6-姜辣素对人多发性骨髓瘤细胞的抑制作用及分子机制

doi:10.3969/j.issn.1006-5725.2024.23.003

摘要/Abstract

摘要：

目的研究6-姜辣素对人多发性骨髓瘤细胞的抑制作用及分子机制。方法体外培养人多发性骨髓瘤RPMI 8226、ARH77细胞，加入不同浓度（50、100、200、300、400 μmol/L）的6-姜辣素处理RPMI 8226、ARH77 细胞，CCK-8法测定细胞增殖抑制率；流式细胞术检测细胞凋亡和周期；qRT-PCR和Western blotting法检测基因和蛋白表达水平。结果 6-姜辣素以剂量和时间依赖的方式抑制RPMI 8226、ARH77细胞的增殖，并诱导其凋亡，差异有统计学意义（P < 0.05）；进一步机制研究发现6-姜辣素处理RPMI 8226细胞后，将细胞阻滞在G₀/G₁期，显著增加Bax mRNA水平，降低Bcl-2 mRNA和c-Myc mRNA的水平（P < 0.05）；同时显著增加Bax、Cleaved-PARP、Cleaved-caspase3、P53、p-AKT蛋白的表达，而降低Bcl-2蛋白的表达（P < 0.05）。结论 6-姜辣素可抑制MM细胞增殖及诱导其凋亡，并阻滞细胞周期在G₀/G₁期，其机制可能与抑制AKT信号通路并通过影响 Bcl-2家族蛋白表达以及c-Myc表达受抑等密切有关。

关键词: 多发性骨髓瘤, 6-姜辣素, 细胞增殖, 细胞凋亡, 信号通路

Abstract:

Objective To investigate the inhibitory effect and elucidate the molecular mechanism of 6?gingerol on human multiple myeloma cells. Methods The human multiple myeloma cell lines RPMI 8226 and ARH77 were cultured in vitro， followed by treatment with varying concentrations （50， 100， 200， 300， 400 μmol/L） of 6?gingerol. The inhibitory effect on cell proliferation was assessed using the CCK?8 assay. Flow cytometry was employed to evaluate cell apoptosis and cycle distribution. Additionally， qRT?PCR and Western blotting techniques were utilized to analyze gene and protein expression levels. Results The proliferation of RPMI 8226 and ARH77 cells was dose? and time?dependently inhibited by 6?gingerol， leading to the induction of apoptosis with statistically significant differences （P < 0.05）. Further mechanistic investigations revealed that treatment with 6?gingerol arrested RPMI 8226 cells in the G0/G1 phase， resulting in a significant increase in Bax levels and a decrease in Bcl?2 mRNA and c?Myc mRNA levels （P < 0.05）. Additionally， it significantly upregulated the expression of Bax， Cleaved?PARP， Cleaved?caspase3， P53， and p?AKT proteins while down regulating the expression of Bcl?2 protein （P < 0.05）. Conclusions The compound 6?Gingerol exhibits inhibitory effects on the proliferation and induction of apoptosis in MM cells， as well as cell cycle arrest at the G0/G1 phase. Its mechanism of action is likely associated with the suppression of the AKT signaling pathway， downregulation of Bcl?2 family protein expression， and inhibition of c?Myc expression.

Key words: multiple myeloma, 6-gingerol, cell proliferation, cell apoptosis, signal pathway

中图分类号:

R559

孔春芳,李安娜,柯波,丁伟荣,刘婷婷,符环,张婷婷,金成豪,吴美. 6-姜辣素对人多发性骨髓瘤细胞的抑制作用及分子机制[J]. 实用医学杂志, 2024, 40(23): 3291-3297.

Chunfang KONG,Anna LI,Bo KE,Weirong DING,Tingting LIU,Huan FU,Tingting ZHANG,Chenghao JIN,Mei. WU. The inhibitory effect and molecular mechanism of 6-gingerol on human multiple myeloma cells[J]. The Journal of Practical Medicine, 2024, 40(23): 3291-3297.

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时间	DMSO	50 μmol/L	100 μmol/L	200 μmol/L	300 μmol/L	400 μmol/L	F值	P值
24 h	0.00 ± 0.00	3.05 ± 0.87^*	5.18 ± 0.42^**	9.88 ± 0.49^**	16.19 ± 0.72^**	27.73 ± 0.72^**	951.30	< 0.001
48 h	0.00 ± 0.00	3.88 ± 0.75^**	8.15 ± 0.28^**	17.72 ± 0.68^**	27.68 ± 0.63^**	44.94 ± 2.71^**	587.63	< 0.001
72 h	0.00 ± 0.00	8.31 ± 1.16^**	13.29 ± 1.12^**	25.54 ± 1.31^**	36.32 ± 1.15^**	60.48 ± 1.01^**	1298.38	< 0.001

时间	DMSO	50 μmol/L	100 μmol/L	200 μmol/L	300 μmol/L	400 μmol/L	F值	P值
24 h	0.00 ± 0.00	3.32 ± 0.62^*	7.77 ± 0.51^***	14.43 ± 1.21^**	27.36 ± 0.62^**	35.91 ± 0.60^**	1 463.74	< 0.001
48 h	0.00 ± 0.00	4.93 ± 0.42^*	10.53 ± 0.41^***	21.45 ± 1.62^***	39.14 ± 0.69^***	61.42 ± 1.85^***	1 438.98	< 0.001
72 h	0.00 ± 0.00	9.12 ± 0.78^***	17.58 ± 0.88^***	31.05 ± 1.29^***	48.53 ± 1.42^***	76.19 ± 0.93^***	1 409.23	< 0.001

项目	DMSO	50 μmol/L	100 μmol/L	200 μmol/L	300 μmol/L	400 μmol/L	F值	P值
ARH77	2.60 ± 0.41	7.86 ± 1.47^**	10.20 ± 1.00^**	12.71 ± 1.17^***	19.47 ± 1.79^***	39.23 ± 3.12^***	171.48	< 0.001
RPMI?8226	3.66 ± 0.44	7.92 ± 0.67^*	11.09 ± 0.84^**	16.99 ± 1.20^***	31.69 ± 1.22^***	56.56 ± 0.85^***	1420.24	< 0.001

细胞周期	DMSO	300 μmol/L	400 μmol/L	F值	P值
G₁期	16.90 ± 0.86	30.95 ± 1.06^***	40.31 ± 1.90^****	228.41	< 0.001
S期	64.11 ± 0.60	61.44 ± 5.88	48.82 ± 0.46^***	17.08	0.003
G₂期	18.96 ± 0.96	7.61 ± 4.81^*	10.88 ± 2.35^**	10.78	0.011

项目	DMSO	300 μmol/L	400 μmol/L	F值	P值
Bcl-2	1	0.744 ± 0.055^*	0.108 ± 0.012^****	591.68	< 0.001
C-myc	1	0.817 ± 0.079^*	0.714 ± 0.079^**	75.20	0.004
Bax	1	1.260 ± 0.101^*	3.427 ± 0.485^***	65.02	< 0.001