LncRNA ANRIL target miR-195 experimental study of radiation sensitivity of HCT116 cells and nude mouse transplant tumors
10.3760/cma.j.issn.1004-4221.2019.11.013
- VernacularTitle: LncRNA ANRIL靶向miR-195对HCT116细胞及裸鼠移植瘤放射增敏实验研究
- Author:
Xiaoyan CHEN
1
;
Chenbin WU
2
;
Xin TIAN
3
;
Xiaoli GOU
3
;
Yingqiang WU
4
;
Kui ZHAO
1
;
Rui XIE
1
Author Information
1. Department of Gastroenterology, Affiliated Hospital of Zunyi Medical University, Zunyi 563000, China
2. Tumor Radiotherapy Department, 95th Hospital of Liberation Army, Putian 351100, China
3. Department of Oncology, Hospital of Zunyi Medical University, Zunyi 563000, China
4. Department of General Medicine, Hospital of Zunyi Medical University, Zunyi 563000, China
- Publication Type:Journal Article
- Keywords:
ANRIL gene;
miR-195 gene;
HCT116 cell line;
Nude mouse transplant tumors;
Radiosensitivity
- From:
Chinese Journal of Radiation Oncology
2019;28(11):858-861
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and mechanism of LncRNA ANRIL on the radiosensitivity of HCT116 cells line and nude mouse transplant tumors.
Methods:The expression of LncRNA ANRIL in colorectal cancer cells was detected by qPCR. The negative control siRNA, ANRIL siRNA, miR-NC mimic, miR-195 mimic, miR-NC inhibitor and miR-195 inhibitor were transfected into HCT116 cells, and marked as negative control group, silencing ANRIL group, overexpressing miR-NC group, overexpressing miR-195 group, inhibiting miR-NC group and inhibiting miR-195 group, and the HCT116 cells without any treatment were marked as the blank control group. The clone formation assay was used to detect radiosensitivity of colorectal cancer cells, flow cytometry was used to detect apoptosis. The web site, StarBase, was used to predict the downstream miRNAs of ANRIL and dual luciferase reporter gene assay was used to further verify. Subcutaneous tumor transplantation assay was used to detect the effect of ANRIL on the growth of colorectal cancer cells after irradiation.
Results:After irradiation with 2, 4, 6 and 8 Gy, the cell survival fraction of silencing ANRIL group was significantly decreased when compared with that of negative control group (P<0.05), and the radiosensitivity ratio was 1.52. The apoptosis rate of the silencing ANRIL+ 4 Gy group was significantly higher than that of the negative control+ 4 Gy group ((27.86±2.78)% vs. (12.06±1.46)%, P<0.05). The results of the experiment on nude mouse transplant tumors showed that the tumor volume in the negative control group was lower than that of the silent ANRIL group on days 13, 16, 19, 22 and 25 ((234±66) mm3, (273±63) mm3, (296±72) mm3, (321±85) mm3 and (403±94) mm3 vs. (357±79) mm3, (485±124) mm3, (617±143) mm3, (764±174) mm3 and (985±221) mm3P<0.05). MiR-195 is a target gene of ANRIL, and inhibition of miR-195 can reverse the inhibitory effect of silencing ANRIL on radiosensitivity, apoptosis and xenografts of HCT116 cells.
Conclusions:LncRNA ANRIL regulates the radiosensitivity of colorectal cancer cells by miR-195, which may provide a new sensitizing target for clinical colorectal cancer radiotherapy.
