Preliminary screening and correlation analysis for lncRNAs related to radiosensitivity in melanoma cells by inhibiting glycolysis. / ç³–é µè§£å‚ä¸Žçš„é»‘è‰²ç´ ç˜¤ç»†èƒžæ”¾å°„æ•æ„Ÿæ€§lncRNAsçš„åˆæ­¥ç­›é€‰åŠç›¸å ³æ€§åˆ†æž.

ABSTRACT

OBJECTIVES: To screen the expression profiles of lncRNA and mRNA related to the radiosensitivity of melanoma cells by inhibiting glycolysis through microarray technology. METHODS: WM35 melanoma cells were treated with different concentrations (1.25, 2.50, 5.00, 10.00 mmol/L) of 2-deoxy-D-glucose (2-DG) and different doses (0, 2, 4, 6, 8 Gy) of X-ray irradiation. MTT assay was used to detect the proliferation ability of NC-0 Gy group (negative control group), NC-4 Gy group (only 4 Gy X-ray irradiation), 2-DG group (only 2.50 mmol/L DG treatment), and 2-DG-4 Gy group (2.50 mmol/L 2-DG treatment, 4 Gy X-ray irradiation). Microarray chip was used to detect the changes in the expression profiles of lncRNA and mRNA in the NC-4 Gy group and the 2-DG-4 Gy group. Real-time RT-PCR was used to quantitatively detect the top 5 upregulated and the top 5 downregulated expression lncRNA. CNC analysis was used to predict potential target genes for the 10 most significantly expressed lncRNAs, after which the co-expression network of lncRNA and co-regulated mRNA were constructed. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were used to predict the functional distribution of differentially expressed lncRNA. Real-time RT-PCR was used to quantitatively detect the top 5 upregulated and the top 5 downregulated expression lncRNAs. RESULTS: After 48 and 96 h, the cell proliferation of WM35 treated with 2-DG was significantly inhibited in a dose-dependent manner (all P<0.05). The cell proliferation of WM35 was inhibited by a high dose of X-ray irradiation, resulting in the death of mass cells. The cell proliferation activity of WM35 after 4 Gy X-ray irradiation descended 61% compared to the negative control group. Microarray analysis showed that there were 1 206 lncRNAs and 543 differentially expressed mRNAs between the NC-4 Gy group and the 2-DG-4 Gy group, while real-time RT-PCR showed basically consistent changes in lncRNA and mRNA microarray. Further CNC analysis showed that these 10 lncRNAs had a positive or negative correlation with 333 target genes. GO analysis was mainly concentrated in DNA binding, DNA damage repair, cell cycle arrest, and oxidative stress, while KEGG pathway analysis showed the 10 lncRNAs were related to radiosensitivity. CONCLUSIONS: Microarray chip screens the expression profiles of differentially expressed lncRNA related to the radiosensitivity of melanoma cells via inhibiting glycolysis, and lncRNA RPL34-AS1 might be a potential biological target for melanoma radiotherapy.