绞股蓝皂苷LI通过代谢脂肪酸通路下调肉碱棕榈酰转移酶1B抑制结肠癌生长

doi:10.3969/j.issn.1006-5725.2025.02.002

Abstract

Abstract:

Objective To elucidate the mechanism through which Gypenoside L inhibits the growth of colon cancer by modulating carnitine palmitoyltransferase 1B （CPT1B）， a pivotal enzyme in the fatty acid metabolism pathway. Methods Through in vitro experiments， various concentrations of Gypenoside LI LI were applied to intervene in colon cancer RKO and SW620 cells. The effects of Gypenoside LI on these cells were comprehensively evaluated using the CCK?8 assay， wound healing assay， colony formation assay， and live?dead cell staining， focusing on its impact on cell proliferation， migration， and apoptosis. Additionally， a human colon cancer tissue microarray （TMA） was utilized in conjunction with multiplex fluorescence immunohistochemistry to analyze the expression of CPT1B in colon cancer and adjacent tissues. SW620 cells were transfected with siRNA， and the mRNA and protein expression levels of CPT1B post?transfection were assessed using quantitative real?time PCR （qPCR） and Western blotting. Furthermore， an in vivo nude mouse colon cancer model was established to investigate the inhibitory effect of Gypenoside LI LI on colon cancer growth. Results In vitro experiments demonstrated that Gypenoside LI LI effectively inhibited the proliferation and migration of RKO and SW620 cells in a concentration? and time?dependent manner. Additionally， multiple fluorescence immunohistochemistry analyses revealed that the expression level of CPT1B in colon cancer tissues was significantly higher than that in adjacent non?tumor tissues. Gypenoside LI LI promoted ROS accumulation by inhibiting CPT1B expression. In vivo experiments further confirmed that Gypenoside LI LI could inhibit tumor formation in nude mice and reduce CPT1B expression. Conclusions This study elucidates the mechanism by which Gypenoside LI inhibits the growth of colon cancer cells. Specifically， it downregulates CPT1B， leading to increased accumulation of reactive oxygen species （ROS）， disruption of fatty acid oxidation metabolism， and ultimately inducing apoptosis in colon cancer cells. These findings offer valuable insights into colon cancer treatment， suggesting new therapeutic strategies and potential drug targets.

Key words: Gypenoside LI, carnitine Palmitoyltransferase 1b （CPT1B）, colon cancer, fatty acid metabolism, drug target

CLC Number:

R285.6

Wenyu ZHU,Hongwei ZHANG,Decai TANG,Fangyuan CHEN,Hua JIANG,Haiyan MIN,Jie. DING. Gypenosides LI down⁃regulates CPT1B through the pathway of lipid metabolism to inhibit the growth of colon cancer[J]. The Journal of Practical Medicine, 2025, 41(2): 162-169.

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References 28

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名称	序列
名称	Sense（5′?3′）（包括overhangs）	Antisense（5′?3′）（包括overhangs）
si?homo?CPT1B?1	GUUCUCAAGCUCAGAGACGAA	CGUCUCUGAGCUUGAGAACUU
si?homo?CPT1B?2	GUACCUAGGAGUCAGCUCUCC	AGAGCUGACUCCUAGGUACUU
si?homo?CPT1B?3	GGACACAGAUGUGCUACAGCA	CUGUAGCACAUCUGUGUCCUU
si?NC	UUCUCCGAACGUGUCACGUTT	ACGUGACACGUUCGGAGAATT

Gypenoside LI浓度	RKO细胞活力（48 h）	RKO细胞活力（72 h）
对照组	100.00 ± 1	100.00 ± 0.57
0.1 mg/mL	97.00 ± 24	114.67 ± 34.02
0.25 mg/mL	90.00 ± 11	102.67 ± 25.50
0.5 mg/mL	84.33 ± 5.5	77.00 ± 15.00
1 mg/mL	82.00 ± 6	66.67 ± 24.50^#
2 mg/mL	30.33 ± 0.57^?#&∧$	11.67 ± 0.58^?#&∧$
F值	15.42	13.39
P值	< 0.001	0.001

Gypenoside LI浓度	SW620细胞活力（48 h）	SW620细胞活力（72 h）
对照组	100.00 ± 2.70	100.00 ± 2.51
0.1 mg/mL	90.17 ± 3.20^?	98.57 ± 8.55
0.25 mg/mL	72.83 ± 2.55^?#	98.33 ± 5.75
0.5 mg/mL	76.20 ± 2.35^?#	92.17 ± 5.55
1 mg/mL	68.87 ± 5.82^?#	68.30 ± 2.82^?#&∧
2 mg/mL	16.37 ± 3.25^?#&∧$	30.17 ± 1.50^?#&∧$
F值	203.7	89.51
P值	< 0.001	< 0.001

Gypenoside LI浓度	SW620细胞迁移距离	RKO细胞迁移距离
对照组	189.62 ± 13.25	62.97 ± 4.28
0.05 mg/mL	100.54 ± 6.29^?	99.00 ± 8.49^?
0.1 mg/mL	113.78 ± 7.37^?	75.39 ± 3.69
0.2 mg/mL	9.00 ± 5.65^?#&	74.50 ± 6.36
F值	145.4	12.66
P值	< 0.001	< 0.05

Gypenoside LI浓度	RKO细胞克隆数目	SW620细胞克隆数目
0.05 mg/mL	377.50 ± 103.94	1750.00 ± 127.27
0.1 mg/mL	505.00 ± 107.48	1660.00 ± 84.85
0.2 mg/mL	47.15 ± 14.35^?#	6.76 ± 0.33^?#
F值	14.85	247.00
P值	< 0.05	< 0.001

Gypenosides LI down⁃regulates CPT1B through the pathway of lipid metabolism to inhibit the growth of colon cancer

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Gypenoside LI浓度	相对AM/PI比值（RKO细胞）	相对AM/PI比值（SW620细胞）
对照	1.03 ± 0.15	1.00 ± 0.10
0.1 mg/mL	0.44 ± 0.06^?	0.42 ± 0.072^?
0.2 mg/mL	0.004 ± 0.001^?#	0.034 ± 0.058^?#
F值	87.45	114.9
P值	< 0.001	< 0.001

Gypenoside LI浓度	平均荧光强度
对照组	54.67 ± 4.51
0.05 mg/mL	104.00 ± 7.94
0.1 mg/mL	385.00 ± 37.00^?#
0.2 mg/mL	612.00 ± 29.61^?#&
F值	348.1
P值	< 0.001

分组	平均荧光强度
对照组	44.00 ± 3.00
Gypenoside LI	379.33 ± 16.86^?
si-CPT1B	431.00 ± 30.45^?
Gypenoside LI+si-CPT1B	550.67 ± 32.87^?#&
F值	245.70
P值	<0.001

Gypenoside LI浓度	CPT1B相对表达量	CPT2相对表达量
PBS	1.00 ± 0.02	1.00 ± 0.02
10 mg/mL	0.83 ± 0.02^?	1.82 ± 0.13^?
20 mg/mL	0.51 ± 0.02^?#	2.10 ± 0.12^?#
40 mg/mL	0.24 ± 0.01^?#&	2.14 ± 0.08^?#
F值	987.40	88.87
P值	< 0.001	< 0.001

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