ABSTRACT
Objective:
To investigate the expression of
long non-coding RNA (
lncRNA) MTATP6P1 in
melanoma and its effect on the proliferation, migration and invasion of
melanoma cells by targeting
miRNA-411-5p (miR-411-5p).
Methods:
A total of 461 samples of
melanoma tissues and paracancerous
tissues (>2 cm from the
tumor margin) were collected from the
tumor-associated
lncRNA database (TANRIC database updated in July 2021), and the expression of MTATP6P1 was compared between the two groups. The
bioinformatics software lncRNA Disease v2.0 was used to predict the possible
binding site microRNA (
miRNA) of MTATP6P1. Real-
time fluorescent quantitative
polymerase chain reaction (qRT-
PCR) was used to detect the relative expression level of MTATP6P1 in
melanoma cells A-375, WM266-4, VMM5A, A2058 and normal
human epidermal
melanocytes PIG1; and the lowest relative expression level of
cells in MTATP6P1 were divided into MTATP6P1 group (transfected with MTATP6P1 overexpression
plasmid) and NC group (transfected with blank
plasmid). The proliferation
ability of A-375
cells was detected by using
CCK-8 method; the migration
ability of A-375
cells was detected by using scratch test; the invasion
ability of A-375
cells was detected by using Transwell assay; the targeting relationship between MTATP6P1 and miR-411-5p was detected by using dual
luciferase reporter gene assay;
Western blot was used to detect the expression of ERK signaling pathway related
proteins in
cells.
Results:
The relative expression levels of MTATP6P1 in
melanoma tissues and adjacent
tissues were 9.82±0.58 and 11.56±0.16, respectively. The expression level of MTATP6P1 in
melanoma tissues was lower than that in paracancerous
tissues ( t = 9.56, P = 0.009). The relative expression levels of MTATP6P1 in normal
human epidermal
melanocyte PIG1 and
melanoma cells A-375, WM266-4, VMM5A, and A2058 were 1.01±0.13, 0.12±0.02, 0.66±0.04, 0.39±0.07, 0.49±0.05; the relative expression level of MTATP6P1 in
melanoma cells was lower than that in PIG1
cells (all P < 0.05), and then A-375
cells with the lowest relative expression level were taken to perform the subsequent experiments. The relative expression levels of MTATP6P1 in A-375
cells of MTATP6P1 group and NC group were 14.83±1.67 and 1.02±0.30, respectively ( t = 8.13, P < 0.001). After 16, 24, 32, and 40 h of
culture, the proliferation
ability of the
cells in the MTATP6P1 group was lower than that in NC group (all P < 0.05). The scratch healing rates of A-375
cells in MTATP6P1 group and NC group were (26±7)% and (55±4)%, respectively; the scratch healing rate in MTATP6P1 group was lower than that in NC group ( t = 3.48, P = 0.009). The invasive number of A-375
cells in MTATP6P1 group and NC group were (32±12) and (116±17), respectively; the number of invasive
cells in MTATP6P1 group was lower than that in NC group ( t = 4.11, P = 0.006). The results of dual
luciferase reporter gene assay showed that there was a targeting relationship between MTATP6P1 and miR-411-5p. The relative expression level of miR-411-5p in A-375
cells of MTATP6P1 group and NC group was 1.04±0.16 and 5.37±0.68, respectively; the expression level of miR-411-5p in MTATP6P1 group was lower than that in NC group ( t = 6.20, P < 0.001). The expressions of ERK signaling pathway
proteins p-
Ras, p-Raf, p-MEK1, p-RSK, and
AP-1 in A-375
cells of MTATP6P1 group were lower than those in NC group.
Conclusions:
MTATP6P1 inhibits the proliferation, migration and invasion of
melanoma A-375
cells through targeting miR-411-5p.
