IL-17A在非小细胞肺癌组织中的表达及其通过NF-κB信号通路对VEGF表达的调控作用

doi:10.13481/j.1671-587X.20220421

Abstract

Abstract: Objective

To investigate the role of interleukin-17A （IL-17A） in the occurrence and development of non-small cell lung cancer （NSCLC） and its regulation on vascular endothelial growth factor （VEGF） through the nuclear factor-κB （NF-κB） signaling pathway， and to elucidate their related mechanisms.

Methods

A total of fifty-five postoperative paraffin-embedded specimens of the patients with NSCLC were collected. The expressions of IL-17A and CD31 in normal lung tissue and NSCLC tissue were detected by immunohistochemical staining， the correlations between the positive expression rate of IL-17A and the clinicopathological parameters of NSCLC were analyzed， and the correlation between the expressions of IL-17A and CD31-marked microvessel density （MVD） was analyzed by Pearson correlation analysis. The human lung adenocarcinoma A549 cells at logarithmic growth stage were selected and divided into control group （A549 cells）， exogenous recombinant human IL-17A （rhIL-17A） group， BAY11-7082 （NF-κB signaling pathway inhibitor） group and combination group （rhIL-17A+BAY11-7082）. Western blotting method was performed to detect the expression levels of IL-17 receptor A （IL-17RA）， P65，p-P65 and VEGF proteins in the cells in various groups. The MTT method was used to detect the proliferation abilities of human umbilical vein endothelial cells （HUVECs） in various groups after treated with the A549 cell culture supernatant for 48 h.

Results

The positive expression rate of IL-17A in NSCLC tissue was significantly higher than that in normal lung tissue （P<0.05），the positive expression rate of IL-17A in poor-differentiation group was significantly higher than that in well-moderate differentiation group （P<0.05）， and the expression of IL-17A was positively correlated with the MVD in NSCLC tissue （r=0.329， P<0.05）.Compared with control group， the expression levels of IL-17RA， p-P65 and VEGF proteins in the cells in rhIL-17A group were increased （P<0.05）， and the expression levels of IL-17RA， p-P65 and VEGF proteins in the cells in BAY11-7082 group were decreased （P<0.01）. Compared with rhIL-17A group， the expression levels of IL-17RA， p-P65 and VEGF proteins in the A549 cells in combination group were significantly decreased （P<0.05）. After treated with A549 cell culture supernatant， compared with control group， the proliferation ability of the HUVEC in rhIL-17A group was increased （P<0.05），and the proliferation ability in the cells in BAY11-7082 group was decreased （P<0.05）. Compared with the rhIL-17A group， the proliferation ability of the HUVECs in combination group was significantly decreased （P<0.05）.

Conclusion

IL-17A is highly expressed in poorly differentiated NSCLC cells， and it can promote the proliferation of HUVECs by regulating the expression of VEGF via NF-κB signaling pathway.

Key words: Interleukin-17A, Non-small cell lung cancer, Nuclear transcription factor-kappa B, Vascular endothelial growth factor, Human umbilical vein endothelial cells

CLC Number:

R734.2

Chengyuan HE,Hongyu YANG,Yujing TAN,Hang SU,Hongshu LI,Chun LI. Expression of IL-17A in non-small cell lung cancer tissue and its regulation on VEGF expression via NF-κB signaling pathway[J].Journal of Jilin University(Medicine Edition), 2022, 48(4): 1003-1009.

Figures/Tables 9

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References 26

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Clinicopathological feature	n	IL-17A		P
Clinicopathological feature	n	Positive	Negative	P
Age(year)
≥60	28	15（53.6）	13（46.4）	0.322
<60	27	18（66.7）	9（33.3）
Sex
Male	35	20（57.1）	15（42.9）	0.567
Female	20	13（65.0）	7（35.0）
Tumor size
≤3 cm	24	16（66.7）	8（33.3）	0.375
>3 cm	31	17（54.8）	14（45.2）
Histology
Squamous carcinoma	35	20（57.1）	15（42.9）	0.567
Adenocarcinoma	20	13（65.0）	7（35.0）
Differentiation
Well- moderate	32	15（43.8）	17（56.2）	0.019
Poor	23	18（78.3）	5（21.7）
Lymph node metastasis
Yes	24	16（66.7）	8（33.3）	0.375
No	31	17（54.8）	14（45.2）
TNM staging
Ⅰ	27	14（51.9）	13（48.1）	0.226
Ⅱ+Ⅲ	28	19（67.9）	9（32.1）

Group	IL-17RA	p-P65	VEGF
Control	0.36±0.07	0.54±0.06	0.23±0.04
rhIL-17A	0.66±0.06 ^*	0.76±0.07 ^*	0.37±0.03 ^*

Group	IL-17RA	p-P65	VEGF
Control	0.77±0.08	1.25±0.08	0.26±0.03
rhIL-17A	0.99±0.06 ^*	1.53±0.08 ^*	0.37±0.04 ^*
BAY11-7082	0.12±0.02 ^**	0.52±0.07 ^**	0.15±0.01 ^**
Combination	0.45±0.08^△	1.01±0.07^△	0.24±0.02^△
F	63.61	59.09	30.345
P	<0.01	<0.01	<0.01

Group	Proliferation activity
Control	0.887±0.035
rhIL-17A	0.974±0.036^*
BAY11-7082	0.750±0.100^*
Combination	0.851±0.057^△

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Expression of IL-17A in non-small cell lung cancer tissue and its regulation on VEGF expression via NF-κB signaling pathway

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MVD	IL-17A expression		Total	r	P
MVD	Positive	Negative	Total	r	P
High	23	8	31	0.329	0.014
Low	10	14	24
Total	33	22	55