lncRNA HIF1A-AS1对长春新碱耐药视网膜母细胞瘤细胞化疗敏感性的影响

Effect of lncRNA HIF1A-AS1 on chemotherapy sensitivity of vincristine-resistant in retinoblastoma cells

摘要:

目的：探讨长链非编码RNA-HIF1A-AS1(lncRNA HIF1A-AS1)调节缺氧诱导因子-1α(HIF-1α))表达对长春新碱(VCR)耐药视网膜母细胞瘤(RB)细胞化疗敏感性的影响。

方法：建立人RB VCR耐药细胞株SO-RB50/VCR，实时荧光定量PCR(RT-qPCR)检测SO-RB50与SO-RB50/VCR细胞lncRNA HIF1A-AS1表达； 在SO-RB50/VCR细胞中抑制lncRNA HIF1A-AS1表达或同时过表达HIF-1α，检测SO-RB50/VCR细胞对VCR的半数抑制浓度(IC₅₀)及细胞增殖、凋亡情况； Western blot检测HIF-1α、多药耐药相关蛋白(MRP)、P-糖蛋白(P-gp)蛋白表达。

结果：与SO-RB50细胞相比，SO-RB50/VCR细胞中lncRNA HIF1A-AS1与HIF-1α蛋白表达水平升高(P<0.05)； 在SO-RB50/VCR细胞中抑制lncRNA HIF1A-AS1表达后，细胞凋亡率显著升高(P<0.05)，细胞吸光度(OD₄₅₀)值显著降低，VCR对细胞的IC₅₀值及HIF-1α、MRP、P-gp蛋白表达水平显著降低(P<0.05)； 过表达HIF-1α可减弱下调lncRNA HIF1A-AS1表达对SO-RB50/VCR细胞耐药性的抑制作用。

结论：lncRNA HIF1A-AS1在SO-RB50/VCR细胞中高表达，抑制lncRNA HIF1A-AS1表达可通过下调HIF-1α表达，降低SO-RB50/VCR细胞对VCR的耐药性。

Abstract:

AIM: To investigate the effect of long non-coding RNA-HIF1A-AS1(lncRNA HIF1A-AS1)on the chemotherapy sensitivity of vincristine(VCR)-resistant in retinoblastoma(RB)cells by regulating the expression of hypoxia-inducible factor-1α(HIF-1α).

METHODS: The human RB VCR-resistant cell line SO-RB50/VCR was established, expression of lncRNA HIF1A-AS1 in SO-RB50 and SO-RB50/VCR cells were detected by reverse transcription-quantitative real-time PCR(RT-qPCR); inhibition of lncRNA HIF1A-AS1 expression or simultaneous overexpression of HIF-1α in SO-RB50/VCR cells, and then median inhibitory concentration(IC₅₀)of VCR and cell proliferation and apoptosis were detected in SO-RB50/VCR cells; the protein expressions of HIF-1α, multidrug resistance associate protein(MRP)and P-glycoprotein(P-gp)were measured by Western blot.

RESULTS: Compared with SO-RB50 cells, the expression levels of lncRNA HIF1A-AS1 and HIF-1α protein in SO-RB50/VCR cells were increased(P<0.05); after inhibiting the expression of lncRNA HIF1A-AS1 in SO-RB50/VCR cells, the apoptosis rate was significantly increased(P<0.05), optical density(OD₄₅₀), the IC₅₀ value of VCR on cells and the expression levels of HIF-1α, MRP and P-gp proteins were significantly reduced(P<0.05); overexpression of HIF-1α attenuates the inhibitory effect of down-regulated lncRNA HIF1A-AS1 expression on drug resistance in SO-RB50/VCR cells.

CONCLUSION: The lncRNA HIF1A-AS1 was highly expressed in SO-RB50/VCR cells, and inhibition of lncRNA HIF1A-AS1 expression reduced VCR resistance in SO-RB50/VCR cells by down-regulating HIF-1α expression.