线粒体铁蛋白调控上皮-间质转化对非小细胞肺癌细胞顺铂耐药的影响
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1.吉林医药学院公共卫生学院,吉林 吉林 132013; 2.延边大学基础医学院,吉林 延边 133002

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R-33

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Cisplatin resistance in non-small lung cancer cells is affected by ferritin mitochondrial through EMT regulation
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1. School of Public Health, Jilin Medical University, Jilin 132013, China. 2. School of Basic Medicine, Yanbian University, Yanbian 133002

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    摘要:

    目的 研究线粒体铁蛋白(FtMt)对非小细胞肺癌细胞顺铂耐药株 A549 / DDP 和非小细胞肺癌细胞 株 A549 上皮-间质转化(EMT)的影响,探讨其改善顺铂耐药的分子机制。 方法 流式细胞术检测 A549 和 A549 / DDP 的凋亡率和细胞周期情况,Western blot 检测 E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和 FtMt 蛋白 表达水平, RT-qPCR 法检测细胞中 E-cadherin、 N-cadherin、 snail、 slug、 twist、 波形蛋白 ( vimentin)、 铁转运蛋白 (ferroportin,FPn)、二价金属离子转运蛋白 1 ( divalent metal transporter 1, DMT1)、 铁调素和转铁蛋白受体 1 (transferrin receptor1,TfR1)基因表达水平。细胞划痕实验检测细胞迁移情况。 结果 随着顺铂使用剂量增加, A549 / DDP 和 A549 细胞凋亡率逐渐升高,且 A549 细胞凋亡率显著高于 A549 / DDP 细胞。 顺铂处理后,与 0 μg / mL 顺铂组比较,顺铂各剂量组 A549 细胞 G0 / G1 期和 G2 / M 期逐渐降低,A549 / DDP 细胞 G0 / G1 期和 G2 / M 期先升高后降低(P<0. 05)。 A549 / DDP 细胞 E-cadherin 和 mRNA 水平低于 A549 细胞,A549 / DDP 细胞 N-cadherin 和 mRNA 水平高于 A549 细胞,A549 / DDP 细胞中 snial、slug、vimentin、FPn、铁调素( hepcidin)和 TfR1 mRNA 水平高于 A549 细胞(P<0. 05),A549 / DDP 细胞中 DMT1 mRNA 水平低于 A549 细胞(P>0. 05),A549 / DDP 细胞迁移的速度显著快于 A549 细胞(P<0. 05),A549 / DDP 细胞中 FtMt 蛋白表达水平显著高于 A549 细胞(P<0. 05)。 结论 肺癌细胞的顺铂耐药与 EMT 过程有关,FtMt 在肺癌细胞耐受顺铂过程中可能发挥了重要作用。

    Abstract:

    Objective To investigate the effects of mitochondrial ferritin (FtMt) on epithelial mesenchymal transition (EMT) of-cisplatin-resistant non-small lung cancer line A549 / DDP and non-small lung cancer line A549, and to explore the molecular mechanism in improving cisplatin resistance. Methods Flow cytometry was used to determine the apoptosis rate and cell cycle of A549 and A549 / DDP cells. Western blot was used to analyze expression of E-cadherin, N- cadherin and FtMt. RT-qPCR was used to measure gene expression of E-cadherin, N-cadherin, snail, slug, twist, vimentin, FPn, DMT1, hepcidin and TfR1 in cells. Cell migration was assessed by a scratch assay. Results The apoptotic rate of A549 / DDP and A549 cells was increased with the increase in cisplatin concentration, and the apoptotic rate of A549 cells was significantly higher than that of A549 / DDP cells. After cisplatin treatment, A549 cells in G0 / G1 and G2 / M phases were gradually decreased in all groups compared with the 0 μg / mL cisplatin group, whereas A549 / DDP cells in G0 / G1 and G2 / M phases were first increased and then decreased (P<0. 05). The protein and mRNA levels of E-cadherin in A549 / DDP cells were lower than those in A549 cells, while the protein and mRNA levels of N-cadherin in A549 / DDP cells were higher than those in A549 cells. The mRNA levels of snail, slug, vimentin, FPn, hepcidin and TfR1 in A549 / DDP cells were higher than those in A549 cells (P<0. 05). The mRNA level of DMT1 in A549 / DDP cells was lower than that in A549 cells (P>0. 05). The cell migration rate was significantly faster in A549 / DDP cells than in A549 cells (P<0. 05). The expression of FtMt in A549 / DDP cells was significantly higher than that in A549 cells (P< 0. 05). Conclusions Cisplatin resistance of lung cancer cells is related to the EMT process, and FtMt might play an important role in cisplatin resistance of lung cancer cells.

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赵峻秀,霍明洋,陈振宇,白雪松,马洪波,郗艳丽.线粒体铁蛋白调控上皮-间质转化对非小细胞肺癌细胞顺铂耐药的影响[J].中国比较医学杂志,2023,33(1):42~50.

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  • 收稿日期:2022-05-04
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  • 在线发布日期: 2023-03-16
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