LncRNA LHFPL3-AS1 Regulates the Proliferation and Invasion of Keloid Fibroblasts by Targeting the miR-145-5p/ATF3 Axis
WU Xixi1, ZHOU Chuchao1, WANG Jingjing2, YANG Yanqing1*
Keloids are pathological scars formed after skin lesions. This study aims to explore the effect of LncRNA LHFPL3-AS1 targeting miR-145-5p/ATF3 axis on the proliferation and invasion of keloid fibroblasts. The keloid fibroblast line HKF was assigned into control group, si-NC group, si-LHFPL3-AS1 group, si-LHFPL3-AS1+inhibitor NC group, si-LHFPL3-AS1+miR-145-5p inhibitor group, miR-NC group, miR-145-5p mimic group, miR-145-5p mimic+pc-NC group, and miR-145-5p mimic+pc-ATF3 group. qRT-PCR was used to detect the mRNA expression levels of LncRNA LHFPL3-AS1, miR-145-5p, and ATF3. MTT assay was used to detect cell prolifera tion. Transwell method was used to detect invasion. Flow cytometry was used to detect cell apoptosis. Western blot was used to detect ATF3 and Collagen I proteins. Dual luciferase reporter gene assay was used to detect the interaction between LncRNA LHFPL3-AS1 and miR-145-5p, as well as miR-145-5p and ATF3. The results showed that com pared with normal skin cells, the levels of LncRNA LHFPL3-AS1 and ATF3 mRNA in HKF cells raised, while the level of miR-145-5p reduced (P<0.05). Compared with si-NC group, the LncRNA LHFPL3-AS1 level, ATF3 protein expression, cell proliferation and invasion abilities in HKF cells in si-LHFPL3-AS1 group were all reduced, and the miR-145-5p level and cell apoptosis rate were increased (P<0.05). Compared with si-LHFPL3-AS1+inhibitor NC group, the miR-145-5p level of HKF cells in the si-LHFPL3-AS1+miR-145-5p inhibitor group was decreased, the ATF3 protein expression, cell proliferation and invasion abilities were improved, and the apoptosis rate was decreased (P<0.05). Compared with miR-NC group, miR-145-5p mimic group showed a decrease in ATF3 protein expression, reduced cell proliferation and invasion abilities, and raised apoptosis rate in HKF cells (P<0.05). Compared with miR-145-5p mimic+pc-NC group, miR-145-5p mimic+pc-ATF3 group showed raised expression of ATF3 protein, enhanced proliferation and invasion abilities, and reduced apoptosis rate in HKF cells (P<0.05). Dual luciferase activ ity showed that LncRNA LHFPL3-AS1 had a targeted relationship with miR-145-5p, and miR-145-5p had a targeted relationship with ATF3 (P<0.05). In summary, LncRNA LHFPL3-AS1 may regulate proliferation and invasion of HKF cells by targeting miR-145-5p/ATF3 axis.
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