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。
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.
作者
王瑾
马肖容
刘海玲
姚欢
张卉
WANG Jin;MA Xiao-rong;LIU Hai-ling;YAO Huan;ZHANG Hui(Department of Hematology,The Second Affiliated Hospital of Xi’an Jiaotong University,Xi'an 710000,Shaanxi Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2019年第24期3193-3196,共4页
The Chinese Journal of Clinical Pharmacology
基金
陕西省自然科学基础研究计划基金资助项目(2016JQ8032)
