Effects of long-chain non-coding RNA U73166 on proliferation and invasion of lung cancer cells by targeting miR-618 and its mechanism
10.3760/cma.j.cn121382-20250107-00005
- VernacularTitle:长链非编码RNA U73166通过调控 miR-618对肺癌细胞增殖和侵袭的影响及其机制
- Author:
Zhenzhen LIU
1
;
Wei LIU
;
Lina GUAN
;
Nan WU
Author Information
1. 山东大学附属威海市立医院呼吸与危重症医学科，威海 264200
- Keywords:
Lung cancer;
Long-chain non-coding RNA;
U73166;
microRNA-618;
Janus kinase 2;
Signal transducer and activator of transcription 3;
Signal-transducing adaptor
- From:
International Journal of Biomedical Engineering
2025;48(3):264-270
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze the expression of long-chain non-coding RNA U73166 in lung cancer tissues and its relationship with patients′ prognosis, and to explore the effects of silencing U73166 on proliferation and invasion of lung cancer H1299 cells and its regulatory mechanism. Methods:The expression level of U73166 in lung cancer tissues and normal tissues, as well as its correlation with lung cancer patients′ overall survival, were analyzed using the gene expression profiling interactive analysis (GEPIA) database. After culturing, H1299 cells were divided into a control group and a transfection group based on treatment conditions, and were transfected with 25 μmol/L of U73166 negative control and U73166 inhibitor, respectively. The effects of silencing U73166 on the relative expression of U73166 and microRNA-618 ( miR-618) genes in H1299 cells were assessed by real-time reverse transcription-PCR method. A cell counting kit-8 assay was used to evaluate the impact of silencing U73166 on the viability of H1299 cells. A transwell invasion assay was performed to detect the invasive ability of H1299 cells. The Linc2GO database and a dual-luciferase reporter assay were used to predict and verify the binding site between U73166 and miR-618. Western blotting was used to analyze the relative expression of phosphorylated Janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), and phosphorylated signal-transducing adaptor molecule 1 (p-STAM1) in the JAK2/STAT3 signaling pathway to evaluate the effects of silencing U73166 on this pathway in H1299 cells. Data were analyzed by an independent sample t test or one-way analysis of variance. Results:Analysis of the GEPIA database revealed that U73166 relative expression level in lung cancer tissues ( n=383) was significantly higher than that in normal tissues ( n=347) ( P<0.01). The overall survival of lung cancer patients with low U73166 expression [(245±2) months] was longer than that of patients with high U73166 expression [(167±2) months] ( P<0.05). The relative expression of U73166 were 7.81±0.99 in the control group and 1.01±0.26 in the transfection group, respectively, and the relative expression of miR-618 were 1.03±0.20 in the control group and 4.83±1.27 in the transfection group, respectively. Silencing U73166 significantly downregulated its expression ( t=6.66, P<0.01) and upregulated the relative expression of miR-618 ( t=2.96, P<0.01) in H1299 cells. After silencing U73166, the absorbance values of H1299 cells in the transfection group on days 2, 3, 4, and 5 (0.36±0.04, 0.74±0.05, 1.07±0.09, and 1.18±0.10) were significantly lower than those in the control group (0.55±0.03, 1.20±0.08, 1.63±0.07, and 1.90±0.07) ( P<0.05, 0.01). The number of invasive cells in the control and transfection groups were 52.03±6.08 and 19.92±3.78, respectively. There were significantly fewer invasive cells in the transfection group ( t=4.49, P<0.01). After transfection with wild-type U73166, the relative luciferase activity in the miR-618 group (0.32±0.05) was significantly lower than that in the miR-negtive control group (0.96±0.15) ( t=4.02, P<0.01). However, after transfection with mutant U73166, there was no statistically significant difference in relative luciferase activity between the miR-618 group (1.01±0.15) and the miR-negtive control group (1.03±0.11) ( t=0.09, P>0.05). The relative expression of p-JAK2, p-STAT3, and p-STAM1 proteins in the transfection group were 2.08±0.21, 1.36±0.20, and 0.55±0.12, respectively. These values were significantly lower than those in the control group (3.72?±?0.29, 5.56?±?0.19, and 4.38±0.17) (all P<0.01). Conclusions:U73166 is highly expressed in lung cancer tissues and lung cancer cells, and its expression is related to lung cancer patients′ overall survival. Silencing U73166 can target miR-618, which inhibits the proliferation and invasion of H1299 cells.
