The effects of daphnetin regulating the Src/FAK pathway on the proliferation, migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells

doi:10.11958/20260416

Abstract

Abstract:

Objective To investigate the effects of daphnetin regulating the steroid receptor coactivator (Src)/focal adhesion kinase (FAK) pathway on the proliferation, migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells. Methods Human nasopharyngeal carcinoma cells 5-8F were assigned into the control group, the L-daphnetin (4 mg/L) group, the M-daphnetin (8 mg/L) group, the H-daphnetin (12 mg/L) group and the H-daphnetin+activator (3 μmol/L Src-family activator) group. Cell proliferation was detected by cell counting kit 8 (CCK-8) and colony formation assay. Apoptosis was detected by flow cytometry. The cell migration ability was detected using the scratch healing test. Western blot assay was used to detect the expressions of epithelial-mesenchymal transition-related proteins (E-cadherin, N-cadherin, vimentin), Src and FAK proteins. Results Compared with the control group, the OD₄₅₀ values of 5-8F cells in the L-daphnetin group, the M-daphnetin group and the H-daphnetin group were lower, the number of colony formation was less, the apoptosis rate was higher, the scratch healing rate was lower, the expression of E-cadherin protein was higher, the expressions of N-cadherin and vimentin proteins were lower, and the protein expressions of p-Src and p-FAK were lower (P<0.05), and there were obvious differences between different concentrations (P<0.05). Compared with the H-daphnetin group, the OD₄₅₀ value of 5-8F cells in the H-daphnetin+activator group was higher, the number of colony formation was more, the apoptosis rate was lower, the scratch healing rate was higher, the expression of E-cadherin protein was lower, the expressions of N-cadherin and vimentin proteins were higher, and the protein expressions of p-Src and p-FAK were higher (P<0.05). Conclusion Daphnetin can inhibit the proliferation, migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells by suppressing the Src/FAK pathway.

Key words: Daphnetin, nasopharyngeal carcinoma, genes, src, focal adhesion protein-tyrosine kinases, cell proliferation, cell movement, epithelial-mesenchymal transition

CLC Number:

R739.6

Figures/Tables 10

References 21

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细胞	p-Src/Src	p-FAK/FAK
NP69	0.32±0.05	0.27±0.04
5-8F	0.86±0.09^a	0.82±0.09^a
HNE1	0.74±0.08^a	0.68±0.08^ab
C666-1	0.57±0.07^ab	0.52±0.06^ab
NPC/HK1	0.78±0.09^a	0.70±0.06^ab
F	46.080^＊＊	52.356^＊＊

细胞	p-Src/Src	p-FAK/FAK
NP69	0.32±0.05	0.27±0.04
5-8F	0.86±0.09^a	0.82±0.09^a
HNE1	0.74±0.08^a	0.68±0.08^ab
C666-1	0.57±0.07^ab	0.52±0.06^ab
NPC/HK1	0.78±0.09^a	0.70±0.06^ab
F	46.080^＊＊	52.356^＊＊

组别	OD₄₅₀值	集落形成数/个
对照组	1.96±0.21	152.27±18.39
L-瑞香素组	1.64±0.19^a	126.38±15.87^a
M-瑞香素组	1.32±0.15^ab	99.64±11.92^ab
H-瑞香素组	1.01±0.12^abc	74.46±9.84^abc
H-瑞香素+激活剂组	1.89±0.20^d	148.64±17.16^d
F	30.330^＊＊	29.310^＊＊
组别	凋亡率/%	划痕愈合率/%
对照组	2.59±0.34	41.28±4.83
L-瑞香素组	9.85±1.18^a	33.73±3.96^a
M-瑞香素组	16.57±2.06^ab	26.32±3.21^ab
H-瑞香素组	23.64±2.84^abc	18.95±2.34^abc
H-瑞香素+激活剂组	4.76±0.62^d	39.67±4.65^d
F	158.761^＊＊	34.402^＊＊

组别	OD₄₅₀值	集落形成数/个
对照组	1.96±0.21	152.27±18.39
L-瑞香素组	1.64±0.19^a	126.38±15.87^a
M-瑞香素组	1.32±0.15^ab	99.64±11.92^ab
H-瑞香素组	1.01±0.12^abc	74.46±9.84^abc
H-瑞香素+激活剂组	1.89±0.20^d	148.64±17.16^d
F	30.330^＊＊	29.310^＊＊
组别	凋亡率/%	划痕愈合率/%
对照组	2.59±0.34	41.28±4.83
L-瑞香素组	9.85±1.18^a	33.73±3.96^a
M-瑞香素组	16.57±2.06^ab	26.32±3.21^ab
H-瑞香素组	23.64±2.84^abc	18.95±2.34^abc
H-瑞香素+激活剂组	4.76±0.62^d	39.67±4.65^d
F	158.761^＊＊	34.402^＊＊

组别	E-cadherin	N-cadherin	vimentin
对照组	1.01±0.11	1.06±0.11	1.07±0.12
L-瑞香素组	1.29±0.15^a	0.81±0.09^a	0.83±0.10^a
M-瑞香素组	1.56±0.17^ab	0.56±0.08^ab	0.59±0.07^ab
H-瑞香素组	1.89±0.20^abc	0.31±0.05^abc	0.35±0.05^abc
H-瑞香素+激活剂组	1.09±0.12^d	0.95±0.12^d	0.99±0.13^d
F	33.135^＊＊	63.566^＊＊	54.012^＊＊