Effects and related mechanisms of long non-coding RNA AC092295.2 on proliferation and invasion abilities of endometrial cancer cells
10.3760/cma.j.cn115355-20240315-00117
- VernacularTitle:长链非编码RNA AC092295.2对子宫内膜癌细胞增殖和侵袭能力的影响及相关机制
- Author:
Xiaojie XUE
1
;
Feirong LI
;
Haiying DAI
;
Duan CHEN
;
Hui LIU
;
Quan LI
Author Information
1. 黄石市中心医院（湖北理工学院附属医院）检验科 肿瘤分子诊断与治疗黄石市重点实验室，黄石 435000
- Publication Type:Journal Article
- Keywords:
Endometrial neoplasms;
Long non-coding RNA;
MicroRNAs;
Cell proliferation;
Neoplasm invasiveness
- From:
Cancer Research and Clinic
2025;37(2):81-86
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects and related mechanisms of long non-coding RNA (lncRNA) AC092295.2 expression on the proliferation and invasion abilities of endometrial cancer cells.Methods:The correlations between AC092295.2 expression level and overall survival (OS) and disease-free survival (DFS) of endometrial cancer patients (180 cases) were analyzed by cBioPortal database (updated in 2022). The miRNA with complementary nucleotide sequences to AC092295.2 was predicted by DIANA Tools sequencing tool. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression of AC092295.2 in human immortalised endometrial epithelial cell line hEEC and human endometrial cancer cell lines (KLE, AN3CA, Ishikawa, HEC-1A). Ishikawa cells with the lowest expression level of AC092295.2 were selected and divided into AC092295.2 overexpression group and control group, which were transfected with pGEX-AC092295.2 plasmid and pGEX-NC plasmid, respectively. qRT-PCR was used to detect the relative expression levels of AC092295.2 and miRNA-200c-3p (miR-200c-3p) in the two groups of cells. Methyl thiazol tetrazolium (MTT) method was used to detect cell viability. Transwell assay was used to detect cell invasion ability. Western blotting was used to detect the expression of proteins related to PTEN-AKT pathway. The dual luciferase reporter gene assay was used to verify the targeting relationship between AC092295.2 and miR-200c-3p.Results:The analysis results of cBioPortal database showed that the OS and DFS of endometrial cancer patients with high expression of AC092295.2 were better than those of patients with low expression (both P < 0.001); the expression levels of AC092295.2 and miR-200c-3p in endometrial cancer tissues were negatively correlated ( r = -0.846, P < 0.001). The results of qRT-PCR detection showed that the relative expression levels of AC092295.2 in endometrial cancer cell lines KLE, AN3CA, Ishikawa, HEC-1A and immortalized endometrial epithelial cell line hEEC were 0.56±0.09, 0.68±0.06, 0.17±0.07, 0.49±0.12 and 0.99±0.11, respectively, and the difference was statistically significant ( F = 35.10, P < 0.001); the relative expression levels of AC092295.2 in Ishikawa cells in the AC092295.2 overexpression group and the control group were 8.92±1.78 and 1.06±0.39, respectively, and the difference was statistically significant ( t = 4.31, P = 0.005). MTT assay results showed that the cell viability of Ishikawa cells in the AC092295.2 overexpression group was lower than that in the control group on days 2, 3, 4, and 5 (all P < 0.05). Transwell assay results showed that the number of invasive Ishikawa cells in the AC092295.2 overexpression group and the control group were 73±4 and 135±14, respectively, and the difference was statistically significant ( t = 4.25, P = 0.005). Western blotting results showed that the relative expression level of phosphatase and tensin homolog (PTEN) protein in Ishikawa cells in the AC092295.2 overexpression group was higher than that in the control group, while the relative expression levels of phosphorylated protein kinase B (p-AKT), Yes associated protein (YAP), murine double mimute 2 (MDM2), and bcl-2-associated death-promoting factor (BAD) proteins were lower than those in the control group. Dual luciferase reporter gene assay and qRT-PCR verified that AC092295.2 targeted negative regulation of miR-200c-3p expression. Conclusions:AC092295.2 is lowly expressed in endometrial cancer cells. Overexpression of AC092295.2 can inhibit the proliferation and invasion abilities of endometrial cancer cells, and its mechanism may be related to the expression of miR-200c-3p-PTEN-AKT signaling pathway.
