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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

27卷

期数:

2021年03期

页码:

211-216

栏目:

基础医学

出版日期:

2021-05-30

文章信息/Info

Title:

Gefitinib inhibits the proliferation of malignant peripheral nerve sheath tumor cells by promoting H3K27 methylation levels

文章编号:

1006-8147(2021)03-0211-06

作者:

孙硕遥¹; 高雅¹; 朱香熹²; 赵洋¹; 赵玉龙¹; 李光明¹; 杨吉龙³; 朱泽¹

1.天津医科大学基础医学院病原生物学系，天津300070；2.遵义医科大学珠海校区临床医学系,珠海519090；3.天津医科大学肿瘤医院骨与软组织肿瘤科，天津300060

Author(s):

SUN Shuo-yao¹; GAO Ya¹; ZHU Xiang-xi²; ZHAO Yang¹; ZHAO Yu-long¹; LI Guang-ming¹; YANG Ji-long³; ZHU Ze¹

1.Department of Pathogen Biology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China；2.Department of Clinical Medicine， Zhuhai Campus of Zunyi Medical University， Zhuhai 519090，China；3.Department of Bone and Soft Tissue Tumor，Cancer Institute and Hospital，Tianjin Medical University，Tianjin 300060，China

关键词:

恶性周围神经鞘瘤; H3K27me3; Kras; EGFR; 吉非替尼

Keywords:

malignant peripheral nerve sheath tumor; H3K27me3; Kras; EGFR; gefitinib

分类号:

R739.43

DOI:

-

文献标志码:

A

摘要:

目的：研究表皮生长因子受体（EGFR）抑制剂吉非替尼（gefitinib）对恶性周围神经鞘瘤（MPNST）细胞增殖、迁移的影响。方法：选择STS26T细胞、ST88-14细胞，分为对照组和给药组（10 μmol/L），其中药物IC50实验、蛋白印迹实验给药组浓度分为5、10、15 μmol/L。CCK-8法检测吉非替尼对STS26T、ST88-14的半数抑制浓度并绘制药物IC50曲线；平板克隆形成实验检测吉非替尼对细胞增殖能力的影响；划痕实验检测吉非替尼对细胞迁移能力的影响；实时荧光定量PCR检测加药后各组细胞Kras、SUZ12、EED、EZH2 mRNA的表达情况；蛋白印迹实验检测加药后Kras、组蛋白H327位赖氨酸三甲基化蛋白（H3K27me3）的表达。结果：STS26T、ST88-14细胞的药物IC50曲线显示半数抑制浓度均为10 μmol/L（t=11.42、16.51，均P＜0.05）；与对照组比较，吉非替尼给药组（10 μmol/L）STS26T、ST88-14细胞增殖能力显著降低（t=5.48，P＜0.05；t=4.89，P＜0.01）、细胞迁移能力显著降低（t=4.94，P＜0.01；t=4.75，P＜0.01）;与对照组比较，吉非替尼给药组（10 μmol/L）STS26T细胞中Kras mRNA表达显著降低（t=4.87，P<0.01），SUZ12、EED、EZH2表达显著升高（t=11.36、15.54、13.19，均P＜0.05）；与对照组比较，吉非替尼给药组（10 μmol/L）ST88-14细胞中Kras mRNA表达显著降低（t=13.75，P<0.05），SUZ12、EED、EZH2表达水平显著升高（t=12.56、7.48，16.33，均P＜0.05）；与对照组比较，吉非替尼给药组（5、10、15 μmol/L）STS26T、ST88-14细胞Kras蛋白表达显著降低（t=13.70、15.21，均P<0.05）；与对照组比较，吉非替尼给药组（5、10、15 μmol/L）STS26T、ST88-14细胞H3K27me3蛋白表达显著增加（t=14.31、12.40，均P<0.05）。结论：EGFR抑制剂吉非替尼通过促进MPNST中PRC2的表达，提高表观遗传学标志物H3K27me3的表达，降低MPNST细胞株增殖、迁移的能力。

Abstract:

Objective: To study the effect of epidermal growth factor receptor（EGFR） inhibitor gefitinib on the proliferation and migration of malignant peripheral nerve sheath tumor（MPNST） cells. Methods: STS26T cells and ST88-14 cells were selected and divided into control group and administration group（10 μmol/L）. The administration groups of drug IC50 experiment and Western blotting experiment were divided into 5，10 and 15 μmol/L. Using CCK-8 method was used to identify the half inhibitory concentration of gefitinib on STS26T and ST88-14， and the drug IC50 curve was drawn. The effect of gefitinib on cell proliferation was detected by colony formation assay. Wound healing test was used to detect the effect of gefitinib on cell migration.Real-time fluorescence quantification PCR was used to detect the expression of Kras，SUZ12，EED，EZH2 mRNA in each group of cells after adding drugs.Western blotting was used to detect the expression of Kras and histone H3K27 lysine trimethylated（H3K27me3）protein after adding drugs. Results: The drug IC50 curve showed that the half inhibitory concentration of STS26T and ST88-14 was both 10 μmol/L（t=11.42，16.51，all P＜0.05）.Compared with the control group， the proliferation and migration ability of STS26T and ST88-14 cells in the gefitinib administration group（10 μmol/L） was significantly reduced（t=5.48，P＜0.05；t=4.89，P＜0.01；t=4.94，P<0.01；t=4.75，P<0.01）. Compared with the control group， the mRNA expression of Kras in the gefitinib administration group（10 μmol/L） of STS26T cells was significantly reduced（t=4.87，P<0.01）， and the expression of SUZ12， EED， and EZH2 were significantly increased（t=11.36，15.54，13.19， all P<0.05）. Compared with the control group， the mRNA expression of Kras in the gefitinib administration group（10 μmol/L） of ST88-14 cells was significantly reduced （t=13.75，P<0.05）， and the mRNA expression of levels SUZ12，EED，and EZH2 were significantly increased（t=12.56，7.48，16.33，all P<0.05）. Compared with the control group， the expression of Kras protein of STS26T and ST88-14 cells in the gefitinib administration group（5，10，15 μmol/L） was significantly reduced（t=13.70，t=15.21，all P<0.05）.Compared with the control group，the expression of H3K27me3 protein of STS26T and ST88-14 cells in the gefitinib administration group（5，10，15 μmol/L） was significantly increased（t=14.31，12.40，all P<0.05）.Conclusion: EGFR targeting inhibitor gefitinib increases the expression of the epigenetic marker H3K27me3 by promoting the expression of PRC2 in MPNST， and reduces the proliferation，migration of MPNST cell lines.

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备注/Memo

备注/Memo:

基金项目 国家自然科学基金（81672650）
作者简介 孙硕遥(1995-)，女，硕士在读，研究方向：恶性外周神经鞘瘤发病与发展的分子机制；
通信作者：朱泽，E-mail: zhuze@tijmu.edu.

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更新日期/Last Update: 2021-05-30