The effect of dexmedetomidine on the biological behavior of gallbladder cancer cells by regulating the CCL2-CCR2 pathway

doi:10.11958/20252587

Abstract

Abstract:

Objective To explore the effect of dexmedetomidine on the proliferation, invasion and cell cycle of gallbladder cancer cells by regulating the C-C chemokine ligand 2 (CCL2) - C-C chemokine receptor 2 (CCR2) pathway. Methods GBC-SD cells were devided into the control group, the low concentration dexmedetomidine group (2 μmol/L), the high concentration dexmedetomidine group (4 μmol/L) and the high concentration dexmedetomidine+CCL2 group (4 μmol/L dexmedetomidine and 10 μg/L CCL2 protein). The clone formation experiment and Edu experiment were performed to measure cell proliferation. Transwell experiment was performed to measure cell invasion and migration. Flow cytometry was performed to measure cell cycle and apoptosis. Western blot assay was used to measure the proliferating cell nuclear antigen (PCNA), Cyclin D1, matrix metalloproteinase (MMP)-2, MMP-9, Bcl-2 associated X protein (Bax), cysteine-dependent aspartate-specific protease-3 (Caspase-3), CCL2 and CCR2 proteins. The nude mouse transplant tumor experiment was used to determine the growth of gallbladder cancer transplant tumors. Results After treatment with low and high concentrations of dexmedetomidine, the number of cell clone formed, the positive rate of Edu, the numbers of invasions and migrations, the expression levels of PCNA, CyclinD1, MMP-2, MMP-9, CCL2 and CCR2 proteins, the proportions of G2/M and S phase cells decreased, the proportion of G0/G1 phase cells, apoptosis rate and expression levels of Bax and Caspase-3 proteins increased, and the effect of high-concentration dexmedetomidine was more significant (P<0.05). The inhibitory effects of dexmedetomidine on the proliferation, invasion, migration and cell cycle of gallbladder cancer cells, as well as its promoting effect on cell apoptosis could be reversed by CCL2 protein (P<0.05). In vivo experiments showed that dexmedetomidine could reduce tumor mass, tumor volume of nude mice and expression levels of CCL2 and CCR2 proteins (P<0.05). Conclusion Dexmedetomidine inhibits the proliferation and invasion of gallbladder cancer cells, and blocks the cell cycle in the G0/G1 phase by suppressing the CCL2-CCR2 pathway.

Key words: dexmedetomidine, gallbladder neoplasms, cell line, tumor, chemokine CCL2, receptors, CCR2, cell cycle, cell proliferation, cell movement

CLC Number:

R735.8

Figures/Tables 16

References 19

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组别	克隆形成数/个	Edu阳性细胞率/%
对照组	192.63±17.14	45.18±4.93
右美托咪定低浓度组	141.78±13.52^a	31.84±3.64^a
右美托咪定高浓度组	84.25±9.81^ab	17.76±2.06^ab
右美托咪定高浓度+CCL2组	170.04±15.49^c	39.52±4.38^c
F	64.745^＊＊	55.510^＊＊
组别	PCNA	CyclinD1
对照组	1.31±0.14	1.02±0.11
右美托咪定低浓度组	0.96±0.11^a	0.64±0.07^a
右美托咪定高浓度组	0.43±0.05^ab	0.31±0.03^ab
右美托咪定高浓度+CCL2组	1.18±0.12^c	0.87±0.09^c
F	74.305^＊＊	88.185^＊＊

组别	克隆形成数/个	Edu阳性细胞率/%
对照组	192.63±17.14	45.18±4.93
右美托咪定低浓度组	141.78±13.52^a	31.84±3.64^a
右美托咪定高浓度组	84.25±9.81^ab	17.76±2.06^ab
右美托咪定高浓度+CCL2组	170.04±15.49^c	39.52±4.38^c
F	64.745^＊＊	55.510^＊＊
组别	PCNA	CyclinD1
对照组	1.31±0.14	1.02±0.11
右美托咪定低浓度组	0.96±0.11^a	0.64±0.07^a
右美托咪定高浓度组	0.43±0.05^ab	0.31±0.03^ab
右美托咪定高浓度+CCL2组	1.18±0.12^c	0.87±0.09^c
F	74.305^＊＊	88.185^＊＊

组别	侵袭数/个	迁移数/个
对照组	126.83±15.14	142.32±16.38
右美托咪定低浓度组	71.45±8.57^a	87.17±9.84^a
右美托咪定高浓度组	32.72±4.13^ab	41.45±4.66^ab
右美托咪定高浓度+CCL2组	110.24±12.36^c	123.06±14.24^c
F	89.792^＊＊	80.139^＊＊
组别	MMP-2	MMP-9
对照组	1.36±0.13	1.25±0.13
右美托咪定低浓度组	0.91±0.10^a	0.78±0.08^a
右美托咪定高浓度组	0.48±0.05^ab	0.33±0.04^ab
右美托咪定高浓度+CCL2组	1.21±0.13^c	1.04±0.11^c
F	78.065^＊＊	101.924^＊＊

组别	侵袭数/个	迁移数/个
对照组	126.83±15.14	142.32±16.38
右美托咪定低浓度组	71.45±8.57^a	87.17±9.84^a
右美托咪定高浓度组	32.72±4.13^ab	41.45±4.66^ab
右美托咪定高浓度+CCL2组	110.24±12.36^c	123.06±14.24^c
F	89.792^＊＊	80.139^＊＊
组别	MMP-2	MMP-9
对照组	1.36±0.13	1.25±0.13
右美托咪定低浓度组	0.91±0.10^a	0.78±0.08^a
右美托咪定高浓度组	0.48±0.05^ab	0.33±0.04^ab
右美托咪定高浓度+CCL2组	1.21±0.13^c	1.04±0.11^c
F	78.065^＊＊	101.924^＊＊

组别	G0/G1期	G2/M期	S期
对照组	26.18±2.93	24.74±2.81	49.08±5.02
右美托咪定低浓度组	41.42±4.71^a	16.85±1.84^a	41.73±4.32^a
右美托咪定高浓度组	54.83±5.90^ab	11.26±1.42^ab	33.91±3.27^ab
右美托咪定高浓度+CCL2组	33.25±3.67^c	21.57±2.58^c	45.18±5.01^c
F	45.930^＊＊	41.478^＊＊	12.540^＊＊