Expression of long non-coding RNA MTATP6P1 in melanoma and its effect on melanoma cell proliferation, migration and invasion by targeting miRNA-411-5p
10.3760/cma.j.cn115355-20230203-00058
- VernacularTitle:长链非编码RNA MTATP6P1在黑色素瘤中的表达及其靶向miRNA-411-5p对黑色素瘤细胞增殖、迁移和侵袭的影响
- Author:
Yaqiong WU
1
;
Haixia JING
;
Yao YANG
;
Lingling WANG
;
Lihua WANG
;
Yanqing HE
Author Information
1. 十堰市太和医院（湖北医药学院附属医院）皮肤科，十堰 442000
- Keywords:
Melanoma;
Long non-coding RNA;
MicroRNAs;
Cell proliferation;
Cell migration assays;
Neoplasm invasiveness
- From:
Cancer Research and Clinic
2023;35(12):881-886
- CountryChina
- Language:Chinese
-
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.
