Volume 28, Issue 1 (Avicenna Journal of Clinical Medicine-Spring 2021)                   Avicenna J Clin Med 2021, 28(1): 49-58 | Back to browse issues page

XML Persian Abstract Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Haghighi N, Doosti A, Kiani J. Therapeutic Efficacy Analysis of lncRNA NEAT1 Gene Knockout and Apoptosis Induction in Prostate Cancer Cell Line Using CRISPR/Cas9. Avicenna J Clin Med 2021; 28 (1) :49-58
URL: http://sjh.umsha.ac.ir/article-1-2204-en.html
Therapeutic Efficacy Analysis of lncRNA NEAT1 Gene Knockout and Apoptosis Induction in Prostate Cancer Cell Line Using CRISPR/Cas9
Nastaran Haghighi1 , Abbas Doosti 2, Jafar Kiani3
1- PhD Student in Molecular Genetics, Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
2- Associate Professor, Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran , abbasdoosti@yahoo.com
3- Assistant Professor, Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
Abstract:   (2118 Views)
Background and Objective: Long non-coding ribonucleic acid (lncRNA) has been identified as an important gene regulator and prognostic marker in various cancers. The present study aimed to investigate the effects of Nuclear Paraspeckle Assembly Transcript1 (NEAT1) gene knockout using Clustered Regularly Interspaced Short Palindromic Repeats-associated Protein 9 (CRISPR/Cas9) in PC-3 cell line.
Materials and Methods: In this experimental study, recombinant plasmids carrying single guide RNA in the CRISPR system were designed and constructed. The prostate cancer PC3 cell lines were transfected by recombinant vectors, and apoptotic gene expression was evaluated using the quantitative polymerase chain reaction technique. Furthermore, annexin, MTT, and wound healing assays were applied for apoptosis, cell proliferation, and cell migration activities respectively. 
Results: Knockout of NEAT1 gene using CRISPR/Cas9 technique caused significant changes in the expression of apoptosis-related genes in a way that the expression of P21, BCL2, and BIRC5 genes decreased, while the expression of P53 and BAX increased. In addition, in the treatment group, the rate of cell proliferation and migration was reduced compared to those in the control group. Moreover, an increase was observed in the apoptosis rate in genetically modified cells compared to the control group.
Conclusion: As an oncogene, NEAT1 plays an important role in tumorigenesis and its knockout reduces cell survival and migration and increases the apoptosis of cancer cells.
 
Keywords: CRISPR-Associated Protein 9, NEAT1 Long Non-coding RNA, Prostatic Neoplasms, PC-3 Cells
Full-Text [PDF 1293 kb]   (1401 Downloads)    
Type of Study: Original | Subject: Molecular Medicine & Biotechnology
Extended Abstract [HTML 29 KB]  (84 Download)
References
1. Obertova Z, Brown C, Holmes M, Lawrenson R. Prostate cancer incidence and mortality in rural men–a systematic review of the literature. Rural Remote Health. 2012;12(2):2039-51. PMID: 22616627
2. Rebbeck TR. Prostate cancer genetics: variation by race, ethnicity, and geography. Semin Radiat Oncol. 2017;27(1):3-10. PMID: 27986209 DOI: 10.1016/j.semradonc.2016.08.002
3. Dunn MW. Prostate cancer screening. Semin Oncol Nurs. 2017;33(2):156-64. PMID: 28343840 DOI: 10.1016/j.soncn.2017.02.003
4. Xing Y, Yao R, Zhang Y, Guo C, Jiang S, Xu G, et al. SLERT regulates DDX21 rings associated with Pol I transcription. Cell. 2017;169(4):664-78. PMID: 28475895 DOI: 10.1016/j.cell.2017.04.011
5. Ning L, Li Z, Wei D, Chen H, Yang C. LncRNA, NEAT1 is a prognosis biomarker and regulates cancer progression via epithelial-mesenchymal transition in clear cell renal cell carcinoma. Cancer Biomark. 2017;19(1):75-83. PMID: 28269753 DOI: 10.3233/CBM-160376
6. Li P, Huang R, Huang T, Cheng S, Chen Y, Wang Z. Long non-coding RNA NEAT1 promotes proliferation, migration and invasion of human osteosarcoma cells. Int J Med Sci. 2018;15(11):1227-34. PMID: 30123061 DOI: 10.7150/ijms.25662
7. Fang J, Qiao F, Tu J, Xu J, Ding F, Liu Y, et al. High expression of long non-coding RNA NEAT1 indicates poor prognosis of human cancer. Oncotarget. 2017;8(28):45918-27. PMID: 28507281 DOI: 10.18632/oncotarget.17439
8. Fu JW, Kong Y, Sun X. Long noncoding RNA NEAT1 is an unfavorable prognostic factor and regulates migration and invasion in gastric cancer. J Cancer Res Clin Oncol. 2016;142(7):1571-9. PMID: 27095450 DOI: 10.1007/s00432-016-2152-1
9. Wang Z, Zou Q, Song M, Chen J. NEAT1 promotes cell proliferation and invasion in hepatocellular carcinoma by negative regulating miR-613 expression. Biomed Pharmacother. 2017;94:612-18. PMID: 28783584 DOI: 10.1016/j.biopha.2017.07.111
10. Xia TF, Chen J, Wu K, Zhang J, Yan Q. Long noncoding RNA NEAT1 promotes the growth of gastric cancer cells by regulating miR-497-5p/PIK3R1 axis. Eur Rev Med Pharmacol Sci. 2019;23(16):6914-26. PMID: 31486491 DOI: 10.26355/eurrev_201908_18731
11. Wei C, Wang F, Liu W, Zhao W, Yang Y, Li K, et al. CRISPR/Cas9 targeting of the androgen receptor suppresses the growth of LNCaP human prostate cancer cells. Mol Med Rep. 2018;17(2):2901-6. PMID: 29257308 DOI: 10.3892/mmr.2017.8257
12. Li X, Wang X, Song W, Xu H, Huang R, Wang Y, et al. Oncogenic properties of NEAT1 in prostate cancer cells depend on the CDC5L–AGRN transcriptional regulation circuit. Cancer Res. 2018;78(15):4138-49. PMID: 29871935 DOI: 10.1158/0008-5472.CAN-18-0688
13. Chen B, Deng S, Ge T, Ye M, Yu J, Lin S, et al. Live cell imaging and proteomic profiling of endogenous NEAT1 lncRNA byCRISPR/Cas9-mediated knock-in. Protein Cell. 2020;11(9):641-60. PMID: 32458346 DOI: 10.1007/s13238-020-00706-w
14. Esposito R, Bosch N, Lanzós A, Polidori T, Pulido-Quetglas C, Johnson R. Hacking the cancer genome: profiling therapeutically actionable long non-coding RNAs using CRISPR-Cas9 screening. Cancer Cell. 2019;35(4):545-57. PMID: 30827888 DOI: 10.1016/j.ccell.2019.01.019
15. Tai Sh, Sun Y, Squires J, Zhang H, Oh W, Liang C, et al. PC3 is a cell line characteristic of prostatic small cell carcinoma. Prostate. 2011;71(15):1668-79. PMID: 21432867 DOI: 10.1002/pros.21383
16. Ghasemi-Dehkordi P, Doosti A, Jami MS. The functions of azurin of Pseudomonas aeruginosa and human mammaglobin-A on proapoptotic and cell cycle regulatory genes expression in the MCF-7 breast cancer cell line. Saudi J Biol Sci. 2020;27(9):2308-17. PMID: 32884412 DOI: 10.1016/j.sjbs.2020.04.007
17. Abedi S, Doosti A, Jami MS. Evaluation of the preventive and therapeutic effects of a recombinant vector co‐expressing prostate‐specific stem cell antigen (PSCA) and Clostridium perfringens enterotoxin (CPE) on prostate cancer in rats. Biotechnol Prog. 2020;36(2):e2906. PMID: 31513734 DOI: 10.1002/btpr.2906
18. Wang X, Decker CC, Zechner L, Krstin S, Wink M. In vitro wound healing of tumor cells: inhibition of cell migration by selected cytotoxic alkaloids. BMC Pharmacol Toxicol. 2019;20(1):4-12. PMID: 30626448 DOI: 10.1186/s40360-018-0284-4
19. Ma Y, Liu L, Yan F, Wei W, Deng J, Sun J. Enhanced expression of long non-coding RNA NEAT1 is associated with the progression of gastric adenocarcinomas. World J Surg Oncol. 2016;14(1):41-6. PMID: 26911892 DOI: 10.1186/s12957-016-0799-3
20. Yu DJ, Guo CX, Qian J, Li J, Zhu C, Jin X, et al. The long non-coding RNA NEAT1 promotes gastric cancer cell proliferation and invasion by regulating miR-103a/STAMBPL1 axis. Technol Cancer Res Treat. 2020;19:1-8. PMID: 33111649 DOI: 10.1177/1533033820964081
21. Shamloo B, Usluer S. p21 in cancer research. Cancers (Basel). 2019;11(8):1178-97. PMID: 31416295 DOI: 10.3390/cancers11081178
22. Wu D, Li H, Wang J, Li H, Xiao Q, Zhao X, et al. LncRNA NEAT1 promotes gastric cancer progression via miR-1294/AKT1 axis. Open Med. 2020;15(1):1028-38. PMID: 33336058 DOI: 10.1515/med-2020-0218
23. Campbell KJ, Tait SW. Targeting BCL-2 regulated apoptosis in cancer. Open Biol. 2018;8(5):180002. PMID: 29769323 DOI: 10.1098/rsob.180002
24. Li Y, Pan J, Gou M. The anti-proliferation, cycle arrest and apoptotic inducing activity of peperomin E on prostate cancer PC-3 cell line. Molecules. 2019;24(8):1472. PMID: 30991627 DOI: 10.3390/molecules24081472
25. Li F, Aljahdali I, Ling X. Cancer therapeutics using survivin BIRC5 as a target: what can we do after over two decades of study? J Exp Clin Cancer Res. 2019;38(1):368. PMID: 31439015 DOI: 10.1186/s13046-019-1362-1
26. Shang X, Liu G, Zhang Y, Tang P, Zhang H, Jiang H, et al. Downregulation of BIRC5 inhibits the migration and invasion of esophageal cancer cells by interacting with the PI3K/Akt signaling pathway. Oncol Lett. 2018;16(3):3373-9. PMID: 30127937 DOI: 10.3892/ol.2018.8986
Add your comments about this article : Your username or Email:
CAPTCHA
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2024 CC BY-NC 4.0 | Avicenna Journal of Clinical Medicine

Designed & Developed by : Yektaweb