Molecular mechanism of long non-coding RNA titin antisense RNA 1 regulating microRNA-134-5p/epidermal growth factor receptor axis in breast cancer cell glycolysis
- VernacularTitle:长链非编码RNA肌联蛋白反义RNA1调控微小RNA-134-5p/表皮生长因子受体轴对乳腺癌细胞糖酵解的分子机制
- Author:
Gaojin ZHOU
1
;
Liqing SHANG
;
Jun YAN
Author Information
- Keywords: breast cancer; long non-coding RNA; titin antisense RNA 1; microRNA-134-5p; epidermal growth factor receptor; glycolysis; cell proliferation; cell apoptosis
- From: Journal of Clinical Medicine in Practice 2025;29(6):62-68
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the molecular mechanism by which long non-coding RNA(lncRNA)titin antisense RNA 1(TTN-AS1)regulates the microRNA(miR)-134-5p/epidermal growth factor receptor(EGFR)axis in breast cancer cell proliferation,apoptosis and glycolysis.Methods Western blot was used to detect EGFR protein expression.Quantitative real-time poly-merase chain reaction(qRT-PCR)was employed to measure the expression levels of lncRNA TTN-AS1,miR-134-5p and EGFR mRNA in breast epithelial cells MCF-10A and breast cancer cell lines MCF-7,MDA-MB-231 and SKBR-3.MDA-MB-231 cells were selected for subsequent experi-ments,and were divided into control,si-lncRNA TTN-AS1,si-lncRNA TTN-AS1+miR-134-5p-in-hibitor and si-lncRNA TTN-AS1+EGFR-mimic groups.Cell proliferation rates were determined u-sing the CCK-8 assay.Apoptosis rates were assessed by flow cytometry.Lactic acid,glucose and a-denosine triphosphate(ATP)kits were used to detect glucose intake,lactic acid and ATP produc-tion in each group,respectively.Dual-luciferase reporter assays were performed to identify the bind-ing sites between lncRNA TTN-AS1 and miR-134-5p,as well as between miR-134-5p and EGFR.Results Compared with MCF-10A cells,the expression levels of lncRNA TTN-AS1 and EGFR mR-NA were significantly higher,while miR-134-5p expression was significantly lower in breast cancer cell lines(P＜0.05).Among these,MDA-MB-231 cells exhibited the highest expression levels of lncRNA TTN-AS1 and EGFR mRNA,and the lowest expression level of miR-134-5p(P＜0.05).Compared with the control group,the expression level of miR-134-5p was significantly increased,while the expression levels of lncRNA TTN-AS1,EGFR mRNA,and EGFR protein were significant-ly decreased in the si-lncRNA TTN-AS1 group(P＜0.05).Compared with the si-lncRNA TTN-AS1 group,the expression levels of EGFR mRNA and EGFR protein were significantly increased,and miR-134-5p expression was significantly decreased in the si-lncRNA TTN-AS1+miR-134-5p-inhibitor group and si-lncRNA TTN-AS1+EGFR-mimic group(P＜0.05).Compared with the con-trol group,cell proliferation rates were significantly reduced in the si-lncRNA TTN-AS1 group(P＜0.05).Compared with the si-lncRNA TTN-AS1 group,cell proliferation rates were significantly in-creased in the si-lncRNA TTN-AS1+miR-134-5p-inhibitor group and si-lncRNA TTN-AS1+EGFR-mimic group(P＜0.05).Compared with the control group,apoptosis rates were significantly in-creased in the si-lncRNA TTN-AS1 group(P＜0.05).Compared with the si-lncRNA TTN-AS1 group,apoptosis rates were significantly reduced in the si-lncRNA TTN-AS1+miR-134-5p-inhibitor group and si-lncRNA TTN-AS1+EGFR-mimic group(P＜0.05).Compared with the control group,glucose uptake,lactate production and ATP generation were significantly decreased in the si-lncRNA TTN-AS1 group(P＜0.05).Compared with the si-lncRNA TTN-AS1 group,glucose up-take,lactate production and ATP generation were significantly increased in the si-lncRNA TTN-AS1+miR-134-5p-inhibitor group and si-lncRNA TTN-AS1+EGFR-mimic group(P＜0.05).The du-al-luciferase reporter assay confirmed that lncRNA TTN-AS1 had a targeted binding site with miR-134-5p,and miR-134-5p has a targeted binding site with EGFR.Conclusion The lncRNA TTN-AS1 may negatively regulate the miR-134-5p/EGFR axis,thereby affecting the proliferation,apop-tosis and glycolysis of breast cancer.
