|Table of Contents|

Silencing or inhibiting TAK1 expression can improve paclitaxel resistance in ovarian cancer cells through the NF-κB signaling pathway

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2022 04
Page:
576-582
Research Field:
Publishing date:

Info

Title:
Silencing or inhibiting TAK1 expression can improve paclitaxel resistance in ovarian cancer cells through the NF-κB signaling pathway
Author(s):
WANG Qinghui1; LI Bo1; HU Chuancui1; NIE Mingchao1; ZHENG Xiaomei2; JIN Kelong1
1.Department of Obstetrics and Gynecology,Hainan Women and Children's Medical Center,Hainan Haikou 570206，China;2.Department of Obstetrics and Gynecology,the First Affiliated Hospital of Hainan Medical College,Hainan Haikou 570102，China.
Keywords:
ovarian cancer; transforming growth factor-β (TGF-β)-activated kinase 1(TAK1); paclitaxel; NF-κB signaling pathway
PACS:
R737.31
DOI:
10.3969/j.issn.1672-4992.2022.04.004
Abstract:
Objective:To investigate the effect of targeted silencing or inhibiting TAK1 expression of small interfering RNA (siRNA) or transforming growth factor-β (TGF-β)-activated kinase 1 (TAK1) inhibitor 5Z-7-oxozeaenol on taxol resistance of ovarian cancer cells and its mechanism.Methods:RT-PCR and Western blot were used to detect the expression of TAK1 in normal ovarian epithelial cells and ovarian cancer cell lines,and OVCAR3 and SKOV3 cells with higher and lower expression levels of TAK1 mRNA and protein were screened for further study.The expression of TAK1 in OVCAR3 and SKOV3 cells was down regulated by siRNA transfection,and the transfection efficiency was verified by RT-PCR.OVCAR3 or SKOV3 cells were divided into three groups:si-NC+taxol group,si-TAK1+taxol group and si-NC+taxol+5Z-7-oxozeaenol group.CCK-8 method was used to detect the IC50 value of taxol on ovarian cells in each group.Tunel fluorescence staining was used to detect the incidence of apoptosis mediated by taxol on ovarian cells in each group.Phosphorylation levels of NF-κB signaling pathway IκBα (p-IκBα/IκBα) and p65 (p-p65/p65) and expression of apoptotic related protein cleaved caspase-3 were detected by Western blot.SKOV3 cells were used to establish subcutaneous xenograft tumor in nude mice with ovarian cancer,and the mice were divided into four groups:Control group,si-NC+taxol group,si-TAK1+taxol group and si-NC+taxol+5Z-7-oxozeaenol group.Tumor growth were monitored,and the expression of TAK1 in tumor tissues was detected by immunohistochemistry.Cell apoptosis rate,phosphorylation of NF-κB signaling pathway IκBα and p65,and cleaved caspase-3 protein expression were detected by Tunel fluorescence staining and Western blot.Results:Compared with normal ovarian epithelial cells,the expression levels of TAK1 mRNA and protein in ovarian cancer cell lines were significantly increased (P＜0.05),and the highest expression level was in OVCAR3 cells,while the lowest expression level was in SKOV3 cells.The expression of TAK1 in OVCAR3 and SKOV3 cells was down regulated by siRNA transfection.Compared with si-NC+taxol group,the IC50 value of paclitaxel in si-TAK1+taxol group and si-NC+taxol+5Z-7-oxozeaenol group was significantly decreased (P＜0.05),and the apoptosis rate was significantly increased (P＜0.05).The phosphorylation levels of IκBα and p65 were significantly decreased (P＜0.05),while the protein expression level of cleaved caspase-3 was significantly increased (P＜0.05).Compared with control group,the tumor weight and growth rate of si-NC+taxol group,si-TAK1+taxol group and si-NC+taxol+5Z-7-oxozeaenol group were significantly decreased (P＜0.05),and the protein expression of TAK1 in tumor tissue was significantly decreased (P＜0.05).The phosphorylation level of IκBα and p65 was also significantly decreased (P＜0.05),while the apoptosis rate and cleaved caspase-3 protein expression level were significantly increased (P＜0.05).Compared with the si-NC+taxol group,the change trend of the above detection indicators was more significant in the si-TAK1+taxol group and si-NC+taxol+5Z-7-oxozeaenol group (P＜0.05).Conclusion:In vivo and in vitro experiments showed that TAK1 is an important molecule for the development of taxol resistance in ovarian cancer,and silenced or inhibited TAK1 expression may promote taxol-induced apoptosis by inhibiting the activation level of NF-κB signaling pathway,so as to improve the sensitivity of tumors to taxol chemotherapy.

References:

[1] MATULONIS UA.Ovarian cancer［J］.Hematol Oncol Clin North Am,2018,32(6):13-19.
[2] WANG GS,WANG Y.Research progress on the relationship between the microenvironment of ovarian cancer and the occurrence of chemotherapy resistance［J］.Cancer,2020,40(1):68-75.
[3] ZHAO MD,LI JQ,CHEN FY,et al.Co-delivery of curcumin and paclitaxel by "core-shell" targeting amphiphilic copolymer to reverse resistance in the treatment of ovarian cancer［J］.Int J Nanomedicine,2019,14:9453-9467.
[4] MORIOKA S,BROGLIE P,OMORI E,et al.TAK1 kinase switches cell fate from apoptosis to necrosis following TNF stimulation［J］.J Cell Biol,2014,204(4):607-623.
[5] SAKURAI H.Targeting of TAK1 in inflammatory disorders and cancer［J］.Trends Pharmacol Sci,2012,33(10):522-530.
[6] BO L,CUI H,FANG Z,et al.Inactivation of transforming growth factor-β-activated kinase 1 promotes taxol efficacy in ovarian cancer cells［J］.Biomed Pharmacother,2016,84(015):917-924.
[7] XUE T,WANG LM,JIAO JW,et al.siRNA-mediated RRM2 gene silencing in the treatment of human ovarian cancer transplanted tumors in nude mice［J］.Journal of Shandong University (Health Sciences),2019,57(10):74-79.
[8] YIN S,ZENG C,HARI M,et al.Paclitaxel resistance by random mutagenesis of α-tubulin［J］.Cytoskeleton (Hoboken),2013,70(12):849-862.
[9] PU J,SHEN J,ZHONG Z,et al.KANK1 regulates paclitaxel resistance in lung adenocarcinoma A549 cells［J］.Artif Cells Nanomed Biotechnol,2020,48(1):639-647.
[10] WILLIAMS LM,GILMORE TD.Looking down on NF-κB［J］.Mol Cell Biol,2020,40(15):e00104-e00120.
[11] PERKINS ND.The diverse and complex roles of NF-κB subunits in cancer［J］.Nat Rev Cancer,2012,12(2):121-132.
[12] BEN-NERIAH Y,KARIN M.Inflammation meets cancer,with NF-κB as the matchmaker［J］.Nat Immunol,2011,12(8):715-723.
[13] DENG Y,LU J,LI W,et al.Reciprocal inhibition of YAP/TAZ and NF-κB regulates osteoarthritic cartilage degradation［J］.Nat Commun,2018,9(1):e4564.
[14] SUN XL,WANG ZL,WU Q,et al.LncRNA RMST activates TAK1-mediated NF-κB signaling and promotes activation of microglial cells via competitively binding with hnRNPK［J］.IUBMB Life,2019,71(11):1785-1793.

Memo

Memo:
海南省基础与应用基础研究计划（自然科学领域）高层次人才项目（编号：2019RC391）
Last Update: 1900-01-01