Establishment of a Livin-gene silencing system and its effect on the sensitivity of Cal 27 with regard to 5-fluorouracil
DOI: 10.54647/cm32648 82 Downloads 4487 Views
Author(s)
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
References
[ 1 ] | Bray, F. et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians 68, 394-424, doi:10.3322/caac.21492 (2018). |
[ 2 ] | Chen, W. et al. Cancer statistics in China, 2015. CA: a cancer journal for clinicians 66, 115-132, doi:10.3322/caac.21338 (2016). |
[ 3 ] | Ferlay, J. et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136, E359-386, doi:10.1002/ijc.29210 (2015). |
[ 4 ] | Shield, K. D. et al. The global incidence of lip, oral cavity, and pharyngeal cancers by subsite in 2012. CA: a cancer journal for clinicians 67, 51-64, doi:10.3322/caac.21384 (2017). |
[ 5 ] | Azharuddin, M. et al. Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids. Scientific Reports 9, 20066, doi:10.1038/s41598-019-56273-6 (2019). |
[ 6 ] | Vasan, N., Baselga, J. & Hyman, D. M. A view on drug resistance in cancer. Nature 575, 299-309, doi:10.1038/s41586-019-1730-1 (2019). |
[ 7 ] | Bell, C. C. & Gilan, O. Principles and mechanisms of non-genetic resistance in cancer. British Journal of Cancer, doi:10.1038/s41416-019-0648-6 (2019). |
[ 8 ] | Altieri, B. et al. Livin/BIRC7 expression as malignancy marker in adrenocortical tumors. Oncotarget 8, 9323-9338, doi:10.18632/oncotarget.14067 (2017). |
[ 9 ] | Wang, H. et al. Single-chain antibody-delivered Livin siRNA inhibits human malignant melanoma growth in vitro and in vivo. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 39, 1010428317701645, doi:10.1177/1010428317701645 (2017). |
[ 10 ] | Su, Q. B. et al. Livin serves as a prognostic marker for mid-distal rectal cancer and a target of mid-distal rectal cancer treatment. Oncology letters 14, 7759-7766, doi:10.3892/ol.2017.7230 (2017). |
[ 11 ] | Wang, Z. et al. Silencing Livin induces apoptotic and autophagic cell death, increasing chemotherapeutic sensitivity to cisplatin of renal carcinoma cells. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 37, 15133-15143, doi:10.1007/s13277-016-5395-1 (2016). |
[ 12 ] | Zhuang, L. et al. Inhibition of livin expression suppresses cell proliferation and enhances chemosensitivity to cisplatin in human lung adenocarcinoma cells. Molecular medicine reports 12, 547-552, doi:10.3892/mmr.2015.3372 (2015). |
[ 13 ] | Elmekkawy, B. K., Shoaib, R. M. S., Seleem, A. K., Shalaan, D. & Saad, E. A. Livin/BIRC7 gene expression as a possible diagnostic biomarker for endometrial hyperplasia and carcinoma. Journal, genetic engineering & biotechnology 19, 141, doi:10.1186/s43141-021-00244-w (2021). |
[ 14 ] | Fung, S., Knoefel, W. T. & Krieg, A. Clinicopathological and Prognostic Significance of Inhibitor of Apoptosis Protein (IAP) Family Members in Lung Cancer: A Meta-Analysis. Cancers 13, doi:10.3390/cancers13164098 (2021). |
[ 15 ] | Ge, Y., Liu, B.-l., Cui, J.-p. & Li, S.-q. Livin promotes colon cancer progression by regulation of H2A.XY39ph via JMJD6. Life Sciences 234, 116788, doi:https://doi.org/10.1016/j.lfs.2019.116788 (2019). |
[ 16 ] | Liu, S. et al. Silencing Livin improved the sensitivity of colon cancer cells to 5-fluorouracil by regulating crosstalk between apoptosis and autophagy. Oncology letters 15, 7707-7715, doi:10.3892/ol.2018.8282 (2018). |
[ 17 ] | Uegaki, T. et al. Inhibitor of apoptosis proteins (IAPs) may be effective therapeutic targets for treating endometriosis. Human reproduction (Oxford, England) 30, 149-158, doi:10.1093/humrep/deu288 (2015). |
[ 18 ] | Xue, J. M. et al. Livin in synergy with Ras induces and sustains corticosteroid resistance in the airway mucosa. International journal of biological sciences 17, 2089-2098, doi:10.7150/ijbs.58427 (2021). |
[ 19 ] | Jin, J. et al. Bioinformatics analysis of aberrantly expressed exosomal lncRNAs in oral squamous cell carcinoma (CAL-27 vs. oral epithelial) cells. Oncology letters 20, 2378-2386, doi:10.3892/ol.2020.11764 (2020). |
[ 20 ] | Qin, X. et al. Mechanism and significance of apoptosis of the immortalized human oral mucosal epithelial cells established by Lentivirus-mediated hTERT. Mol Biol Rep 47, 5469-5475, doi:10.1007/s11033-020-05637-7 (2020). |
[ 21 ] | Sun, G. et al. Comparison of Periodontal Ligament Cell Lines with Adenovirus- and Lentivirus-Mediated Human Telomerase Reverse Transcription Expression. Human gene therapy methods 30, 53-59, doi:10.1089/hgtb.2018.184 (2019). |
[ 22 ] | Hrdinka, M. & Yabal, M. Inhibitor of apoptosis proteins in human health and disease. Genes and immunity 20, 641-650, doi:10.1038/s41435-019-0078-8 (2019). |
[ 23 ] | Nakajima, Y. I. & Kuranaga, E. Caspase-dependent non-apoptotic processes in development. Cell death and differentiation 24, 1422-1430, doi:10.1038/cdd.2017.36 (2017). |
[ 24 ] | Li, P. et al. Caspase-9: structure, mechanisms and clinical application. Oncotarget 8, 23996-24008, doi:10.18632/oncotarget.15098 (2017). |
[ 25 ] | Green, D. R. & Llambi, F. Cell Death Signaling. Cold Spring Harbor perspectives in biology 7, doi:10.1101/cshperspect.a006080 (2015). |
[ 26 ] | Kasof, G. M. & Gomes, B. C. Livin, a novel inhibitor of apoptosis protein family member. J Biol Chem 276, 3238-3246, doi:10.1074/jbc.M003670200 (2001). |
[ 27 ] | Jin, F. et al. Difference in the Inhibitory Effect of Temozolomide on TJ905 Glioma Cells and Stem Cells. Frontiers in neurology 8, 474, doi:10.3389/fneur.2017.00474 (2017). |
[ 28 ] | Yang, D. et al. Therapeutic potential of siRNA-mediated combined knockdown of the IAP genes (Livin, XIAP, and Survivin) on human bladder cancer T24 cells. Acta biochimica et biophysica Sinica 42, 137-144, doi:10.1093/abbs/gmp118 (2010). |
[ 29 ] | Li, G. et al. Effect of temozolomide on livin and caspase-3 in U251 glioma stem cells. Experimental and therapeutic medicine 9, 744-750, doi:10.3892/etm.2014.2144 (2015). |
[ 30 ] | Liang, Y. et al. miR-198-induced upregulation of Livin may be associated with the prognosis and contribute to the oncogenesis of lung adenocarcinoma. Oncol Rep 38, 2096-2104, doi:10.3892/or.2017.5866 (2017). |