Study On The Role Of Three LncRNAs In Melanoma Cells

Objective In vitro studies of LncRNAs(RP11-875O11.3-001,RP11-496I2.2,RP11-379K17.12-001)in human cutaneous malignant melanoma cells A375,A875 and normal human epidermal melanocyte HEM-M cells Expression,and its effect on melanoma cell proliferation,invasion,cycle,apoptosis.Methods Human melanoma cell line A375,A875 and normal human epidermal melanocyte HEM-M were cultured in vitro.The expression levels of LncRNA RP11-875O11.3-001,LncRNA RP11-496I2.2 and LncRNA RP11-379K17.12-001 in A375,A875 and HEM-M were detected by real-time quantitative PCR(qRT-PCR);A375,A875 and HEM-M cells were transfected into corresponding siRNAs,respectively,to reduce the expression level of target LncRNA(divided into experimental group and control group,in which the experimental group was transfected with the target gene siRNA;the control group was treated with negative control siRNA).After plasmid transfection,the absorbance of cells at 450 nm was detected by CCK8 assay(24h,48 h,72h)to reflect cell proliferation;Transwell chamber assay was used to detect cell invasion ability;flow cytometry was used to determine cell cycle and apoptosis rate.Results Compared with HEM-M,LncRNA RP11-875O11.3-001,LncRNA RP11-496I2.2,LncRNA RP11-379K17.12-001 were highly expressed in A375 and A875,the difference was statistically significant(P<0.05).The expression of RP11-875O11.3-001,RP11-496I2.2 and RP11-379K17.12-001 was knocked down by plasmid transfection.The expression levels of three LncRNAs in the experimental group were significantly lower than those in the control group.Statistically significant(P<0.001).The results of CCK8 showed that in the A375 cells,the OD450 of the three LncRNAs in the experimental group was lower than that in the control group(P<0.05).In the A875 cells,the OD450 of the three LncRNAs in the experimental group was lower than that in the control group at 72 h(P<0.05).In the HEM-M cells,the OD values of the three LncRNAs in the experimental group were compared with the control group.There was no difference(P>0.05).The results of Transwell chamber showed that the expression of three LncRNAs was knocked down.The number of transmembrane cells in the experimental group of melanoma cells A375 and A875 was lower than that of the control group,and the results were statistically significant(P<0.05).The results of flow cytometry showed that compared with the control group,the G0/G1 phase cells of A375 and A875 cells in the experimental group increased(P<0.05),and the S phase decreased(P<0.05).The results of apoptosis experiment showed that the apoptosis rate of A375 and A875 cells in the experimental group was higher than that in the control group,and the difference was statistically significant(P<0.05).The apoptosis rate of HEM-M had no effect(P>0.05).Conclusions RP11-875O11.3-001,RP11-496I2.2,RP11-379K17.12-001 were highly expressed in melanoma cell lines A375 and A875;CCK8 experiments showed that knocking down the expression level of LncRNAs inhibited the proliferation of melanoma cells A375 and A875,and had no effect on the proliferation of human normal epidermal melanocytes HEM-M.Transwell and flow cytometry experiments showed that down-regulation of LncRNA RP11-875O11.3-001,LncRNA RP11-496I2.2,LncRNA RP11-379K17.12-001 expression levels inhibited the invasion of melanoma cells and induced melanin Tumor cell apoptosis,while causing melanoma cell growth arrest in G0/G1 phase.LncRNA RP11-875O11.3-001,LncRNA RP11-496I2.2,LncRNA RP11-379K17.12-001 may be involved in the malignant progression of melanoma.The results of this study show the important role of LncRNAs in the development of melanoma,and may Become a new target for melanoma treatment.