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Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma
Published on: April 20, 2018
TCAB1: a potential target for diagnosis and therapy of head and neck carcinomas
Chong-kui Sun, Xiao-bo Luo, Ya-ping Gou
1State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, China. xiaoly@scu.edu.cn.
Background:
WRAP53, including α, β and γ isoforms, plays an important role not only in the stability of p53 mRNA, but also in the assembly and trafficking of the telomerase holoenzyme. It has been considered an oncogene and is thought to promote the survival of cancer cells. The aim of this study was to detect the role of TCAB1 (except WRAP53α) in the occurrence and development of head and neck carcinomas.
Methods:
Immunohistochemistry was used to detect the TCAB1 expression in clinical specimen sections and performed western blotting to check the TCAB1 expression levels in cell lines. TCAB1 was depleted using shRNA lentivirus and the knockdown efficiency was assessed using q-PCR and Western blotting. We performed CCK-8 assays and flow cytometry to check the cell proliferation potential and used the trans-well assay to test the invasion ability in vitro. Xenografts were used to detect the tumor formation potential in vivo. Moreover, we performed cDNA microarray to investigate the candidate factors involved in this process.
Results:
We observed a notable overexpression of TCAB1 in head and neck carcinoma clinical specimens as well as in carcinoma cell lines. Knockdown of TCAB1 decreased the cellular proliferation potential and invasion ability in vitro. cDNA microarray analysis suggested the possible involvement of several pathways and factors associated with tumorigenesis and carcinoma development in the TCAB1-mediated regulation of cancers. Furthermore, the xenograft assay confirmed that the depletion of TCAB1 would inhibit tumor formation in nude mice. The immunohistochemistry results of the mice tumor tissue sections revealed that the cells in shTCAB1 xenografts showed decreased proliferation potential and increased apoptotic trend, meanwhile, the angiogenesis was inhibited in the smaller tumors form shTCAB1 cells.
Conclusions:
Our study demonstrated that depletion of TCAB1 decreased cellular proliferation and invasion potential both in vitro and in vivo. The data indicated that TCAB1 might facilitate the occurrence and development of head and neck carcinomas. In future, TCAB1 might be useful as a prognostic biomarker or a potential target for the diagnosis and therapy of head and neck carcinomas.
Insights
TCAB1 is overexpressed in head and neck cancers and promotes tumor growth and spread. Depleting TCAB1 inhibits cancer cell proliferation and invasion, suggesting it as a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
Background:
- WRAP53 (including α, β, γ isoforms) is implicated in p53 mRNA stability and telomerase assembly.
- WRAP53 is considered an oncogene promoting cancer cell survival.
Purpose of the Study:
- To investigate the role of TCAB1 (excluding WRAP53α) in head and neck carcinomas.
- To determine TCAB1's involvement in cancer occurrence and development.
Main Methods:
- Immunohistochemistry and Western blotting for TCAB1 expression analysis.
- shRNA-mediated TCAB1 depletion, assessed by q-PCR and Western blotting.
- In vitro assays (CCK-8, flow cytometry, trans-well) for proliferation and invasion.
- In vivo xenograft models for tumor formation and angiogenesis assessment.
- cDNA microarray for pathway analysis.
Main Results:
- TCAB1 is significantly overexpressed in head and neck carcinoma specimens and cell lines.
- TCAB1 depletion reduced cell proliferation and invasion in vitro.
- TCAB1 knockdown inhibited tumor formation, proliferation, and angiogenesis, while increasing apoptosis in vivo.
- Microarray analysis identified pathways linked to TCAB1-mediated tumorigenesis.
Conclusions:
- TCAB1 depletion inhibits head and neck carcinoma cell proliferation and invasion.
- TCAB1 plays a facilitating role in the occurrence and development of head and neck carcinomas.
- TCAB1 may serve as a prognostic biomarker and therapeutic target for head and neck carcinomas.
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