siRNA 靶向沉默TAK1基因对甲状腺癌细胞增殖、迁移和p38 MAPK信号通路的抑制作用

doi:10.13481/j.1671-587x.20210115

Abstract

Abstract: Objective

To investigate the effects of siRNA targeting silencing transforming growth factor β-activated kinase 1 （TAK1） gene on the proliferation，migration and p38 mitogen-activated protein kinase （MAPK） signaling pathway of thyroid cancer cells，and to clarify the possible mechanism of silencing TAK1 gene in thyroid cancer.

Methods

Quantitative real-time polymerase chain reaction （qRT-PCR） and Western blotting method were used to determine the expression levels of TAK1 mRNA and proteins in the thyroid cancer 8505C， NPA， BCPAP and KMH-2 cells. The KMH-2 cells were randomly divided into blank control group， negative control group and siRNA-TAK1 group. The cells in blank control group were not transfected，the cells in negative control group were transfected with negative control siRNA，and the cells in siRNA-TAK1 group were transfected with TAK1 siRNA. MTT method was used to measure the cell proliferation activity. Transwell assay was used to measure the invasion and migration abilities of cells. Western blotting method was used to determine the expression levels of proliferating cell nuclear antigen （PCNA）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9（MMP-9），p38 and phosphorylated p38 （p-p38） proteins in the cells in various groups.

Results

Compared with the normal thyroid epithelial cells Nthyori3-1， the expression levels of TAK1 mRNA and protein in thyroid cancer 8505C， NPA， BCPAP and KMH-2 cells were significantly increased（P<0.05）. Compared with blank control group and negative control group， the expression levels of TAK1 mRNA and protein in the KMH-2 cells in siRNA-TAK1 group were reduced （P<0.05）， the cell proliferation activities were reduced （P<0.05）， the number of invasion cells and migration cells was reduced （P<0.05），and the expression levels of PCNA， MMP-2， MMP-9 and p-p38 proteins in the cells were decreased （P<0.05）. There were no significant differences in the indexes mentioned above of the KMH-2 cells between blank control group and negative control group（P>0.05）.

Conclusion

SiRNA targeting silencing TAK1 gene can inhibit the proliferation，invasion and migration of thyroid cancer cells through inhibiting the activation of p38 MAPK signaling pathway.

Key words: siRNA, transforming growth factor β-activated kinase 1, thyroid neoplasms, cell proliferation, cell migration, p38 MAPK

CLC Number:

R736.1

Chunying ZHANG,Guangwei YIN,Mingda YOU,Hong CHEN,Yaojie HU,Yanbing LI,Chunyou CHEN. Inhibitory effects of siRNA targeting silencing TAK1 gene on proliferation and migration of thyroid cancer cells and p38 MAPK signaling pathway[J].Journal of Jilin University(Medicine Edition), 2021, 47(1): 110-117.

Figures/Tables 12

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References 22

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Cell	TAK1 mRNA	TAK1 protein
Nthyori3-1	1.00±0.12	0.12±0.04
8505C	1.68±0.11 ^*	0.48±0.06 ^*
NPA	1.97±0.13 ^*	0.82±0.08 ^*
BCPAP	1.84±0.15 ^*	0.78±0.07 ^*
KMH-2	2.32±0.16 ^*	0.94±0.10 ^*
F	90.663	144.385
P	<0.01	<0.01

Group	TAK1 mRNA	TAK1 protein
Black control	1.00±0.08	0.92±0.13
Negative control	0.98±0.09	0.94±0.12
siRNA-TAK1	0.41±0.06^*△	0.26±0.07^*△
F	130.215	86.862
P	<0.01	<0.01

Group	Proliferation activity
Group	(t/h) 24	48	72
Blank control	0.412±0.033	0.784±0.041	1.273±0.044
Negative control	0.415±0.030	0.779±0.043	1.268±0.046
siRNA-TAK1	0.327±0.025^*△	0.461±0.038^*△	0.691±0.038^*△
F	20.055	144.586	427.743
P	<0.01	<0.01	<0.01

Group	Number of invasion cells	Number of migration cells
Blank control	378.46±37.35	215.68±28.49
Negative control	382.19±41.26	214.94±31.02
siRNA-TAK1	153.26±35.49^*△	92.13±25.18^*△
F	82.852	44.111
P	<0.01	<0.01

Group	PCNA	MMP-2	MMP-9
Blank control	0.43±0.08	0.28±0.06	0.33±0.05
Negative control	0.45±0.09	0.30±0.07	0.34±0.06
siRNA-TAK1	0.21±0.05^*△	0.13±0.03^*△	0.15±0.04^*△
F	21.906	19.287	31.182
P	<0.01	<0.01	<0.01

Inhibitory effects of siRNA targeting silencing TAK1 gene on proliferation and migration of thyroid cancer cells and p38 MAPK signaling pathway

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Group	p38	p-p38
Blank control	0.59±0.09	0.28±0.05
Negative control	0.57±0.10	0.29±0.06
siRNA-TAK1	0.62±0.12	0.10±0.02^*△
F	0.409	36.938
P	0.670	<0.01

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