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

Tab.1

Fig.1

Tab.2

Fig.2

Tab.3

Tab.4

Fig. 3

Fig. 4

Fig.5

Tab.5

Tab.6

Fig. 6

References 22

1	YANG H P， GUO Y， WANG D W， et al. Effect of TAK1 on osteogenic differentiation of mesenchymal stem cells by regulating BMP-2 via Wnt/β-catenin and MAPK pathway［J］. Organogenesis， 2018， 14（1）：36-45.
2	HUANG F T， PENG J F， CHENG W J， et al. MiR-143 targeting TAK1 attenuates pancreatic ductal adenocarcinoma progression via MAPK and NF-κB pathway in vitro［J］. Dig Dis Sci， 2017， 62（4）：944-957.
3	廉亮亮，齐书山，袁洪志. miR-143-3p通过靶向TAK1抑制肺癌增殖和侵袭［J］. 生物技术通讯， 2019， 30（3）：332-337.
4	曹赛，程梅容，刘素娥，等. 食管癌组织中TAK1和TAB1的表达及与临床预后的相关性［J］. 南方医科大学学报， 2018， 38（7）：895-900.
5	LIN P F， NIU W B， PENG C， et al. The role of TAK1 expression in thyroid cancer［J］. Int J Clin Exp Pathol， 2015， 8（11）：14449-14456.
6	CUI D J， WU Y， WEN D H. CD34， PCNA and CK19 expressions in AFP- hepatocellular carcinoma［J］. Eur Rev Med Pharmacol Sci， 2018， 22（16）：5200-5205.
7	WANG X H， HU Y D， CUI J Y， et al. Coordinated targeting of MMP-2/MMP-9 by miR-296-3p/FOXCUT exerts tumor-suppressing effects in choroidal malignant melanoma［J］. Mol Cell Biochem， 2018， 445（1/2）：25-33.
8	CI Y Q， ZHANG Y B， LIU Y J， et al. Myricetin suppresses breast cancer metastasis through down-regulating the activity of matrix metalloproteinase （MMP）-2/9［J］. Phytother Res， 2018， 32（7）：1373-1381.
9	DALAKOURAS A， WASSENEGGER M， DADAMI E，et al. Genetically modified organism-free RNA interference： exogenous application of RNA molecules in plants［J］. Plant Physiol， 2020， 182（1）：38-50.
10	GUTBROD M J， MARTIENSSEN R A. Conserved chromosomal functions of RNA interference［J］. Nat Rev Genet， 2020， 21（5）：311-331.
11	BRÜGGENWIRTH I M A， MARTINS P N. RNA interference therapeutics in organ transplantation： The dawn of a new era［J］. Am J Transplant， 2020， 20（4）：931-941.
12	YANG L D， JOSEPH S， SUN T L， et al. TAK1 regulates endothelial cell necroptosis and tumor metastasis［J］. Cell Death Differ， 2019， 26（10）：1987-1997.
13	YUNG M M， TANG H W， CAI P C， et al. GRO-α and IL-8 enhance ovarian cancer metastatic potential via the CXCR2-mediated TAK1/NFκB signaling cascade［J］. Theranostics， 2018，8（5）：1270-1285.
14	CHEN J M， ZHANG N， WEN J M， et al. Silencing TAK1 alters gene expression signatures in bladder cancer cells［J］. Oncol Lett， 2017， 13（5）：2975-2981.
15	JIAO H L， YE Y P， YANG R W， et al. Downregulation of SAFB Sustains the NF-κB pathway by targeting TAK1 during the progression of colorectal cancer［J］. Clin Cancer Res， 2017，23（22）：7108-7118.
16	TOTZKE J， GURBANI D， RAPHEMOT R， et al. Takinib， a selective TAK1 inhibitor， broadens the therapeutic efficacy of TNF-α inhibition for cancer and autoimmune disease［J］. Cell Chem Biol， 2017， 24（8）：1029-1039.e7.
17	GUAN S， LU J X， ZHAO Y L， et al. TAK1 inhibitor 5Z-7-oxozeaenol sensitizes cervical cancer to doxorubicin-induced apoptosis［J］. Oncotarget， 2017， 8（20）：33666-33675.
18	BAN Z F， HE J N， TANG Z Z， et al. LRG-1 enhances the migration of thyroid carcinoma cells through promotion of the epithelial-mesenchymal transition by activating MAPK/p38 signaling［J］. Oncol Rep， 2019， 41（6）：3270-3280.
19	HAN S E， PARK C H， NAM-GOONG I S， et al. Anticancer effects of baicalein in FRO thyroid cancer cells through the up-regulation of ERK/p38 MAPK and Akt pathway［J］. In Vivo， 2019， 33（2）：375-382.
20	WU L P， CAO Z Y， JI L， et al. Loss of TRADD attenuates pressure overload-induced cardiac hypertrophy through regulating TAK1/P38 MAPK signalling in mice［J］.Biochem Biophys Res Commun，2017， 483（2）：810-815.
21	FAN Y W， LI M Y， MA K， et al. Dual-target MDM2/MDMX inhibitor increases the sensitization of doxorubicin and inhibits migration and invasion abilities of triple-negative breast cancer cells through activation of TAB1/TAK1/p38 MAPK pathway［J］. Cancer Biol Ther， 2019， 20（5）：617-632.
22	苏书娟. 牡荆葡基黄酮调节TAK1/AMPK通路增强人肝癌细胞的放射敏感性［J］. 中国免疫学杂志， 2018， 34（8）：1158-1162.

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

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 12

References 22

Related Articles 15

Metrics

Comments

Recommended 10

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

[1]	Xiao ZHOU,Haitao DAI,Yunshan DING,Linjie ZHANG,Nan WU. Network pharmacology analysis and in vitro experimental validation of anticancer effect of grape seed proanthocyanidins against oral squamous cell carcinoma [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 651-662.
[2]	Qiaotong REN,Weihua WU,Xingxiang WANG,Shichao WANG,Yipeng CHENG,Chun LI. Effect of interleukin-17 recepter B gene silencing on proliferation, invasion and apoptosis of lung adenocarcinoma A549 cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 738-745.
[3]	Yao ZHENG,Mingxia FU,Weichen WANG,Weiwei CHEN,Yuchen HAN,Yu BAI,Jiajia AN. Effect of sodium selenite on biological behaviors of breast cancer doxorubicin-resistant MCF-7/ADR cells [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 410-417.
[4]	Li ZHAI,Meng CHEN,Jianbo LUO,Aili ZHANG,Liangxiao WANG,Ying WEI,Xi ZHANG. Targeting relationship between miR-214-3p and EZH2 and its effects on proliferation， invasion， and apoptosis of ovarian cancer SKOV3 cells [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 460-468.
[5]	Hong LI,Panpan YU,Zouyu ZHAO,Chongfeng SUN,Hui QIAO,Ping YANG. Expression of GSTO1 in ovarian cancer tissue and effect of N-glycosylation site mutations on biological behaviors of epithelial ovarian cancer cells [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 469-482.
[6]	Yunshan DING,Haitao DAI,Min CHEN,Xiaohui HAO,Xiao ZHOU,Nan WU. Effect of silencing TRPV2 gene and cannabidiol on biological behaviors of oral squamous cell carcinoma CAL-27 cells [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 143-151.
[7]	Yan ZHAO,Huawei WU. Inhibitory effect of inhibition of miR-17-5p on proliferation of spinal cord astrocytes of rats induced by scratch injury through targeting Mfn2 expression [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 81-92.
[8]	Xiaohan YAO,Mingchen YAO,Zhiqing WANG,Heyang LI,Yan YAN,Ningjing LEI. Effect of silencing GPR139 gene on proliferation, apoptosis and autophagy of breast cancer cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 1-9.
[9]	Sifan FENG,Yunfeng LI,Jiaying WANG,Fubin MA,Yan WANG. Effects of heme-binding protein 1 gene knockdown on proliferation, migration, and inflammatory response of microglia BV2 and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2025, 51(6): 1532-1541.
[10]	Lulu FU,Yinggang ZOU,Xiaoyu ZHENG,Xueying ZHANG,Jingshun ZHANG,Min WANG,Qiang ZHANG,Lianwen ZHENG. Expression of placenta expressed transcription factor 1 in ovarian tissue of polycystic ovary syndrome rats and its effect on proliferation of rat ovarian granulosa cells [J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1177-1184.
[11]	Pingsheng ZHU,Sitang GE,Lugen ZUO,Deli CHEN,Yangyang ZHANG. Effect of miR-325-3p targeting PRELID1 gene in regulation of EMT pathway on invasion and migration of colon cancer cells and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1185-1193.
[12]	Wenxuan LI,Minru ZONG. Research progress in role of migration of Schwann cells in repairment of peripheral nerve injury [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 1137-1144.
[13]	Zhongjun SHEN,Yao ZHAO,Mingbo JIA,Liyan ZHAO. Research progress in effects of hypoxia-inducible factors on cell migration and invasion during epithelial-mesenchymal transition in glioma cells [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 1145-1154.
[14]	Yixuan GAO,Peng WANG,Silong ZHANG,Ruijuan GAO,Yingfang MA,Keke ZHANG,Dan FENG,Zongqi HUANG,Ketao MA,Li LI,Junqiang SI. Inhibitory effect of safranal on proliferation, migration and phenotypic transformation of vascular smooth muscle cells of rats induced by high glucose in vitro [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 948-957.
[15]	Xiaoshuang HE,Lina XU,Mei CUI,Yu ZHAO,Bei WANG,Zheng HUANG,Yuchao WANG,Wenyan XIN,Chao WU. Effects of lncRNA DUXAP8 in lung cancer A549 cells-derived exosomes on lung cancer cell growth and its mechnism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 958-967.