Effects of lncRNA XIST-targeting microRNA 101/EZH2 on cell proliferation and migration of pancreatic cancer cells
10.3760/cma.j.cn115667-20191022-00094
- VernacularTitle:长链非编码RNA XIST靶向miR-101/EZH2对胰腺癌细胞增殖和迁移的影响
- Author:
Jie MIN
1
;
Lili CAO
;
Binbin SHEN
;
Yu ZHOU
;
Liang LI
;
Jun ZHOU
Author Information
1. 浙江省嘉兴市第一医院(嘉兴学院附属第一医院)肝胆外科，嘉兴 314000
- From:
Chinese Journal of Pancreatology
2020;20(3):200-206
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of long-chain non coding RNA (lncRNA) XIST on the proliferation and migration of pancreatic cancer PANC1 cells, and clarify the targeting relationship between lncRNA XIST and miR-101/enhancer of zeste homologz(EZH2).Methods:Ninety cases of pancreatic cancer surgically resected and pathologically confirmed in the first hospital of Jiaxing city from July 2010 to September 2018 and its corresponding paracarcinoma normal tissue were collected. PANC1 cells were divided into sh-XIST group, SH control group, MiR control group and miR-101 group. The expression of LncRNA XIST and miR-101 in pancreatic cancer tissue and PANC1 cells in each group were detected by fluorescence quantitative PCR. The relationship between the expression of LncRNA XIST and miR-101 and the clinicopathological parameters of tumor was analyzed. The proliferation and migration ability of PANC1 cells in each group were analyzed by CCK8 method and transwell chamber test. The EZH2 expression level of PANC1 cells in each group were analyzed by western blot. PANC1 cells in each group was inoculated into BALB/C nude mice with a cell density of 3×10 6 cells/100 μl and the tumor volume was measured. The relationship between LncRNA XIST and its miR-101 and targeting gene EZH2 were analyzed by bioinformatics and double luciferase reporter genes. Results:Compared with the paracancerous tissues, the level of LncRNA XIST in pancreatic cancer tissue was significantly increased 2.89±0.42 vs (1.12±0.22, P<0.05), and the level of miR-101 was significantly decreased 0.32±0.12 vs (1.25±0.22, P<0.05), and the differences were statistically significant ( P<0.05). LncRNA XIST expression in pancreatic cancer tissue was obviously increased along with higher differentiation degree, advanced TNM stage and lymph node metastasis, while miR-101 was greatly decreased. Compared with the cells in the sh-control group, the expression level of LncRNA XIST in the sh-XIST group was significantly decreased (0.34±0.18 vs 1.21±0.27). Compared with miR-control cells, the level of miR-101 cells in miR-101 group significantly increased (2.94±0.31 vs 1.54±0.29 ), and the differences were statistically significant ( P<0.05). After 72 h cell culture, the light absorption value at 450 nm in sh-control group, sh-XIST group, miR-control group and miR-101 group was 1.98±0.24, 1.21±0.20, 1.87±0.21 and 1.11±0.17; the number of transmembrane cells were (74.25±6.79 ), (29.11±5.17), (61.27±5.19) and (20.47±4.58)per 200 times visual field; the cell proliferation activity and migration ability in sh-XIST group were significantly decreased than sh-control group, miR-101 group and miR-control group and the xenograft tumor grew obviously slowly, all the differences were statistically significant (all P<0.05). Conclusions:LncRNA XIST can target miR-101/EZH2, regulating the proliferation and migration of pancreatic cancer cells, which promotes the occurrence and development of pancreatic cancer.
