Page 46 - 南京医科大学自然版

南京医科大学学报（自然科学版） 第44卷第10期
·1362 · Journal of Nanjing Medical University（Natural Sciences） 2024年10月

·基础研究·

三氧化二砷联合普纳替尼对白血病细胞KG⁃1的抑制效应

王薇雅 ，陶长锐 ，晁红颖 ，王荣轩 ，樊 书 ，姜 玉 ，张 艳 2*
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南京医科大学附属常州市第二人民医院血液科，江苏 常州 213003；常州市第七人民医院中医科，江苏 常州 213003
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［摘 要］ 目的：探讨小分子酪氨酸激酶抑制剂普纳替尼联合三氧化二砷（arsenic trioxide，ATO）对人急性髓系白血病细胞
KG⁃1的作用及可能机制。方法：CCK⁃8法检测普纳替尼及ATO对KG⁃1细胞增殖的影响；流式细胞术 Annexin Ⅴ/PI双重染色法
检测细胞凋亡；实时荧光定量PCR检测细胞凋亡相关基因的表达；Western blot检测凋亡相关蛋白、成纤维细胞生长因子受体1
（fibroblast growth factor receptor 1，FGFR1）蛋白及信号通路分子磷酸化水平的表达变化。结果：①ATO 及普纳替尼对KG⁃1细
胞的增殖抑制作用呈剂量依赖性，两药联合较单药作用具有更高的增殖抑制率、更少的集落形成及更多的细胞凋亡，差异均有
统计学意义。②与DMSO组相比，ATO或普纳替尼均能显著下调Bcl⁃2表达，上调Bax及Caspase⁃3表达（P均<0.05）；与单药作
用相比，联合用药促进Bax及Caspase⁃3表达的作用更强（P均 < 0.01）。③普纳替尼显著抑制 FGFR1基因及蛋白的表达（P均<
0.01），ATO的加入并未使FGFR1表达进一步下降。信号通路研究显示，ATO可以显著抑制m⁃TOR和MAPK、STAT5的磷酸化
（P均<0.001），但对PI3K/AKT、STAT3的磷酸化无明显影响。普纳替尼可以显著抑制FGFR1蛋白表达及STAT3、STAT5的磷酸
化（P均<0.001），但对PI3K/AKT及MAPK的磷酸化无明显影响。两药联合后，STAT3的磷酸化水平较ATO或普纳替尼单药组
进一步下调（P均<0.01）。结论：普纳替尼及ATO可能通过不同机制抑制KG⁃1细胞增殖及集落形成并诱导细胞凋亡；两药联
合可进一步增强对KG⁃1细胞株的抑制效应。
［关键词］ 三氧化二砷；FGFR1蛋白；KG⁃1细胞；凋亡
［中图分类号］ R733.7 ［文献标志码］ A ［文章编号］ 1007⁃4368（2024）10⁃1362⁃07
doi：10.7655/NYDXBNSN220992

Effects of the arsenic trioxide and ponatinib against human leukemic KG⁃1 cells
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WANG Weiya ，TAO Changrui ，CHAO Hongying ，WANG Rongxuan ，FAN Shu ，JIANG Yu ，ZHANG Yan 2*
1 Department of Hematology，Changzhou Second Hospital of Nanjing Medical University，Changzhou 213003；
2 Department of Traditional Chinese Medicine，the Seventh People’s Hospital of Changzhou，Changzhou 213003，China
［Abstract］ Objective：To explore the effects and possible mechanisms of the arsenic trioxide（ATO）and small molecule tyrosine
kinase inhibitor ponatinb on KG⁃1 cells in vitro. Methods：Effects of ATO and ponatinib on proliferation of KG⁃1 cells were detected by
CCK⁃8，and the apoptosis was assessed by Annexin V⁃FITC. Reverse transcription quantitative polymerase chain reaction（q⁃PCR）
analysis was used to detect the expression of apoptosis⁃related genes. Western blott was performed to explore the expression levels of
apoptosis⁃related proteins，fibroblast growth factor receptor 1（FGFR1）and phosphorylated signal molecules. Results：①Both ATO and
ponatinib effectively inhibited cell proliferation by dose dependent manners. The combination of the two drugs exhibited higher
proliferation inhibition rate，less colony formation and more cell apoptosis compared to the single drug treatment. ②Compared with the
DMSO group，treatment with either ATO or ponatinib led to significant down⁃regulation of Bcl⁃2，up⁃regulation of Bax and Caspase⁃3
（P < 0.05）. The combination of the two drugs up⁃regulated the expression of Bax and Caspase⁃3 more than single drug treatment（both
P < 0.01）. ③Punatinib significantly inhibited the expression of FGFR1 gene and protein（both P < 0.01），and the addition of ATO did
not decrease FGFR1 expression further. Signaling pathway studies showed that ATO significantly inhibited the phosphorylation of
MAPK，m ⁃ TOR，and STAT5，but had no significant effect on the phosphorylation of PI3K/AKT and STAT3. Ponatinib markedly
inhibited the phosphorylation of STAT3/5，and FGFR1 expression（both P < 0.001），but had no significant effect on the phosphorylation
of PI3K/AKT and MAPK. The phosphorylation level of STAT3 was further down⁃regulated by the combination of the two drugs compared

［基金项目］ 常州市科协软科学研究课题
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通信作者（Corresponding author），E⁃mail：3390198019@qq.com