Effects of cordycepin on proliferation， apoptosis and epithelial mesenchymal transition in nasopharyngeal carcinoma cells by regulating Akt/GSK-3β/Snail signaling pathway

doi:10.3760/cma.j.cn371439-20250415-00010

Abstract

Abstract:

Objective To explore the effects of cordycepin on proliferation， apoptosis and epithelial mesenchymal transition （EMT） in nasopharyngeal carcinoma cells by regulating Akt/GSK-3β/Snail signaling pathway. Methods The nasopharyngeal carcinoma CNE-1 cells were divided into low， medium， high dose cordycepin groups （5， 10， 20 μmol/L cordycepin）， high dose cordycepin+SC79 group （20 μmol/L cordycepin+10 μmol/L Akt/GSK-3β/Snail signaling pathway activator SC79） and control group， and cells were intervened for 24 h in each treatment group. Cell cloning situation was detected by plate cloning assay； cell migration was detected by cell scratch assay； Transwell chamber method was used to detect cell invasion； flow cytometry was used to detect cell apoptosis； Western blotting was used to detect the expression of Akt， p-Akt， GSK-3β， p-GSK-3β， Snail， N-cadherin， vimentin， E-cadherin， Bcl-2 and Bax proteins. Results The results of the plate cloning assay showed that the numbers of cell clones in the control group， the low， medium and high dose cordycepin groups， and the high dose cordycepin+SC79 group were 116.83±4.35， 93.17±4.01， 70.50±3.83， 52.33±3.41， and 69.17±3.29， respectively. The results of the cell scratch assay showed that the scratch healing rates of the 5 groups of cells were （38.16±2.37）%，（30.27±2.26）%，（24.19±2.01）%，（17.15±1.92）%， and （22.73±2.15）%. The results of the Transwell method showed that the numbers of cell invasions in the 5 groups were 96.50±3.49， 78.33±3.12， 62.17±2.93， 43.50±2.58， and 57.33±2.27， respectively. The results of flow cytometry showed that the apoptosis rates of the 5 groups of cells were （2.46±0.35）%，（15.93±1.59）%，（23.17±1.64）%，（29.39±1.86）%， and （24.84±1.72）%， respectively， all with statistically significant differences （F=260.21， P<0.001； F=83.58， P<0.001； F=295.35， P<0.001； F=282.13， P<0.001）. The numbers of cell clones and the numbers of cell invasions in the control group and the low， medium and high dose cordycepin groups decreased successively， and the scratch healing rates decreased successively， while the cell apoptosis rates increased successively （all P<0.05）. Compared to the high dose cordycepin group， the high dose cordycepin+SC79 group had more cell clones and cell invasions， a higher scratch healing rate， and a lower apoptosis rate （all P<0.05）. The results of Western blotting showed that the expression levels of N-cadherin protein in the control group， the low， medium and high dose cordycepin groups， and the high dose cordycepin+SC79 group were 0.82±0.08， 0.68±0.05， 0.55±0.04， 0.39±0.02 and 0.53±0.06， respectively. The expression levels of vimentin protein in the 5 groups were 0.94±0.09， 0.76±0.07， 0.59±0.06， 0.42±0.03 and 0.56±0.05， respectively. The expression levels of E-cadherin protein in the 5 groups were 0.42±0.04， 0.57±0.06， 0.73±0.07， 0.91±0.09 and 0.78±0.07， respectively. The expression levels of Bcl-2 protein in the 5 groups were 0.89±0.08， 0.73±0.06， 0.57±0.05， 0.43±0.03 and 0.55±0.07， respectively. The expression levels of Bax protein in the 5 groups were 0.31±0.03， 0.45±0.05， 0.61±0.06， 0.78±0.08 and 0.67±0.06， respectively. The expression levels of p-Akt/Akt in the 5 groups were 0.87±0.08， 0.75±0.06， 0.60±0.05， 0.46±0.03 and 0.58±0.04， respectively. The expression levels of p-GSK-3β/GSK-3β in the 5 groups were 0.98±0.09， 0.84±0.08， 0.69±0.06， 0.54±0.05 and 0.66±0.06， respectively. The expression levels of Snail protein in the 5 groups were 0.92±0.08， 0.74±0.05， 0.56±0.05， 0.37±0.03 and 0.52±0.06， respectively， all with statistically significant differences （F=54.89， P<0.001； F=60.27， P<0.001； F=47.10， P<0.001； F=52.26， P<0.001； F=60.67， P<0.001； F=50.94， P<0.001； F=36.10， P<0.001； F=85.13， P<0.001）. In the control group and the low， medium and high dose cordycepin groups， the expression levels of N-cadherin， vimentin， Bcl-2， p-Akt/Akt， p-GSK-3β/GSK-3β， and Snail protein decreased successively （all P<0.05）， while the expression levels of E-cadherin and Bax protein increased successively （all P<0.05）. Compared with the high dose cordycepin group， the expression levels of N-cadherin， vimentin， Bcl-2， p-Akt/Akt， p-GSK-3β/GSK-3β， and Snail protein in the high dose cordycepin+SC79 group were higher （all P<0.05）， while the expression levels of E-cadherin and Bax protein were lower （both P<0.05）. Conclusions Cordycepin can promote apoptosis of nasopharyngeal carcinoma cells and inhibit cell invasion， migration， proliferation and EMT， which may be related to the down-regulation of Akt/GSK-3β/Snail signaling pathway.

Key words: Nasopharyngeal neoplasms, Cordycepin, Akt/GSK-3β/Snail signaling pathway, Epithelial mesenchymal transition

Shi Rui, Dai Jian, Chen Ran, Hu Lili. Effects of cordycepin on proliferation， apoptosis and epithelial mesenchymal transition in nasopharyngeal carcinoma cells by regulating Akt/GSK-3β/Snail signaling pathway[J]. Journal of International Oncology, 2026, 53(2): 65-72.

Figures/Tables 10

References 25

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虫草素浓度（μmol/L）	CNE-1细胞	5-8F细胞	NP69细胞
0	98.16±3.95	98.89±4.18	97.27±3.86
1	93.43±3.67	97.26±4.02	96.02±3.79
2	89.58±3.57^a	95.12±3.86	96.59±3.81
5	83.94±3.26^ab	90.37±3.79^ab	94.64±3.26
10	71.85±3.01^abcd	79.58±3.54^abcd	93.78±3.56
20	62.72±2.84^abcde	67.64±3.37^abcde	94.35±3.17
40	50.79±2.61^abcdef	59.49±3.22^abcdef	93.71±3.25
80	37.73±2.72^abcdefg	51.13±3.01^abcdefg	93.19±3.42
F值	269.03	154.81	1.08
P值	<0.001	<0.001	0.393

组别	细胞划痕愈合率（%）	细胞侵袭个数（个）
对照组	38.16±2.37	96.50±3.49
虫草素低剂量组	30.27±2.26^a	78.33±3.12^a
虫草素中剂量组	24.19±2.01^ab	62.17±2.93^ab
虫草素高剂量组	17.15±1.92^abc	43.50±2.58^abc
虫草素高剂量+SC79组	22.73±2.15^d	57.33±2.27^d
F值	83.58	295.35
P值	<0.001	<0.001

组别	神经钙黏素	波形蛋白	上皮钙黏素	Bcl-2	Bax
对照组	0.82±0.08	0.94±0.09	0.42±0.04	0.89±0.08	0.31±0.03
虫草素低剂量组	0.68±0.05^a	0.76±0.07^a	0.57±0.06^a	0.73±0.06^a	0.45±0.05^a
虫草素中剂量组	0.55±0.04^ab	0.59±0.06^ab	0.73±0.07^ab	0.57±0.05^ab	0.61±0.06^ab
虫草素高剂量组	0.39±0.02^abc	0.42±0.03^abc	0.91±0.09^abc	0.43±0.03^abc	0.78±0.08^abc
虫草高剂量+SC79组	0.53±0.06^d	0.56±0.05^d	0.78±0.07^d	0.55±0.07^d	0.67±0.06^d
F值	54.89	60.27	47.10	52.26	60.67
P值	<0.001	<0.001	<0.001	<0.001	<0.001

组别	p-Akt/Akt	p-GSK-3β/GSK-3β	Snail
对照组	0.87±0.08	0.98±0.09	0.92±0.08
虫草素低剂量组	0.75±0.06^a	0.84±0.08^a	0.74±0.05^a
虫草素中剂量组	0.60±0.05^ab	0.69±0.06^ab	0.56±0.05^ab
虫草素高剂量组	0.46±0.03^abc	0.54±0.05^abc	0.37±0.03^abc
虫草素高剂量+ SC79组	0.58±0.04^d	0.66±0.06^d	0.52±0.06^d
F值	50.94	36.10	85.13
P值	<0.001	<0.001	<0.001