Establishment of a Livin-gene silencing system and its effect on the sensitivity of Cal 27 with regard to 5-fluorouracil

Zixiao Huang, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China;Xiangyang Stomatological Hospital; Affiliated Stomatological Hospital of Hubei University of Arts and Science, Xiangyang, 441000, P. R. China

Ziwei Cui, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China

Ruoshan Qin, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China

Xuanxuan Yao, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China

Hongli Zhou, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China

Ru Guo, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China

Xiaodong Qin, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, 730000, P. R. China

Xiangyi He, School of Dentistry, Lanzhou University, Lanzhou, 730000, P. R. China;Key Laboratory of Functional Genomic and Molecular Diagnosis of Gansu Province, Lanzhou, 730030, P. R. China

Abstract
Objective: This study aims at downregulating the expression of Livin through shRNA and exploring the sensitivity of Cal 27 to 5-fluorouracil (5-FU), so as to provide some help in resolving the chemotherapy resistance of oral cancer.
Methods: The Livin-targeting shRNA sequence was designed to construct a lentivirus and infect Cal 27 to obtain a stable Livin-silencing cell strain. After being treated with 20 μM, 40 μM and 60 μM of 5-fluorouracil for 24 h, cell apoptosis and cell viability were measured by flow cytometry and MTT assay, respectively.
Results: A shRNA-based lentivirus targeting Livin was successfully designed and constructed. After the silencing of Livin, the sensitivity and cell apoptosis of Cal 27 to 5-FU were dramatically elevated, and cell viability was significantly reduced (P<0.05). Moreover, the inhibition of Caspase 3 was observed.
Conclusion: The expression level of Livin was downregulated in Cal 27 after shRNA transfection, which increased the sensitivity of Cal to 5-FU. The underlying mechanism is the blockage of the activation of Caspase 3. Therefore, Livin may serve as a promising target for oral cancer treatment.

Keywords
Mouth Neoplasms; Inhibitor of Apoptosis Proteins; Drug Therapy; Caspase 3; Fluorouracil; Gene Silencing

Cite this paper
Zixiao Huang, Ziwei Cui, Ruoshan Qin, Xuanxuan Yao, Hongli Zhou, Ru Guo, Xiaodong Qin, Xiangyi He, Establishment of a Livin-gene silencing system and its effect on the sensitivity of Cal 27 with regard to 5-fluorouracil , SCIREA Journal of Clinical Medicine. Volume 6, Issue 6, December 2021 | PP. 474-487. 10.54647/cm32648

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