inhibitor)and 4 Gy irradiation groups (transfected with miR-2 1 NC, mimics,inhibitor NC or inhibitor with irradiation). The miR-2 1 expression was detected by real-time PCR;colony-forming assay was used to detect the changes of radiation sensitivity;the apoptosis and autophagy were analyzed by FACS assay;the target gene of miR-2 1 was predicted by bioinformatics methods;Dual-luciferase Reporter Assay was used to demonstrate the binding of miR-2 1 and targets;the expression of beclin1 protein was measured by Western blotting method. Results After 4 Gy radiation,compared with mock group,the expression of miR-21 was increased remarkably (P<0.05). Colony-forming assay showed that there were no changes of radiation sensitivity of HeLa cells after overexpressing or inhibiting miR-21 compared with miR-21 NC or miR-21 inhibitor NC groups(P>0.05).Compared with miR-21 NC+4 Gy group,the apoptotic rate of HeLa cells in miR-21 mimics+4 Gy group was increased (P<0.05).After transfection with miR-21 inhibitor,compared with miR-21 inhibitor NC group,the apoptotic rate of HeLa cells in duced by radiation was decreased (P<0.05 ). Compared with miR-21 NC group, the autophagy rate was increased after transfection with miR-21 mimics (P<0.05).After transfection with miR-21 mimics and radiation, the expression level of beclin1 protein was increased (P<0.05 ) compared with miR-21 NC+4 Gy group. Conclusion Beclin1 is the target gene of miR-21 in HeLa cells. Overexpression of miR-21 can increase the radiation-induced apoptosis and autophagy,but has on influence in radiation sensitivity of HeLa cells.
