Impacts of NGF/TrkA axis on proliferation, apoptosis and invasion of cervical cancer SiHa cells

doi:10.11958/20230318

Abstract

Abstract:

Objective To investigate the impact of nerve growth factor (NGF)/tropomyosin receptor kinase A (TrkA) axis on the proliferation, apoptosis and invasion of cervical cancer SiHa cells. Methods Normal cervical epithelial cells HUCEC and cervical cancer cells SiHa were cultured in vitro. SiHa cells were grouped into the control group (normal culture), the L-NGF group (50 μg/L recombinant human NGF protein), the H-NGF group (100 μg/L recombinant human NGF protein), the H-NGF+L-K252a group (100 μg/L recombinant human NGF protein+50 μg/L K252a) and H-NGF+H-K252a group (100 μg/L recombinant human NGF protein+100 μg/L K252a). The protein expression levels of NGF, TrkA, E-cadherin, N-cadherin and Vimentin were detected by Western blot assay. The proliferation of SiHa cells was detected by CCK-8 method. Apoptosis of SiHa cells was detected by flow cytometry. Cell migration was determined by scratch healing test. Cell invasion was measured by Transwell assay. Results The levels of NGF and TrkA were obviously higher in SiHa cells than those in HUCEC cells (P<0.01). Compared with the control group, levels of NGF and TrkA, proliferative activity, migration rate, number of invasive cells and N-cadherin and Vimentin proteins were obviously higher in the L-NGF group and the H-NGF group (P<0.05), and the apoptosis rate and the level of E-cadherin protein were obviously lower (P<0.05). The above indexes were more obviously different in the H-NGF group than those of the L-NGF group (P<0.05). Compared with the H-NGF group, levels of NGF, TrkA, proliferative activity, migration rate, number of invasive cells and N-cadherin and Vimentin proteins were obviously lower in the H-NGF+L-K252a group and the H-NGF+H-K252a group (P<0.05), and the apoptosis rate and the level of E-cadherin protein were obviously higher (P<0.05). The above indexes in the H-NGF+H-K252a group were more obviously different than those in the H-NGF+L-K252a group (P<0.05). Conclusion Down-regulation of NGF/TrkA axis can inhibit the proliferation and invasion of cervical cancer SiHa cells and promote their apoptosis.

Key words: nerve growth factor, receptor, TrkA, uterine cervical neoplasms, cell proliferation, cell apoptosis, neoplasm invasiveness

CLC Number:

R737.3，R349.54

Figures/Tables 12

References 15

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细胞	NGF	TrkA
HUCEC	0.70±0.06	0.38±0.04
SiHa	1.32±0.17	1.05±0.13
t	8.424^＊＊	12.066^＊＊

细胞	NGF	TrkA
HUCEC	0.70±0.06	0.38±0.04
SiHa	1.32±0.17	1.05±0.13
t	8.424^＊＊	12.066^＊＊

组别	NGF	TrkA
对照组	1.20±0.14	1.02±0.09
L-NGF组	1.56±0.16^a	1.29±0.11^a
H-NGF组	1.89±0.24^ab	1.60±0.14^ab
H-NGF+L-K252a组	1.58±0.18^c	1.28±0.14^c
H-NGF+H-K252a组	1.25±0.16^cd	1.06±0.13^cd
F	14.670^＊＊	21.035^＊＊

组别	NGF	TrkA
对照组	1.20±0.14	1.02±0.09
L-NGF组	1.56±0.16^a	1.29±0.11^a
H-NGF组	1.89±0.24^ab	1.60±0.14^ab
H-NGF+L-K252a组	1.58±0.18^c	1.28±0.14^c
H-NGF+H-K252a组	1.25±0.16^cd	1.06±0.13^cd
F	14.670^＊＊	21.035^＊＊

组别		4 h	24 h	48 h
对照组		0.15±0.01	0.20±0.03	0.31±0.05
L-NGF组		0.16±0.02	0.35±0.04^a	0.45±0.06^a
H-NGF组		0.15±0.01	0.49±0.06^ab	0.65±0.08^ab
H-NGF+L-K252a组		0.14±0.01	0.39±0.02^c	0.52±0.03^c
H-NGF+H-K252a组		0.15±0.02	0.28±0.01^cd	0.37±0.01^cd
F		1.364	54.864^＊＊	39.111^＊＊
组别	72 h		96 h	120 h
对照组	0.40±0.05		0.77±0.05	0.90±0.05
L-NGF组	0.60±0.07^a		1.01±0.12^a	1.18±0.12^a
H-NGF组	0.78±0.09^ab		1.28±0.09^ab	1.45±0.09^ab
H-NGF+L-K252a组	0.63±0.04^c		1.07±0.09^c	1.24±0.15^c
H-NGF+H-K252a组	0.44±0.02^cd		0.86±0.04^cd	0.99±0.10^cd
F	40.457^＊＊		33.844^＊＊	24.402^＊＊