长链非编码RNA SNHG15对肾细胞癌细胞增殖、凋亡、迁移和侵袭的影响

Effects of LncRNA SNHG15 on proliferation,migration and invasion of RCC cells

目的:探讨长链非编码RNA(LncRNA)SNHG15对肾细胞癌(RCC)细胞增殖、凋亡、迁移和侵袭的影响及作用机制。方法:786-O和ACHN细胞分为转染空载体pcDNA3.1的空白对照组和转染SNHG15表达质粒的SNHG15组;分为转染无关序列的shRNA阴性对照组(shNC组)、转染靶向SNHG15的短发夹RNA组1和组2(shSNHG15#1组和shSNHG15#2组)。分别采用MTT法、Caspase 3/7活性试剂盒、划痕实验和Transwell小室检测786-O和ACHN细胞的增殖、凋亡、迁移和侵袭能力。生物信息学预测和双荧光素酶报告实验验证SNHG15、miR-665和Myc之间的关系。结果:SNHG15过表达可增加786-O和ACHN细胞SNHG15的表达,而短发夹RNA敲低SNHG15的表达(P<0.001)。SNHG15过表达可促进786-O和ACHN细胞的增殖、凋亡抵抗、迁移和侵袭(P<0.05),敲低SNHG15表达可抑制786-O和ACHN细胞的以上恶性生物学行为(P<0.05)。双荧光素酶报告实验证实SNHG15可直接结合miRNA-665,解除miR-665对其靶蛋白Myc的抑制,增加Myc蛋白表达。结论:SNHG15可促进RCC细胞的增殖、凋亡抵抗、迁移和侵袭,可能与通过竞争性吸附miR-665进而增加Myc蛋白表达有关。

Aim:To investigate the effects and mechanism of LncRNA SNHG15 on proliferation,apoptosis,migration,and invasion of renal cell carcinoma(RCC) cells.Method:786-O and ACHN cells were divided into the blank control group transfected with the empty vector pcDNA3.1 and the SNHG15 group transfected with the SNHG15 expression plasmid;divided into shRNA negative control group transfected with unrelated sequences(shNC group),short hairpin RNA group 1 and group 2 transfected short hairpin RNA targeting SNHG15(shSNHG15#1 group and shSNHG15#2 group).MTT assay,Caspase 3/7 kit,scratch assay,and Transwell chamber were used to detect the proliferation,apoptosis,migration,and invasion ability of RCC cells.The relationship between SNHG15,Myc,and miR-665 was detected through bioinformatics prediction and dual luciferase reporter assay.Results:Overexpression of SNHG15 significantly increased the expression of SNHG15 in RCC cells,while short hairpin RNA transfection inhibited its expression(P<0.001).Overexpression of SNHG15 promoted the proliferation,apoptosis resistance,migration,and invasion abilities of 786-O and ACHN cells(P<0.05),while knocking down SNHG15 inhibited the above malignant biological behavior of RCC cells(P<0.05).The dual luciferase reporter assay confirmed that SNHG15 could directly bind to miRNA-665,thereby relieving the inhibition of miR-665 on its target protein Myc and increasing the expression of Myc protein.Conclusion:SNHG15 may promote the proliferation,apoptosis resistance,migration,and invasion abilities of RCC cells through increasing the expression of Myc protein by competitive adsorption of miR-665.