Inhibitory effect and mechanism of metformin on diffuse large B-cell lymphoma cells

doi:10.11958/20220684

Abstract

Abstract:

Objective To investigate the inhibitory effect and related mechanism of metformin on human diffuse large B-cell lymphoma (DLBCL) U2932 cells. Methods U2932 cells cultured in vitro were treated with different concentrations of metformin, and cells were divided into the control group (0 mmol/L) and the metformin experimental groups (5, 10, 20 and 40 mmol/L). Cells were cultured for 24 h, 48 h and 72 h, respectively. Changes of cell proliferation activity were detected by CCK-8 assay. Cell cycle distribution and apoptosis were detected by flow cytometry. Expression levels of AMPK, CyclinD1, autophagy and apoptosis pathway-related proteins were detected by Western blot assay. Results The proliferation activity of U2932 cells treated with metformin was significantly lower than that of the control group (P<0.05). Compared with the control group, the percentage of cells in G0/G1 phase treated with 5, 10, 20 mmol/L metformin and the apoptosis rate of U2932 cells treated with 20 mmol/L metformin for 24 h, 48 h and 72 h were significantly increased (P<0.05). Western blot assay showed that 5, 10, 20 mmol/L metformin could up-regulate the expression of p-AMPKα, P53, Bax, Cleaved Caspase-3, LC3B proteins, but Beclin1 only partially increased, and down-regulate the expression levels of Bcl-2, p-AKT, CyclinD1 proteins in U2932 cells (P<0.05). Conclusion Metformin can activate AMPK, apoptosis and autophagy pathways, inhibit the proliferation of U2932 cells, and promote their apoptosis.

Key words: metformin, lymphoma, large B-cell, diffuse, AMP-activated protein kinases, apoptosis, cell proliferation, autophagy, signaling pathway

CLC Number:

R733.41

Figures/Tables 8

References 17

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组别	24 h	48 h	72 h	F
对照组	123.77±2.40	204.57±16.26	253.37±7.54	117.969^**
5 mmol/L组	96.63±2.40^a	81.67±1.65^a	61.33±5.15^a	80.585^**
10 mmol/L组	86.87±1.76^ab	75.10±4.78^a	51.37±1.67^ab	102.305^**
20 mmol/L组	69.33±8.00^abc	50.23±1.19^abc	38.97±3.40^abc	27.543^**
40 mmol/L组	61.67±0.97^abcd	39.83±1.10^abc	22.77±0.85^abcd	1 188.112^**
F	112.884^**	224.552^**	1 372.132^**

组别	24 h	48 h	72 h	F
对照组	123.77±2.40	204.57±16.26	253.37±7.54	117.969^**
5 mmol/L组	96.63±2.40^a	81.67±1.65^a	61.33±5.15^a	80.585^**
10 mmol/L组	86.87±1.76^ab	75.10±4.78^a	51.37±1.67^ab	102.305^**
20 mmol/L组	69.33±8.00^abc	50.23±1.19^abc	38.97±3.40^abc	27.543^**
40 mmol/L组	61.67±0.97^abcd	39.83±1.10^abc	22.77±0.85^abcd	1 188.112^**
F	112.884^**	224.552^**	1 372.132^**

组别	G0/G1期	S期	G2/M期
对照组	37.11±1.57	30.65±1.56	30.46±1.46
5 mmol/L组	47.24±1.85^a	27.68±0.77^a	24.63±0.78^a
10 mmol/L组	57.92±1.49^ab	24.42±0.92^ab	17.54±0.81^ab
20 mmol/L组	64.11±9.62^ab	17.97±1.02^abc	12.67±0.79^abc
F	16.906^**	72.239^**	189.181^**

组别	G0/G1期	S期	G2/M期
对照组	37.11±1.57	30.65±1.56	30.46±1.46
5 mmol/L组	47.24±1.85^a	27.68±0.77^a	24.63±0.78^a
10 mmol/L组	57.92±1.49^ab	24.42±0.92^ab	17.54±0.81^ab
20 mmol/L组	64.11±9.62^ab	17.97±1.02^abc	12.67±0.79^abc
F	16.906^**	72.239^**	189.181^**

组别	p-AMPKα	p-AKT	CyclinD1	P53
对照组	1.00±0.00	1.00±0.00	1.00±0.00	1.00±0.00
5 mmol/L组	1.06±0.01^a	0.61±0.01^a	0.18±0.00^a	3.36±0.52^a
10 mmol/L组	1.33±0.01^ab	0.51±0.01^ab	0.17±0.00^ab	8.91±0.35^ab
20 mmol/L组	1.60±0.01^abc	0.21±0.01^abc	0.16±0.00^abc	9.29±0.16^ab
F	5 145.734^**	8 145.469^**	175 735.307^**	484.500^**