亚精胺对弥漫大B细胞淋巴瘤细胞株增殖和凋亡的影响

doi:10.3969/j.issn.1006-5725.2024.22.003

Abstract

Abstract:

Objective To investigate the impact of spermidine on proliferation and apoptosis of diffuse large B?cell lymphoma （DLBCL） cell lines. Methods The impact of spermidine on cellular growth was assessed using a CCK?8 assay. Flow cytometry was employed to investigate the effects of spermidine on the proliferation and cell cycle dynamics of DLBCL cell lines， as well as to evaluate its influence on apoptosis in DLBCL cell lines， mouse splenocytes， and peripheral blood mononuclear cells （PBMCs） derived from healthy individuals. Western blot analysis was conducted to examine alterations in protein expression levels associated with apoptosis and the cell cycle following treatment with spermidine. Results The CCK?8 assay revealed a significant inhibitory effect of spermidine on DLBCL cell growth （P < 0.001）. Flow cytometric analysis demonstrated that spermidine had no impact on the proliferation or cell cycle of DLBCL cells， but significantly induced apoptosis （P < 0.001）. Spermidine exhibited a pro?apoptotic effect on mouse splenocytes （P < 0.01）， albeit weaker compared to its effect on DLBCL cells （P < 0.001）， and showed no significant pro?apoptotic effect on PBMCs. Western blot results indicated that spermidine did not influence the expression levels of cell cycle proteins CDK2 and CDK4， but enhanced the activation of Caspase?9 in A20 cells and Caspase?8 in OCI?Ly3 cells. Conclusion Spermidine induces apoptosis and suppresses cell growth in DLBCL cell lines， while exhibiting diminished or absent pro-apoptotic effects on mouse splenocytes and healthy human PBMCs， suggesting its potential as a specific inhibitor for the growth of DLBCL cell lines in vivo.

Key words: spermidine, diffuse large B-cell lymphoma, proliferation, apoptosis

CLC Number:

R733.4

Bing′er WU,Qing LI,Kerong YANG,Jian ZHANG,Yi YU,Lei LEI,Bo. HU. Impact of spermidine on proliferation and apoptosis in diffuse large B⁃cell lymphoma cell lines[J]. The Journal of Practical Medicine, 2024, 40(22): 3130-3137.

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组别	24 h OD值（450 nm）	48 h OD值（450 nm）
Control组	0.254 ± 0.004	0.409 ± 0.012
2.5 μmol/L Spermidine组	0.251 ± 0.008^**	0.290 ± 0.024^##***
5 μmol/L Spermidine组	0.237 ± 0.006^#	0.246 ± 0.004^##*
10 μmol/L Spermidine组	0.233 ± 0.006^##	0.224 ± 0.003^##
F值	17.22	190.0
P值	< 0.000 1	< 0.000 1

组别	24 h OD值（450 nm）	48 h OD值（450 nm）
Control组	0.366 ± 0.015	0.724 ± 0.103
2.5 μmol/L Spermidine组	0.332 ± 0.013^#**	0.593 ± 0.127^**
5 μmol/L Spermidine组	0.306 ± 0.016^###**	0.482 ± 0.044^##*
10 μmol/L Spermidine组	0.248 ± 0.014^###	0.271 ± 0.003^###
F值	53.04	25.74
P值	< 0.000 1	< 0.000 1

组别	A20（48 h）		OCI?Ly3（48 h）
组别	Ki67/%	Ki67（MFI）	Ki67/%	Ki67（MFI）
Control组	92.274 ± 0.646	40 591.600 ± 1 246.350	94.050 ± 0.545	48 532.250 ± 4 864.604
10 μmol/L Spermidine组	92.570 ± 0.130	42 887.800 ± 1 753.880	94.262 ± 0.633	52 863.400 ± 3 002.796
t值	1.004	2.386	1.495	0.144
P值	0.345	0.044	0.613	0.143

组别	A20（48 h）			OCI-Ly3（48 h）
组别	G₀/G₁	S	G₂/M	G₀/G₁	S	G₂/M
Control组	60.378 ± 3.048	34.152 ± 1.581	1.872 ± 1.339	66.234 ± 3.087	22.088 ± 2.066	10.554 ± 0.917
10 μmol/L Spermidine组	57.884 ± 2.234	33.808 ± 1.836	3.504 ± 0.867	62.994 ± 3.504	24.594 ± 2.211	11.284 ± 2.418
t值	1.476	0.318	2.287	1.551	1.851	0.631
P值	0.179	0.759	0.052	0.159	0.101	0.546

组别	A20（48 h）		OCI?Ly3（48 h）
组别	CDK2	CDK4	CDK2	CDK4
Control组	1.242 ± 0.432	1.583 ± 0.324	1.301 ± 0.350	1.409 ± 0.414
10 μmol/L Spermidine组	1.178 ± 0.323	1.381 ± 0.269	1.283 ± 0.341	1.560 ± 0.373
t值	0.206	0.827	0.063	0.468
P值	0.847	0.455	0.953	0.665

Impact of spermidine on proliferation and apoptosis in diffuse large B⁃cell lymphoma cell lines

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组别	A20（48 h）			OCI?Ly3（48 h）
组别	早期凋亡细胞	晚期凋亡细胞	总凋亡细胞	早期凋亡细胞	晚期凋亡细胞	总凋亡细胞
Control组	1.465 ± 0.452	2.673 ± 0.859	4.138 ± 1.298	5.258 ± 0.457	3.430 ± 0.510	8.688 ± 0.350
10 μmol/L Spermidine组	40.595 ± 7.130	25.643 ± 4.750	66.238 ± 11.449	16.245 ± 2.856	8.245 ± 1.434	24.490 ± 1.868
t值	17.20	11.06	16.02	7.597	6.326	16.63
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	A20（48 h）		OCI?Ly3（48 h）
组别	Caspase?8	Caspase?9	Caspase?8	Caspase?9
Control组	1.322 ± 0.225	1.583 ± 0.324	1.557 ± 0.270	1.282 ± 0.536
10 μmol/L Spermidine组	1.222 ± 0.316	2.808 ± 0.448	2.956 ± 0.423	1.357 ± 0.430
t值	0.449	2.887	0.290	0.397
P值	0.677	< 0.05	< 0.01	0.859

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组别	小鼠脾脏细胞			人PBMCs（48 h）
组别	早期凋亡细胞	晚期凋亡细胞	总凋亡细胞	早期凋亡细胞	晚期凋亡细胞	总凋亡细胞
Control组	15.854 ± 0.527	11.640 ± 1.894	27.494 ± 2.276	6.360 ± 0.671	6.534 ± 1.788	12.894 ± 2.437
10 μmol/L Spermidine组	19.112 ± 1.508	13.142 ± 3.331	32.254 ± 4.686	6.840 ± 0.507	5.806 ± 0.941	12.646 ± 1.401
t值	5.248	3.957	9.787	1.276	0.806	0.197
P值	< 0.01	0.406	0.075	0.238	0.444	0.849