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Archive > Volume 29 Issue 6 > 2022,29(6):541-548. DOI:10.3872/j.issn.1007-385X.2022.06.005 Prev Next
Basic Research
miR-323a-3p regulats the proliferation, migration and invasion of NSCLC A549 cells through targeting TM4SF1
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Abstract:
Objective: To explore the expression of miR-323a-3p and transmembrane 4 super family member 1 (TM4SF1) in lung cancer tissues and cells, and the target regulatory relationship between them, and to observe their effects on the proliferation, migration and invasion of lung cancer A549 cells, as well as their effects on the growth of A549 cell transplanted tumors in nude mice. Methods:Twenty pairs of lung cancer tissues and their corresponding paracancerous tissues were collected from January 2014 to December 2014 in Qinghai Provincial People's Hospital. The expressions of miR-323a-3p, mRNA and TM4SF1 expression of TM4SF1 in lung cancer tissues were detected by qPCR and WB methods. A549 cells were transfected with miR-323a-3p mimic, and the effects of miR-323a-3p overexpression on cell proliferation, migration and invasion as well as protein expression of TM4SF1, cyclin D1, p21, MMP-2 and MMP-9 were detected by MTT method, Transwell method and WB method, respectively. The targeting relationship between miR-323a3p and TM4SF1 was analyzed using the bioinformatics prediction tool StarBase and dual-luciferase reporter gene experiments. A549 cells were transfected with si-TM4SF1, miR-323a-3p+pcDNA or miR-323a-3p+pcDNA-TM4SF1, respectively, and the changes in proliferation, migration and invasion of A549 cells were evaluated. In the meanwhile, xenograft model was established in BALB/c nude mice by transplanting A549 cells with various treatment, and the xenograft volume was measured and calculated on day 14, 21 and 28.Results: Compared with paracancerous tissues, the expression levels of miR-323a-3p in lung cancer tissues were significantly downregulated, while the mRNA and protein levels of TM4SF1 were significantly up-regulated (all P<0.01). Overexpression of miR-323a-3por inhibition of TM4SF1 expression reduced the proliferation, migration, and invasion of A549 cells, as well as the expression of cyclin D1, MMP-2, and MMP-9 proteins, but promoted the expression of p21 protein (all P<0.01). StarBase prediction and dual-luciferase gene reporter experiment showed that miR-323a-3p could complimentarily bind with TM4SF1 and regulate its expression. After upregulation of TM4SF1 expression, the inhibitory effects of miR-323a-3p overexpression on malignant biological behaviors of A549 cells and protein expression of cyclin D1, MMP-2 and MMP-9, as well as tumor growth in nude mice were reversed (all P<0.01);moreover, the promotive effect of miR-323a-3p overexpression on the protein expression of p21 was also reversed (P<0.01).Conclusion: miR-323a-3p inhibits proliferation, migration and invasion of A549 cells and suppresses xenograft growth in nude mice by downregulating TM4SF1 expression.
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