三七总皂苷对胃癌细胞生物学行为和Keap1-Nrf2信号通路的影响

doi:10.13481/j.1671-587X.20260509

Abstract

Abstract:

Objective To discuss the effects of Panax notoginseng saponins （PNS） on the biological behavior of gastric cancer （GC） cells and to analyze its relationship with the Kelch-like ECH-associated protein 1 （Keap1）-nuclear factor E2-related factor 2 （Nrf2） signaling pathway. Methods GC cell lines AGS and HGC27 cells and human normal gastric mucosal epithelial cells GES-1 were used as the research objects and treated with different concentrations of PNS. MTT method was used to detect the cell activity in various groups. AGS and HGC27 cells were divided into control group （0 μmol·L^-1 PNS group） and 5， 10， and 20 μmol·L^-1 PNS groups. Colony formation assay was used to detect the proliferation abilities of the cells in various groups. Transwell chamber assay was used to detect the numbers of migration cells and invasion cells in various groups. 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescent probe method was used to detect the reactive oxygen species （ROS） levels in the cells in various groups. Western blotting method was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， Bcl-2-associated death promoter （Bad）， cysteinyl aspartate-specific proteinase 3 （Caspase-3）， and cleaved Caspase-3， as well as Keap1 and Nrf2 proteins in the cells in various groups. Co-immunoprecipitation （Co-IP） method combined with Western blotting method was used to detect the interaction between Keap1 and Nrf2 and Nrf2 ubiquitination in the cells in various groups. Results After treatment for 48 h， compared with control group， the activities of AGS and HGC27 cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly decreased （P<0.001）， while there was no statistically significant difference in the activity of GES-1 cells （P>0.05）. Compared with control group， the colony formation numbers， numbers of migration cells， and numbers of invasion cells of AGS and HGC27 cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly decreased （P<0.001）， while the apoptotic rates and the ROS levels in the cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly increased （P<0.05 or P<0.01）. Compared with control group， the expression levels of Bcl-2 and Nrf2 proteins in AGS and HGC27 cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly decreased （P<0.001）， while the expression levels of Bad， Bax， cleaved Caspase-3， and Keap1 proteins were significantly increased （P<0.001）. Compared with control group， the co-precipitation signals of Keap1 and Nrf2 in AGS and HGC27 cells in PNS group were increased， while the ubiquitination signals of Nrf2 in AGS and HGC27 cells were decreased. Conclusion PNS inhibits the proliferation， migration， and invasion of GC cells， promotes cell apoptosis， and enhances intracellular oxidative stress level. Its effect is related to the regulation of the expression of Keap1-Nrf2 signaling pathway-related proteins and the interaction between Keap1 and Nrf2.

Key words: Panax notoginseng saponins, Gastric cancer, Apoptosis, Reactive oxygen species, Oxidative stress, Kelch-like ECH-associated protein 1, Nuclear factor E2-related factor 2

CLC Number:

R285.5

Dongyu XIANG,Guiyu CHEN,Yue XIANG. Effects of Panax notoginseng saponins on biological behaviors of gastric cancer cells and Keap1-Nrf2 signaling pathway[J].Journal of Jilin University(Medicine Edition), 2026, 52(5): 1273-1281.

Figures/Tables 11

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References 19

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Cell line	Group	Cell viability		F	P
Cell line	Group	（t/h） 24	48	F	P
GES-1	Control	99.58±2.73	100.31±2.88	1.083	0.379
	5 μmol·L^-1 PNS	101.84±3.12	95.76±3.04
	10 μmol·L^-1 PNS	107.36±3.78	92.68±2.83
	20 μmol·L^-1 PNS	109.62±4.21	91.92±3.18
AGS	Control	99.82±2.61	100.47±2.79	12.468	<0.001
	5 μmol·L^-1 PNS	97.91±2.94	81.68±3.56^*
	10 μmol·L^-1 PNS	84.63±3.69^*	67.94±3.87^*
	20 μmol·L^-1 PNS	64.77±4.08^*	41.63±3.36^*
HGC27	Control	99.91±2.53	100.38±2.66	14.021	<0.001
	5 μmol·L^-1 PNS	91.76±3.19^*	79.61±3.48^*
	10 μmol·L^-1 PNS	81.58±3.55^*	63.72±3.82^*
	20 μmol·L^-1 PNS	65.87±3.98^*	37.88±3.29^*

Group	AGS			HGC27
Group	Number of colony formation	Number of migration cells	Number of invasion cells	Number of colony formation	Number of migration cells	Number of invasion cells
Control	172.40±10.85	42.30±3.85	43.50±3.96	131.60±9.42	43.10±3.92	44.20±4.01
5 μmol·L^-1 PNS	142.10±9.76^*	34.20±3.01^*	32.60±2.88^*	105.80±8.31^*	34.60±3.08^*	31.80±2.74^*
10 μmol·L^-1 PNS	96.30±8.14^*	20.10±2.14^*	15.30±1.92^*	78.90±6.87^*	19.40±2.05^*	16.10±1.87^*
20 μmol·L^-1 PNS	68.70±7.02^*	12.40±1.63^*	9.80±1.41^*	52.40±5.96^*	13.10±1.58^*	11.40±1.53^*
F	58.926	96.284	121.639	64.381	102.517	118.402
P	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001

Group	AGS					HGC27
Group	Bcl-2	Bad	Bax	Caspase-3	Cleaved Caspase-3	Bcl-2	Bad	Bax	Caspase-3	Cleaved Caspase-3
Control	1.00±0.07	1.00±0.08	1.00±0.07	1.00±0.06	1.00±0.08	1.00±0.06	1.00±0.07	1.00±0.07	1.00±0.06	1.00±0.07
5 μmol·L^-1 PNS	0.55±0.05^*	1.25±0.09^*	1.15±0.08^*	1.05±0.07	1.15±0.09^*	0.80±0.06^*	1.20±0.09^*	1.15±0.08^*	1.03±0.06	1.10±0.08^*
10 μmol·L^-1 PNS	0.48±0.04^*	1.60±0.12^*	1.50±0.11^*	1.02±0.06	1.60±0.12^*	0.75±0.05^*	1.55±0.11^*	1.60±0.12^*	1.00±0.06	1.65±0.12^*
20 μmol·L^-1 PNS	0.42±0.04^*	1.75±0.13^*	1.80±0.14^*	1.08±0.07	2.10±0.15^*	0.70±0.05^*	1.80±0.13^*	1.75±0.13^*	1.05±0.07	1.95±0.14^*
F	132.418	88.264	104.596	1.214	146.372	96.537	91.173	112.483	0.982	139.824
P	<0.001	<0.001	<0.001	0.342	<0.001	<0.001	<0.001	<0.001	0.415	<0.001

Group	AGS		HGC27
Group	Keap1	Nrf2	Keap1	Nrf2
Control	1.00±0.08	1.00±0.07	1.00±0.07	1.00±0.06
5 μmol·L^-1 PNS	2.05±0.16^*	0.90±0.07^*	1.25±0.10^*	0.82±0.06^*
10 μmol·L^-1 PNS	2.65±0.21^*	0.60±0.05^*	1.60±0.12^*	0.60±0.05^*
20 μmol·L^-1 PNS	3.00±0.24^*	0.50±0.04^*	2.10±0.16^*	0.30±0.03^*
F	128.437	96.214	112.583	104.762
P	<0.001	<0.001	<0.001	<0.001

Effects of Panax notoginseng saponins on biological behaviors of gastric cancer cells and Keap1-Nrf2 signaling pathway

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