氯化两面针碱通过JAK2/STAT3信号通路对胶质瘤细胞上皮-间质转化的抑制作用

doi:10.13481/j.1671-587x.20210110

Abstract

Abstract: Objective

To investigate the inhibitory effect of nitidine chloride （NC） on the epithelial-mesenchymal transition （EMT） of glioma cells， and to clarify signaling mechanism of the inhibitory effect.

Methods

The human glioma U87 cells cultured in vitro were divided into control group （without treatment），EMT group ［EMT induced by transforming growth factor-β1（TGF-β1）］ and different concentrations （2.5， 5.0， 7.5 and 10 μmol·L^－1） of NC groups （different concentrations of NC+10 μg·L^－1 TGF-β1）. After 48 h of treatment， the cell migration rates，the invasion rates and the expression levels of EMT-related proteins and JAK2 / STAT3 pathway-related proteins were detected by wound healing test， Transwell invasion test and Western blotting method， respectively. When inducing EMT in the U87 cells， the JAK2 / STAT3 pathway was blocked by STAT3 inhibitor WP1066. The U87 cells were divided into control group （without treatment）， EMT group （10 μg·L^－1TGF-β1） and EMT + WP1066 group （10 μg·L^－1 TGF-β1 + 8 μmol·L^－1WP1066）； the expression levels of EMT-related proteins in the cells in various group were detected by Western blotting method.

Results

The cell migration rate and invasion rate in EMT group were significantly higher than those in control group （P<0.01）， while the cell migration rate and invasion rate in NC group were significantly lower than those in EMT group （P<0.01）. Compared with control group， the expression level of E-cadherin in the U87 cells in EMT group was decreased（P<0.01）， while the expression levels of N-cadherin， vimentin， β-catenin， and the transcription factors Snail， Slug and Twist1 proteins were increased （P<0.05 or P<0.01）. Compared with EMT group， the expression levels of E-cadherin in different concentrations of NC groups were increased （P<0.05 or P<0.01）， and the expression levels of N-cadherin， vimentin， β-catenin， and transcription factors Snail， Slug and Twist1 proteins were significantly reduced （P<0.01） in a concentration-dependent manner； 7.5 μmol·L^－1NC could significantly inhibit the EMT of glioma cells. Compared with control group，the expression levels of JAK2 and p-JAK2 proteins in the cells in EMT group had no significant differences（P>0.05），and the expression levels of STAT3 and p-STAT3 in the cells were significantly decreased（P<0.05）.Compared with EMT group，the expression levels of JAK2，p-JAK2， STAT3 and p-STAT3 proteins in the cells in different NC groups were significantly reduced （P<0.01）. In JAK2/STAT3 pathway blocking experiment by STAT3 inhibitor WP1066， the expression levels of E-cadherin proteins in the cells in EMT+WP1006 group was significantly increased（P<0.01），and the expression levels of N-cadherin，vimentin，β-catein Twist1，Slug and Snail proeins were significantly decreased（P<0.01）.

Conclusion

NC can inhibit the EMT of glioma cells， and the inhibitory effect is related to JAK2/STAT3 signaling pathway.

Key words: glioma cells, nitidine chloride, epithelial-mesenchymal transition, Janus kinase signal transducers 2/signal transducer and activator of transcription 3 signaling pathway

CLC Number:

R739.41

Mingbo JIA,Ying SUN,Ying WANG,Yanke SONG,Liyan ZHAO. Inhibitory effect of nitidine chloride on epithelial-mesenchymal transition of glioma cells through JAK2/STAT3 signaling pathway[J].Journal of Jilin University(Medicine Edition), 2021, 47(1): 73-81.

Figures/Tables 10

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References 24

1	KAHLERT U D， NIKKHAH G， MACIACZYK J. Epithelial-to-mesenchymal（-like）transition as a relevant molecular event in malignant gliomas［J］. Cancer Lett， 2013， 331（2）： 131-138.
2	ISER I C， PEREIRA M B， LENZ G， et al. The epithelial-to-mesenchymal transition-like process in glioblastoma： an updated systematic review and in silico investigation［J］. Med Res Rev， 2017， 37（2）： 271-313.
3	DESHPANDE R P， BABU P P. pDok2， caspase 3 dependent glioma cell growth arrest by nitidine chloride［J］. Pharmacol Rep， 2018， 70（1）： 48-54.
4	CUI Y， WU L H， CAO R， et al. Antitumor functions and mechanisms of nitidine xhloride in human cancers［J］. J Cancer， 2020， 11（5）： 1250-1256.
5	KIM H， KIM D， CHOI S A， et al. KDM3A histone demethylase functions as an essential factor for activation of JAK2-STAT3 signaling pathway［J］. Proc Natl Acad Sci U S A， 2018， 115（46）： 11766-11771.
6	刘丽敏，刘华钢. 氯化两面针碱的抗肝癌活性及对DNA拓扑异构酶的影响［J］. 中国药理学通报， 2010， 26（4）： 497-500.
7	程翔宇. 氯化两面针碱对前列腺癌细胞抑制增殖、侵袭与诱发凋亡的研究［D］.济南：山东大学， 2015.
8	CHEN S P， YANG L， FENG J. Nitidine chloride inhibits proliferation and induces apoptosis in ovarian cancer cells by activating the Fas signalling pathway［J］. J PharmPharmacol， 2018， 70（6）： 778-786.
9	袁翠林，娄诤，谢璐迪，等. 氯化两面针碱对人食管癌Eca109细胞抑制作用及机制研究［J］. 中草药， 2019， 50（20）： 4969-4973.
10	ALFONSO J C L， TALKENBERGER K， SEIFERT M， et al. The biology and mathematical modelling of glioma invasion： a review［J］. J R Soc Interface， 2017， 14（136）： 20170490.
11	SONG Y， CHEN Y， LI Y Q， et al. Metformin inhibits TGF-β1-induced epithelial-to-mesenchymal transition-like process and stem-like properties in GBM via AKT/mTOR/ZEB1 pathway［J］. Oncotarget， 2018， 9（6）： 7023-7035.
12	SONG Y， CHEN Y， LI Y Q， et al. Resveratrol suppresses epithelial-mesenchymal transition in GBM by regulating smad-dependent signaling［J］. Biomed Res Int， 2019， 2019： 1321973.
13	YEUNG K T，YANG J. Epithelial mesenchymal transition in tumor metastasis［J］.Mol Oncol，2017，11（1）：28-39.
14	ZHENG X F， CARSTENS J L， KIM J， et al. Epithelial-to-mesenchymal transition is dispensable for metastasis but induces chemoresistance in pancreatic cancer［J］. Nature， 2015， 527（7579）： 525-530.
15	FISCHER K R， DURRANS A， LEE S， et al. Epithelial-to-mesenchymal transition is not required for lung metastasis but contributes to chemoresistance［J］. Nature， 2015， 527（7579）： 472-476.
16	SUN M J， ZHANG N， WANG X L， et al. Hedgehog pathway is involved in nitidine chloride induced inhibition of epithelial-mesenchymal transition and cancer stem cells-like properties in breast cancer cells［J］. Cell Biosci， 2016， 6： 44.
17	CHENG Z X， GUO Y L， YANG Y B， et al. Nitidine chloride suppresses epithelial-to-mesenchymal transition in osteosarcoma cell migration and invasion through Akt/GSK-3β/Snail signaling pathway［J］. Oncol Rep， 2016， 36（2）： 1023-1029.
18	FU X Q， CHOU J Y， LI T， et al. The JAK2/STAT3 pathway is involved in the anti-melanoma effects of atractylenolide I［J］. Exp Dermatol， 2018， 27（2）： 201-204.
19	MA Y， ZHANG H， XIONG C， et al. CD146 mediates an E-cadherin-to-N-cadherin switch during TGF-β signaling-induced epithelial-mesenchymal transition［J］. Cancer Lett， 2018， 430： 201-214.
20	XIONG H， HONG J， DU W， et al. Roles of STAT3 and ZEB1 proteins in E-cadherin down-regulation and human colorectal cancer epithelial-mesenchymal transition［J］. J Biol Chem， 2012， 287（8）： 5819-5832.
21	ZENG Y T， LIU X F， YANG W T， et al. REX1 promotes EMT-induced cell metastasis by activating the JAK2/STAT3-signaling pathway by targeting SOCS1 in cervical cancer［J］. Oncogene， 2019， 38（43）： 6940-6957.
22	CHEN Y， SHAO Z， JIANG E H， et al. CCL21/CCR7 interaction promotes EMT and enhances the stemness of OSCC via a JAK2/STAT3 signaling pathway［J］. J Cell Physiol， 2020， 235（9）： 5995-6009.
23	WANG L L， ZHANG F， CUI J Y， et al. CAFs enhance paclitaxel resistance by inducing EMT through the IL‑6/JAK2/STAT3 pathway［J］. Oncol Rep， 2018， 39（5）： 2081-2090.
24	CHEN J， WANG J Q， LIN L， et al. Inhibition of STAT3 signaling pathway by nitidine chloride suppressed the angiogenesis and growth of human gastric cancer［J］. Mol Cancer Ther， 2012， 11（2）： 277-287.

Group	Migration rate	Invasion rate
Control	53.384±0.993	100.000±0.000
EMT	71.588±1.005^*	150.594±7.209^*
NC（μmol·L^－1）
2.5	19.951±2.086^△	86.002±7.534^△
5.0	18.122±1.505^△	72.584±8.697^△
7.5	12.722±1.528^△	30.461±2.387^△
10.0	13.265±0.617^△	9.419±1.381^△

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[2]	ZHOU Ning, WU Rui, MA Zhenkai, CHEN Weiwei, LI Xuelin, GONG Kaikai, YANG Lijuan, DAI Juanjuan, WU Yan. Effects of down-regulation of ADAR1 expression on proliferation, migration and epithelial-mesenchymal transition of human lung cancer cells [J]. Journal of Jilin University(Medicine Edition), 2020, 46(04): 669-674.
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[5]	DENG Haiyue, YU Fang, JIANG Hong. Regulatory effect of direct renin inhibitor aliskiren on epithelial-mesenchymal transition of renal fibrosis mice [J]. Journal of Jilin University Medicine Edition, 2017, 43(01): 16-20.
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[11]	XU Jun-jie,U Hong-quan,ZHAO Wei,JIN Hong,WEN Na,QI Ling，LIU　Xing-ji. Induction of broccoli polypeptide on apoptosis of C6 glioma cells [J]. Journal of Jilin University Medicine Edition, 2013, 39(1): 8-11.
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[13]	SHANG　Jing,FANG Ning\|TENG Si-ying\|CUI Xiang-yan\|WANG Xin. Expressions of Snail and E-cadherin in laryngeal squamous cell carcinoma tissues and their significances [J]. J4, 2012, 38(2): 315-319.
[14]	ZHANG Shi, ZHANG Yue, BI Xiao-Ying, LI Zhi-Chao. Inhibitory effect of triethyltin chloride on proliferation of \|rat C6 glioma cells in vitro [J]. J4, 2009, 35(4): 595-598.
[15]	CHI Xiu-mei,ZHANG Ji-zhou,KAN Mu-jie,HONG Min. Effect of heroin on gene expression of PCNA in C6 glioma cells [J]. J4, 2008, 34(6): 949-951.

Inhibitory effect of nitidine chloride on epithelial-mesenchymal transition of glioma cells through JAK2/STAT3 signaling pathway

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