长链非编码RNA TTTY15在骨肉瘤组织中的表达及其对骨肉瘤细胞活力和侵袭的影响

Long non-coding RNA TTTY15 expression in osteosarcoma and its effect on viability and invasion ability of osteosarcoma cells

目的:观察长链非编码RNA TTTY15在骨肉瘤组织及细胞株中的表达,并探讨其对骨肉瘤细胞活力和侵袭的影响。方法:应用qPCR法检测11例骨肉瘤组织及其瘤旁组织中TTTY15的水平,同时检测TTTY15在骨肉瘤细胞株(143B、Saos2、MG-63、U2OS和HOS)和人成骨细胞株hFOB1.19中的表达。在表达量最高的细胞株(MG-63)中通过转染小干扰RNA敲减TTTY15的表达,CCK-8法检测TTTY15低表达对细胞活力的影响,流式细胞术检测其对细胞周期的影响,Transwell检测其对细胞侵袭能力的影响,qPCR检测miR-216b-5p和FOXM1 mRNA的表达,Western blot法检测相关蛋白的变化。结果 :与瘤旁正常组织相比,TTTY15在骨肉瘤组织中表达升高(P<0.01);与人成骨细胞相比,TTTY15在骨肉瘤细胞株中表达升高(P<0.05),其中在MG-63细胞中表达水平最高(P<0.01)。敲减TTTY15在MG-63细胞的表达后,细胞活力降低(P<0.05),细胞周期被抑制在G_0/G_1期(P<0.01),细胞的侵袭能力降低(P<0.01),miR-216b-5p mRNA表达水平升高(P<0.01),FOXM1的mRNA表达降低(P<0.01),同时FOXM1、CDK4、cyclin D1、MMP-2和N-cadherin的蛋白表达降低,而E-cadherin的蛋白表达升高(P<0.05)。结论:TTTY15在骨肉瘤组织和细胞株中表达增高,敲减TTTY15的表达可抑制骨肉瘤细胞的活力和侵袭能力,其机制可能与TTTY15低表达导致miR-216b-5p的表达升高,引起FOXM1基因表达下调有关。

AIM： To observe the expression of long noncoding RNA TTTY15 in osteosarcoma tissues and cell lines and to explore its effect on the viability and invasion ability of osteosarcoma cell lines. METHODS： qPCR was used to detect the expression of TTTY15 in 11 cases of osteosarcoma and its adjacent tissues. The mRNA levels of TTTY15 in osteosarcoma cell lines（ 143 B,Saos2,MG-63,U2OS and HOS） and human osteoblast cell line hFOB1. 19 were also tested.TTTY15 was down-regulated after transfected with small interfering RNA in MG-63 cells,the cell line with the highest level of TTTY15. The effect of TTTY15 knockdown on the viability of MG-63 cells was measured by CCK-8 assay. The cell cycle distribution was analyzed by flow cytometry. The effect of TTTY15 knockdown on the cell invasion ability was detected by Transwell assay. The levels of miR-216b-5p and FOXM1 mRNA were detected by q PCR,and the changes of the related proteins were determined by Western blot. RESULTS： Compared with the adjacent tissues,the expression of TTTY15 increased in the osteosarcoma tissues（ P〈0. 01）. Compared with the human osteoblast cell line,the expression of TTTY15 increased in the osteosarcoma cell lines（ P〈0. 05）,and the level of TTTY15 in the MG-63 cells was the highest（ P〈0. 01）. After knockdown of TTTY15 expression in the MG-63 cells,the cell viability was decreased（ P〈0. 05）,cell cycle progression was inhibited（ P〈0. 01）,and the cell invasion ability was decreased（ P〈0. 01）. The expression of miR-216 b-5 p was increased（ P〈0. 01） and the expression of FOXM1 mRNA was decreased（ P〈0. 01）. The protein expression of FOXM1,CDK4,cyclin D1,MMP-2 and N-cadherin was decreased,while the protein expression of E-cadherin was increased（ P〈0. 05）. CONCLUSION： The expression of TTTY15 is increased in the osteosarcoma tissues and cell lines. The low expression of TTTY15 inhibits the cell viability and invasion ability of osteosarcoma cells. The possible mechanism is that the knockdown of TTTY15 expression results in the increase in miR-216b-5p expression and the downregulation of FOXM1 expression.