Effect of LINC00324 on proliferation,migration and invasion of melanoma cell and its molecular mechanism
Objective To investigate the influence and its molecular mechanism of LINC00324 on the proliferation,migration and invasion abilities of melanoma cells.Methods The expression level of LINC00324 in melanoma tissues and paracancerous tissues,and its effect on patient prognosis were analyzed using gene expression profiling interactive analysis(GEPIA).The melenoma SK-MEL-2 cells were divided into four groups:negative control(NC)group,overexpression(OE)group(transfected with overexpression vector),silent negative control(si-NC)group and si-LINC00324 group(transfected with LINC00324 siRNA).The transfection efficiency of LINC00324 and the mRNA expression levels of Beclinl,LC3A/B,and Bcl-2 were determined using real-time fluorescence quantitative PCR(qRT-PCR).CCK-8 was performed to detect the cell proliferation ability.Scratch assay was used to detect the cell migration ability.Transwell assay was used to detect the cell invasion ability.The expression levels of key proteins were detected using Western blot.Results Compared with paracancerous tissues,the expression of LINC00324 was decreased in melanoma tissues(P＜0.05),and the patients with low expression had a poor prognosis.The qRT-PCR results revealed that the LINC00324 overexpression vetor and siRNA were successfully transfected into melanoma cells(P＜0.01).Compared with NC group,the proliferation,migration,and invasion abilities of SK-MEL-2 cell were significantly decreased in OE group(P＜0.01).Compared with si-NC group,the proliferation and migration abilities of SK-MEL-2 cell were increased in si-LINC00324 group(P＜0.01).Compared with NC group,the mRNA levels of Beclin1,LC3A/B,and Bcl-2 were reduced in OE group(P＜0.01).Compared with si-NC group,the mRNA levels of Beclin1,LC3A/B,and Bcl-2 were increased in si-LINC00324 group(P＜0.01).Compared with NC group,the protein levels of p62 and Bcl-2 were upregulated(P＜0.05),while the protein level of LC3B Ⅱ was downregulated in OE group(P＜0.01).Compared with si-NC group,the protein levels of p62 and Bcl-2 were downregulated(P＜0.05),while the protein level of LC3BⅡ was upregulated in si-LINC00324 group(P＜0.05).Conclusion The expression of LINC00324 is low in melanoma tissues and cells,which can affect melanoma cell proliferation,migration,and invasion by suppressing autophagy via Bcl-2 upregulation.
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