中国临床药理学杂志

尼达尼布通过SHP-1介导STAT3信号通路诱导肝癌细胞凋亡及自噬研究

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刘振华 ¹ , 胡永强 ² , 廖斐 ¹ , 赵健 ¹

中国临床药理学杂志 | 临床与基础桥接研究 2025,41(19): 2774-2780

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中国临床药理学杂志 |临床与基础桥接研究 2025 , 41 (19) : 2774 -2780

尼达尼布通过SHP-1介导STAT3信号通路诱导肝癌细胞凋亡及自噬研究

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刘振华¹, 胡永强², 廖斐¹, 赵健¹

作者信息

^1.濮阳医学高等专科学校临床医学学院河南濮阳 457000

^2.濮阳市人民医院肿瘤放疗科河南濮阳 457000

通讯作者:

胡永强，主任医师，硕士研究生导师 MP: 13629865278 E-mail: Ppx2583@163.com

作者简介:

刘振华（1987-），男，本科，讲师，主要负责临床医学方向的工作和研究

Study of nintedanib induces apoptosis and autophagy in hepatocellular carcinoma cells by mediating the STAT 3 signaling pathway through SHP-1

Zhen-hua LIU¹, Yong-qiang HU², Fei LIAO¹, Jian ZHAO¹

Affiliations

^1.School of Clinical Medicine, Puyang Medical College, Puyang 457000, Henan Province, China

^2.Oncology Radiotherapy Department, Puyang City People’s Hospital, Puyang 457000, Henan Province, China

出版时间: 2025-10-17 doi: 10.13699/j.cnki.1001-6821.2025.19.013

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摘要

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目的

探究尼达尼布对肝癌细胞凋亡及自噬的影响，并探究非受体蛋白酪氨酸磷酸酶1（SHP-1）及信号转导和转录激活因子3（STAT3）在其中的作用。

方法

将Hep3B细胞分为Hep3B组（正常培养）、nintedanib组（10 μmol·L^-1尼达尼布处理48 h）、nintedanib+si-NC组（基于nintedanib组，先转染si-NC后再进行尼达尼布处理）、nintedanib+si-SHP-1组（基于nintedanib组，先转染si-SHP-1后再进行尼达尼布处理）。用逆转录实时荧光定量聚合酶链反应法检测SHP-1信使核糖核酸（mRNA）水平；用蛋白质印迹法检测细胞SHP-1、B细胞淋巴瘤-2（Bcl-2）、凋亡抑制因子1（IAP-1）、凋亡相关存活蛋白（Survivin）、苄氯素1（Beclin-1）、磷酸化Beclin-1（p-Beclin-1）、STAT3、磷酸化STAT3（p-STAT3）、Janus激酶2（JAK2）和磷酸化JAK2（p-JAK2）水平；用噻唑蓝实验法检测不同浓度尼达尼布处理后人肝癌细胞Hep3B的细胞活力；用免疫荧光法检测细胞中SHP-1和微管相关蛋白轻链3（LC3）相对表达水平；用原位末端转移酶标记技术法检测各组Hep3B细胞的凋亡情况。

结果

THLE-2、PLC5、HuH7、Hep3B和SK-Hep1各组细胞中SHP-1 mRNA的相对表达水平分别为1.00±0.16、0.45±0.08、0.58±0.09、0.29±0.04和0.34±0.06，SHP-1蛋白的相对表达水平分别为1.00±0.13、0.71±0.09、0.83±0.12、0.56±0.07和0.79±0.08，PLC5、HuH7、Hep3B和SK-Hep1组与THLE-2组相比，SHP-1 mRNA及蛋白相对表达水平在统计学上差异均有统计学意义（均P<0.05）。0、1、5、10、15和20 μmol·L^-1尼达尼布处理后，人肝癌细胞Hep3B的细胞活力分别为（100.00±2.39）%、（92.17±16.85）%、（84.63±15.21）%、（59.02±10.74）%、（36.98±7.53）%和（29.34±5.67）%，除1 μmol·L^-1组外，其余各浓度尼达尼布处理组与0 μmol·L^-1组相比，Hep3B细胞活力在统计学上差异均有统计学意义（均P<0.05）。Hep3B组、nintedanib组、nintedanib+si-NC组和nintedanib+si-SHP-1组细胞中SHP-1 mRNA相对表达水平分别为1.00±0.19、3.06±0.57、2.84±0.52和1.58±0.26，SHP-1蛋白相对表达水平分别为1.00±0.16、3.07±0.54、3.48±0.63和1.32±0.19，细胞的凋亡率分别为（19.34±3.16）%、（62.85±10.93）%、（67.49±11.78）%和（24.72±4.29）%，Bcl-2相对表达水平分别为1.00±0.12、0.56±0.07、0.63±0.08和0.84±0.10，IAP-1相对表达水平分别为1.00±0.15、0.38±0.06、0.44±0.07和0.71±0.12，Survivin相对表达水平分别为1.00±0.14、0.62±0.09、0.56±0.08和0.83±0.11，p-Beclin-1/Beclin-1的水平分别为1.00±0.18、2.37±0.41、2.25±0.39和1.48±0.26，LC3水平分别为1.00±0.21、3.74±0.69、3.15±0.60和1.52±0.28，p-STAT3/STAT3水平分别为1.00±0.11、0.43±0.05、0.49±0.08和0.88±0.10，p-JAK2/JAK2水平分别为1.00±0.15、0.54±0.07、0.48±0.07和0.79±0.13，nintedanib组与Hep3B组相比、nintedanib+si-SHP-1组与nintedanib+si-NC组相比，上述指标在统计学上差异均有统计学意义（均P<0.05）。

结论

尼达尼布可以诱导肝癌细胞凋亡及自噬，这可能是通过SHP-1介导STAT3信号通路实现的。

关键词

尼达尼布 / 肝癌 / 凋亡 / 自噬 / 非受体蛋白酪氨酸磷酸酶1

Abstract

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Objective

To investigate the effects of nintedanib on apoptosis and autophagy of hepatocellular carcinoma cells, and to explore the role of non-receptor type protein tyrosine phosphatase 1(SHP-1) and signal transducer and activator of transcription3 (STAT3).

Methods

Hep3B cells were divided into Hep3B group (normal culture), nintedanib group (treated with 10 μmol·L^-1 nintedanib for 48 h), nintedanib+si-NC group (based on the nintedanib group, transfected with si-NC before nintedanib treatment), and nintedanib+si-SHP-1 group (based on the nintedanib group, transfected with si-SHP-1 before nintedanib treatment). Reverse transcription real-time fluorescence quantitative polymerase chain reaction was used to detect SHP-1 messenger RNA (mRNA) levels. Western blot was used to detect the protein levels of SHP-1, B-cell lymphoma-2 (Bcl-2), inhibitor of apoptosis protein-1 (IAP-1), Survivin, Beclin-1, phosphorylated Beclin-1 (p-Beclin-1), STAT3, phosphorylated STAT3 (p-STAT3), Janus kinase 2 (JAK2) and phosphorylated JAK2 (p-JAK2) in cells. Methyl thiazolyl tetrazolium assay was used to detect the cell viability of human liver cancer cells Hep3B after treatment with different concentrations of nintedanib. Immunofluorescence was used to detect the relative expression levels of SHP-1 and microtubule-associated protein 1 light chain 3 (LC3) in cells. TdT mediated dUTP nick end labeling assay was used to detect the apoptosis of Hep3B cells in each group.

Results

The relative expression levels of SHP-1 mRNA in THLE-2, PLC5, HuH7, Hep3B and SK-Hep1 cells were 1.00±0.16, 0.45±0.08, 0.58±0.09, 0.29±0.04 and 0.34±0.06, respectively; the relative expression levels of SHP-1 protein were 1.00±0.13, 0.71±0.09, 0.83±0.12, 0.56±0.07 and 0.79±0.08, respectively. Compared with the THLE-2 group, the differences in the relative expression levels of SHP-1 mRNA and protein in PLC5, HuH7, Hep3B, and SK-Hep1 groups were statistically significant (all P<0.05). The cell viability of human liver cancer cells Hep3B treated with 0, 1, 5, 10, 15 and 20 μmol·L^-1 nintedanib were (100.00±2.39)%, (92.17±16.85)%, (84.63±15.21)%, (59.02±10.74)%, (36.98±7.53)% and (29.34±5.67)%, respectively. Except for 1 μmol·L^-1 group, the differences in Hep3B cell viability between other nintedanib treatment groups and 0 μmol·L^-1 group were statistically significant (all P<0.05). The relative expression levels of SHP-1 mRNA in Hep3B group, nintedanib group, nintedanib+si-NC group, and nintedanib+si-SHP-1 group cells were 1.00±0.19, 3.06±0.57, 2.84±0.52 and 1.58±0.26, respectively; the relative expression levels of SHP-1 protein were 1.00±0.16, 3.07±0.54, 3.48±0.63 and 1.32±0.19, respectively; the apoptosis rates were (19.34±3.16)%, (62.85±10.93)%, (67.49±11.78)% and (24.72±4.29)%, respectively; the relative expression levels of Bcl-2 were 1.00±0.12, 0.56±0.07, 0.63±0.08 and 0.84±0.10, respectively; the relative expression levels of IAP-1 were 1.00±0.15, 0.38±0.06, 0.44±0.07 and 0.71±0.12, respectively; the relative expression levels of Survivin were 1.00±0.14, 0.62±0.09, 0.56±0.08 and 0.83±0.11, respectively; the levels of p-Beclin-1/Beclin-1 were 1.00±0.18, 2.37±0.41, 2.25±0.39 and 1.48±0.26, respectively; the levels of LC3 were 1.00±0.21, 3.74±0.69, 3.15±0.60 and 1.52±0.28, respectively; the levels of p-STAT3/STAT3 were 1.00±0.11, 0.43±0.05, 0.49±0.08 and 0.88±0.10, respectively; the levels of p-JAK2/JAK2 were 1.00±0.15, 0.54±0.07, 0.48±0.07 and 0.79±0.13, respectively. There were statistically significant differences in the above indicators between nintedanib group and Hep3B group, and between nintedanib+si-shp-1 group and nintedanib+Si NC group (all P<0.05).

Conclusion

Nintedanib can induce apoptosis and autophagy of hepatocellular carcinoma cells, which may be realized through SHP-1 mediated STAT3 signaling pathway.

Key words

nintedanib / hepatocellular carcinoma / apoptosis / autophagy / non-receptor type protein tyrosine phosphatase 1

引用本文

刘振华, 胡永强, 廖斐, 赵健. 尼达尼布通过SHP-1介导STAT3信号通路诱导肝癌细胞凋亡及自噬研究. 中国临床药理学杂志, 2025 , 41 (19) : 2774 -2780 . DOI: 10.13699/j.cnki.1001-6821.2025.19.013

Zhen-hua LIU, Yong-qiang HU, Fei LIAO, Jian ZHAO. Study of nintedanib induces apoptosis and autophagy in hepatocellular carcinoma cells by mediating the STAT 3 signaling pathway through SHP-1[J]. Chinese Journal of Clinical Pharmacology, 2025 , 41 (19) : 2774 -2780 . DOI: 10.13699/j.cnki.1001-6821.2025.19.013

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doi: 10.13699/j.cnki.1001-6821.2025.19.013

接收时间：2025-05-12
首发时间：2026-08-05
出版时间：2025-10-17

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收稿日期：2025-05-12

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作者信息

^1.濮阳医学高等专科学校临床医学学院河南濮阳 457000

^2.濮阳市人民医院肿瘤放疗科河南濮阳 457000

通讯作者:

胡永强，主任医师，硕士研究生导师 MP: 13629865278 E-mail: Ppx2583@163.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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