Notch1信号通路对肺腺癌细胞侵袭和顺铂耐药的影响及其机制

doi:10.13481/j.1671-587X.20210513

Abstract

Abstract:

Objective： To observe the effects of blocking Notch1 signaling pathway on the cell invasion， angiogenesis， epithelial mesenchymal transition （EMT），and chemosensitivity to cisplatin of the lung adenocarcinoma cells， and to elucidate the role and related mechanism of Notch1 signaling pathway in the cell invasion， metastasis and cisplatin resistance of lung adenocarcinoma.

Methods

The A549 cells were treated with DAPT （N-［N-（3，5-Difluorophenacetyl）-l-alanyl］-S-phenylglycine t-butyl ester） as an inhibitor of Notch1 signaling pathway and divided into 0 μmol·L^－1 DAPT group（control group），10 μmol·L^－1 DAPT group and 20 μmol·L^－1 DAPT group. Transwell chamber assay was used to detect the number of invasion cells of A549 cells in various groups； ELISA assay was used to detect the levels of matrix metalloproteinase-9 （MMP-9） and vascular endothelial growth factor （VEGF） in the supernatant of cells in various groups； Western blotting method was performed to detect the expression levels of Notch1， Hes1， Vimentin， N-cadherin and E-cadherin proteins in the cells in various groups； CCK-8 method was used to detect the inhibitory rates of growth of the A549 cells after treated with cisplatin （0.625－5.000 mg·L^－1） in various groups； Western blotting method was used to detect the expression levels of Notch1 and Hes1 proteins in the A549 cells after treated with 0 and 1.250 mg·L^－1cisplatin alone， and the expression levels of P-glycoprotein （P-gp） and multidrug resistance-associated protein 1 （MRP1） proteins in the A549 cells after treated with 1.250 mg·L^－1cisplatin alone or combination with 10 μmol·L^－1 DAPT.

Results

Compared with control group， the number of invasion cells in 10 and 20 μmol·L^－1 DAPT groups was significantly decreased （P<0.05 or P<0.01），the levels of MMP-9 and VEGF in in the cell supernatant were significantly decreased （P<0.05 or P<0.01）， the expression levels of Notch1， Hes1， Vimentin and N-cadherin proteins in the A549 cells were significantly decreased （P<0.05 or P<0.01），and the expression levels of E-cadherin protein were significantly increased （P<0.05）.Compared with control group， the inhibitory rates of growth of the A549 cells in 10 and 20 μmol·L^－1 DAPT groups were significantly increased after treated with different concentrations of cisplatin （P<0.05 or P<0.01）.Compared with 0 mg·L^－1cisplatin group， the expression levels of Notch1， Hes1， P-gp and MRP1 proteins in the A549 cells in 1.250 mg·L^－1cisplatin group were significantly increased （P<0.05 or P<0.01）. Compared with 1.250 mg·L^－1cisplatin group， the expression levels of P-gp and MRP1 proteins in the A549 cells in 1.250 mg·L^－1cisplatin combined with 10 μmol·L^－1 DAPT group were significantly decreased （P<0.01）.

Conclusion

Blocking Notch1 signaling pathway may inhibit the cell invasion and reduce the cisplatin resistance in the lung adenocarcinoma cells.

Key words: lung neoplasm, Notch1, cell invasion, cell metastasis, cisplatin, multidrug resistance

CLC Number:

R734.2

Lisong CHEN,Hui FANG,Hefei WANG,Chengyuan HE,Xiaodong GAI,Chun LI. Effects of Notch1 signaling pathway on invasion and cisplatin resistance of lung adenocarcinoma cells and their mechanisms[J].Journal of Jilin University(Medicine Edition), 2021, 47(5): 1171-1177.

Figures/Tables 8

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Group	MMP-9	VEGF
Control	438.00±23.07	351.90±18.49
10 μmol·L^－1 DAPT	379.17±20.28 ^*	302.83±16.50 ^*
20 μmol·L^－1 DAPT	313.57±17.49 ^**	284.17±14.27 ^**
F	27.919	13.467
P	< 0.01	< 0.01

Group	Notch1	Hes1	Vimentin	N-cadherin	E-cadherin
Control	1.17±0.15	1.01±0.10	0.74±0.11	0.68±0.09	0.56±0.06
10 μmol·L^－1 DAPT	0.77±0.08^*	0.74±0.06^*	0.38±0.07^**	0.48±0.06^**	0.64±0.08
20 μmol·L^－1 DAPT	0.51±0.06^**	0.53±0.06^**	0.30±0.05^**	0.29±0.04^**	0.81±0.11^*
F	28.288	31.269	25.774	35.10	8.808
P	<0.01	<0.05	<0.01	<0.01	<0.05

Group	Inhibitory rate of growth
Group	(mg·L^－1)	0.625	1.250	2.500	5.000
DDP		7.51±0.66	14.30±2.04	24.90±2.72	47.50±2.50
DDP+10 μmol·L^－1 DAPT		9.37±1.12^*	22.30±2.70^*	32.90±2.20^*	53.90±2.10^*
DDP+20 μmol·L^－1 DAPT		12.90±1.90^**	25.60±1.29^**	36.70±2.07^**	55.30±2.52^**
F		12.974	23.253	19.943	9.105
P		<0.01	<0.01	<0.01	<0.05

Group	P-gp	MRP1
0 mg·L^－1 DDP	0.29±0.04	0.35±0.06
1.250 mg·L^－1 DDP	0.57±0.06^*	0.65±0.07^*
1.250 mg·L^－1 DDP+ 10 μmol·L^－1 DAPT	0.35±0.05^△	0.41±0.05^△
F	25.807	22.217
P	< 0.01	< 0.01

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Effects of Notch1 signaling pathway on invasion and cisplatin resistance of lung adenocarcinoma cells and their mechanisms

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