Effects of LncRNA ASB16-AS1 on the Proliferation, Migration, and Invasion of Oral Squamous Cell Carcinoma Cells by Regulating the miR-185-5p/TRAM2 Axis
JIN Rui1, JIAO Hezhu2, HUANG Ying2, ZHANG Min2*
This study aims to investigate the effects of LncRNA ASB16-AS1 (long non-coding RNA ASB16-AS1) on the proliferation, migration, and invasion of oral squamous cell carcinoma cells by targeting the miR-185-5p/TRAM2 axis. Human oral squamous cell carcinoma cells SCC-9 were cultured in vitro and grouped into control group, sh-NC group, sh-ASB16-AS1 group, sh-ASB16-AS1+inhibitor NC group, and sh-ASB16-AS1+miR-185-5p inhibitor group. qRT-PCR method was applied to detect the expression of LncRNA ASB16-AS1, miR-185-5p, and TRAM2 mRNA of cells in each group. SCC-9 cell proliferation was detected using CCK-8 assay kit. SCC-9 cell apoptosis was detected by flow cytometry. SCC-9 cell migration was detected using scratch assay, and SCC-9 cell invasion was detected using Transwell method. Western blot was applied to detect the expression of TRAM2 protein. Dual luciferase reporter assay was applied to verify the targeting relationship between LncRNA ASB16-AS1 and miR-185-5p, and miR-185-5p and TRAM2. Compared with the control group and sh-NC group, the expression levels of LncRNA ASB16-AS1, TRAM2 mRNA, and TRAM2 protein, as well as D value, scratch healing rate, and number of invasive cells were reduced in the sh-ASB16-AS1 group, while the level of miR-185-5p and cell apoptosis rate were increased (P<0.05). Compared with the sh-ASB16-AS1+inhibitor NC group, the miR-185-5p level and cell apoptosis rate in the sh-ASB16-AS1+miR-185-5p inhibitor group were reduced, while the expression levels of TRAM2 mRNA and TRAM2 protein, D values, scratch healing rate, and number of invasive cells were elevated (P<0.05), however, there were no significant changes in the levels of LncRNA ASB16-AS1 (P>0.05). Biological information websites predicted that miR-185-5p had targeted binding sites with LncRNA ASB16-AS1 and TRAM2, and dual luciferase reporter gene experiments confirmed the existence of targeted relationships between LncRNA ASB16-AS1 and miR-185-5p, and miR-185-5p and TRAM2 (P<0.05). The expression of LncRNA ASB16-AS1 is upregulated in oral squamous cell carcinoma cells. Inhibiting the expression of LncRNA ASB16-AS1 can target and upregulate the expression of miR-185-5p, inhibit the expression of TRAM2, promote apoptosis of oral squamous cell carcinoma cells, and inhibit their proliferation, migration, and invasion.
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