【摘要】 目的探究长链非编码RNA(LncRNA) FTX对慢性髓细胞白血病(CML)细胞伊马替尼抵抗的作用机制。方法用梯度诱导CML细胞K562伊马替尼抵抗的形成,作为抵抗组;用等体积0. 9%Na Cl处理CML细胞,作为敏感组。待抵抗组细胞处于生长对数期时,用LncRNA FTX-siRNA对其进行转染,作为转染组;用空白对照载体进行转染的作为对照组。继续培养48 h,待细胞状态稳定后,用CCK-8法检测伊马替尼的半抑制浓度(IC50),用Western Blot检测PTBP1蛋白的表达水平,用实时荧光聚合酶链反应检测LncRNA FTX和PTBP1 mRNA的表达水平。结果转染组、对照组、敏感组和抵抗组的伊马替尼IC50分别为(3. 70±0. 29),(7. 28±0. 84),(1. 41±0. 10)和(7. 06±0. 64)μmol·L-1,PTBP1蛋白分别为(0. 59±0. 02),(1. 37±0. 06),(0. 52±0. 01)和(1. 02±0. 05),LncRNA FTX分别为(0. 03±0. 01),(0. 18±0. 01),(0. 03±0. 01)和(0. 17±0. 01)。转染组的上述指标和对照组比较、敏感组的上述指标和抵抗组比较,差异均有统计学意义(均P <0. 01)。转染组和对照组的PTBP1 mRNA分别为(2. 09±0. 22)和(1. 96±0. 18),差异无统计学意义(P> 0. 05)。结论 LncRNA FTX可通过调控PTBP1蛋白的表达,从而提高CML细胞对伊马替尼的敏感性。更多还原

【Abstract】 Objective To explore the mechanism of long chain non-coding RNA( LncRNA) FTX on imatinib resistance in chronic myeloid leukemia( CML). Methods CML cells K562 imatinib resistance were induced by gradient as resistance group,and CML cells were treated with equal volume 0. 9% NaCl as sensitive group. When the cells in the resistance group were in the logarithmic phase of growth,they were transfected with LncRNA FTX-siRNA as the transfection group,and the blank control vector was used as the control group. After the cells were cultured for48 hours,the 50% inhibitory concentration( IC50) of imatinib was detected by CCK-8 method,the expression level of PTBP1 protein was detected by Western Blot,and the expression levels of LncRNA FTX and PTBP1 mRNA were detected by real-time fluorescence polymerase chain reaction. Results The IC50 of imatinib in transfection group,control group,sensitive group and resistance group were( 3. 70 ± 0. 29),( 7. 28 ± 0. 84),( 1. 41 ± 0. 10) and( 7. 06 ± 0. 64) μmol·L-1,PTBP1protein were( 0. 59 ± 0. 02),( 1. 37 ± 0. 06),( 0. 52 ± 0. 01) and( 1. 02 ± 0. 05), LncRNA FTX were( 0. 03 ± 0. 01),( 0. 18 ± 0. 01),( 0. 03 ± 0. 01) and( 0. 17 ± 0. 01). The above indexes in the transfection group were significantly different from those in the control group( all P < 0. 01). The above indexes in the sensitive group were significantly different from those in the resistance group( all P < 0. 01). The PTBP1 mRNA of transfection group and control group were( 2. 09 ± 0. 22) and( 1. 96 ± 0. 18) without significant difference( P > 0. 05). Conclusion LncRNA FTX can increase the sensitivity of CML cells to imatinib by regulating the expression of PTBP1 protein.更多还原