Long non-coding RNA UCA1 Knockdown Assisted by CRISPR/Cas9 in Female Cancer Cell Lines Increases Mir-143 Tumor-Suppressor
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Abstract
Abstract
Background: The lncRNAs has been linked to several malignancies, including breast cancer. Our objective was to investigate the impact of urothelial carcinoma associated 1 (UCA1) on cellular growth and death by a CRISPR/Cas9 knockdown technique.
Methods: In 2020, the CHOPCHOP program was utilized to design two sgRNAs targeting the UCA gene. sgRNA1 and sgRNA2 were inserted into two different CRISPR plasmids to produce two recombinant plasmids. These recombinant plasmids were simultaneously transfected into MCF-7 and MDA-MB 231 carcinoma of the breast cells. Proliferation and apoptosis were compared using the MTT test, CCK-8 assay, and flow cytometry evaluation. RNA-hybrid software, quantitative reverse transcription PCR, and luciferase assays were utilized to confirm the relationship between UCA1 and miR-143.
Results: Proliferated cells were less active in MTT and CCK-8 tests and fellow cytometry analysis. The PX459-sgRNA1,2 group had elevated levels of the cancer biomarker Caspase-3 gene expression (P<0.001). When WT-UCA1 and miR-143 were co-transfected, the luciferase activity was drastically decreased.
Conclusion: One very effective method of regulating cellular proliferation in vitro is the deletion of UCA1, which CRISPR/Cas9 accomplishes.
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15. Nekoei S, Hafshejani TT, Doosti A, et al (2015). Molecular detection of bovine leukemia virus in peripheral blood of Ira-nian cattle, camel and sheep. Pol J Vet Sci, 18(4):703-7.
16. Safarpour-Dehkordi M, Doosti A, Jami MS (2020). Impacts of the Staphylococcal Enterotoxin H on the Apoptosis and lncRNAs in PC3 and ACHN. Mol Gen Microbiol Virol, 35: 180-188.
17. Safarpour-Dehkordi M, Doosti A, Jami MS (2020). Integrative analysis of lncRNAs in kidney cancer to discover a new lncRNA (LINC00847) as a therapeutic target for staphylococcal enterotoxin tst gene. Cell J, 22(Suppl 1):101-109.
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19. Miao Y, Fan R, Chen L, et al (2016). Clinical significance of long non-coding RNA MALAT1 expression in tissue and serum of breast cancer. Ann Clin Lab Sci, 46 (4): 418-424.
20. Soudyab M, Iranpour M, Ghafouri-Fard S (2016). The role of long non-coding RNAs in breast cancer. Arch Iran Med, 19 (7): 508-17.
21. Dhanasekaran R, Deutzmann A, Mahauad-Fernandez, et al (2022). The MYC onco-gene—the grand orchestrator of cancer growth and immune evasion. Nat Rev Clin Oncol, 19 (1): 23-36.
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23. Xu L, Zhu H, Gao F, et al (2019). Upregula-tion of the long non-coding RNA CBR3-AS1 predicts tumor prognosis and con-tributes to breast cancer progression. Gene, 721S:100014..
24. Wang H, Guan Z, He K, et al (2017). LncRNA UCA1 in anti-cancer drug re-sistance. Oncotarget, 8 (38): 64638- 64650.
25. Wang F, Li X, Xie X, et al (2008). UCA1, a non-protein-coding RNA up-regulated in bladder carcinoma and embryo, influenc-ing cell growth and promoting invasion. FEBS Lett, 582 (13): 1919-1927.
26. Xiao C, Wu CH, Hu HZ (2016). LncRNA UCA1 promotes epithelial-mesenchymal transition (EMT) of breast cancer cells via enhancing Wnt/beta-catenin signaling pathway. Eur Rev Med Pharmacol Sci, 20 (13): 2819-2824.
27. Li Y, Zeng Q, Qiu J, et al (2019). Long non-coding RNA UCA1 promotes breast cancer by upregulating PTP1B expression via inhibiting miR-206. Cancer Cell Int, 19:275.
28. Xin H, Liu N, Xu X, et al (2019). Knock-down of lncRNA‐UCA1 inhibits cell via-bility and migration of human glioma cells by miR‐193a‐mediated downregula-tion of CDK6. J Cell Biochem, 120 (9): 15157-15169.
29. Liu C, Jin J, Shi J, et al (2019). Long noncoding RNA UCA1 as a novel bi-omarker of lymph node metastasis and prognosis in human cancer: a meta-analysis. Biosci Rep, 39 (4): BSR20180995
30. Piri-Gharaghie T (2021). Polycystic ovary syndrome and genetic factors influencing its development: A review article. Pers M J, 6 (23): 25-29.
2. Zhuo Li 1, Dehai Yu 2, Haijun Li, et al (2019). Long non coding RNA UCA1 confers tamoxifen resistance in breast cancer endocrinotherapy through regula-tion of the EZH2/p21 axis and the PI3K/AKT signaling pathway. Int J Oncol, 54 (3): 1033-1042.
3. Choudhry H (2021). UCA1 Overexpression Promotes Hypoxic Breast Cancer Cell Proliferation and Inhibits Apoptosis via HIF-1α Activation. J Oncol, 2021: 5512156.
4. Yang G, Shi R, Zhang Q (2020). Hypoxia and oxygen-sensing signaling in gene regulation and cancer progression. Int J Mol Sci, 21 (21): 8162.
5. Moezi P, Kargar M, Doosti A, et al (2019). Multiplex touchdown PCR assay to en-hance specificity and sensitivity for con-current detection of four foodborne pathogens in raw milk. J Appl Microbiol, 127 (1): 262-273.
6. Wang J, Zhang X, Chen W, et al (2020). Regulatory roles of long noncoding RNAs implicated in cancer hallmarks. Int J Cancer, 146 (4): 906-916.
7. Hosseini NF, Manoochehri H, Khoei SG, et al (2021). The functional role of long non-coding RNA UCA1 in human mul-tiple cancers: a review study. Curr Mol Med, 21 (2): 96-110.
8. Neve B, Jonckheere N, Vincent A, et al (2018). Epigenetic regulation by lncRNAs: an overview focused on UCA1 in colo-rectal cancer. Cancers (Basel), 10 (11): 440.
9. Doosti A, Amini-Bavil-Olyaee S, Tajbakhsh E, et al (2009). Prevalence of viral hepati-tis and molecular analysis of HBV among voluntary blood donors in west Iran. New Microbiol, 32 (2): 193-8.
10. Wang ZQ, Cai Q, Hu L, et al (2017). Long noncoding RNA UCA1 induced by SP1 promotes cell proliferation via recruiting EZH2 and activating AKT pathway in gastric cancer. Cell Death Dis, 8 (6): e2839.
11. Kargar M, Rashidi A, Doosti A, et al (2012). Prevalence of Coxiella burnetii in bovine bulk milk samples in southern Iran. Comp Clin Pathol, 22 (3): 331-334.
12. Zhou Y, Meng X, Chen S, et al (2018). IMP1 regulates UCA1-mediated cell inva-sion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p. Breast Cancer Res, 20: 32.
13. Latgé G, Poulet C, Bours V, et al (2018). Natural antisense transcripts: molecular mechanisms and implications in breast cancers. Int J Mol Sci, 19 (1): 123.
14. Huang J, Zhou N, Watabe K, et al (2014). Long non-coding RNA UCA1 promotes breast tumor growth by suppression of p27 (Kip1). Cell Death Dis, 5 (1): e1008.
15. Nekoei S, Hafshejani TT, Doosti A, et al (2015). Molecular detection of bovine leukemia virus in peripheral blood of Ira-nian cattle, camel and sheep. Pol J Vet Sci, 18(4):703-7.
16. Safarpour-Dehkordi M, Doosti A, Jami MS (2020). Impacts of the Staphylococcal Enterotoxin H on the Apoptosis and lncRNAs in PC3 and ACHN. Mol Gen Microbiol Virol, 35: 180-188.
17. Safarpour-Dehkordi M, Doosti A, Jami MS (2020). Integrative analysis of lncRNAs in kidney cancer to discover a new lncRNA (LINC00847) as a therapeutic target for staphylococcal enterotoxin tst gene. Cell J, 22(Suppl 1):101-109.
18. Liu R, Liao J, Yang M, et al (2012). The clus-ter of miR-143 and miR-145 affects the risk for esophageal squamous cell carci-noma through co-regulating FAScin homolog 1. PLoS One, 7 (3): e33987.
19. Miao Y, Fan R, Chen L, et al (2016). Clinical significance of long non-coding RNA MALAT1 expression in tissue and serum of breast cancer. Ann Clin Lab Sci, 46 (4): 418-424.
20. Soudyab M, Iranpour M, Ghafouri-Fard S (2016). The role of long non-coding RNAs in breast cancer. Arch Iran Med, 19 (7): 508-17.
21. Dhanasekaran R, Deutzmann A, Mahauad-Fernandez, et al (2022). The MYC onco-gene—the grand orchestrator of cancer growth and immune evasion. Nat Rev Clin Oncol, 19 (1): 23-36.
22. Piri-Gharaghie T, Doosti A, Mirzaei SA (2023). Novel adjuvant nano-vaccine in-duced immune response against Aci-netobacter baumannii. AMB Express, 13 (1): 31.
23. Xu L, Zhu H, Gao F, et al (2019). Upregula-tion of the long non-coding RNA CBR3-AS1 predicts tumor prognosis and con-tributes to breast cancer progression. Gene, 721S:100014..
24. Wang H, Guan Z, He K, et al (2017). LncRNA UCA1 in anti-cancer drug re-sistance. Oncotarget, 8 (38): 64638- 64650.
25. Wang F, Li X, Xie X, et al (2008). UCA1, a non-protein-coding RNA up-regulated in bladder carcinoma and embryo, influenc-ing cell growth and promoting invasion. FEBS Lett, 582 (13): 1919-1927.
26. Xiao C, Wu CH, Hu HZ (2016). LncRNA UCA1 promotes epithelial-mesenchymal transition (EMT) of breast cancer cells via enhancing Wnt/beta-catenin signaling pathway. Eur Rev Med Pharmacol Sci, 20 (13): 2819-2824.
27. Li Y, Zeng Q, Qiu J, et al (2019). Long non-coding RNA UCA1 promotes breast cancer by upregulating PTP1B expression via inhibiting miR-206. Cancer Cell Int, 19:275.
28. Xin H, Liu N, Xu X, et al (2019). Knock-down of lncRNA‐UCA1 inhibits cell via-bility and migration of human glioma cells by miR‐193a‐mediated downregula-tion of CDK6. J Cell Biochem, 120 (9): 15157-15169.
29. Liu C, Jin J, Shi J, et al (2019). Long noncoding RNA UCA1 as a novel bi-omarker of lymph node metastasis and prognosis in human cancer: a meta-analysis. Biosci Rep, 39 (4): BSR20180995
30. Piri-Gharaghie T (2021). Polycystic ovary syndrome and genetic factors influencing its development: A review article. Pers M J, 6 (23): 25-29.
| Files | ||
| Issue | Vol 53 No 4 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v53i4.15571 | |
| Keywords | ||
| Long non-coding RNA Urothelial carcinoma associated 1 Cancer Knockdown miR-143 tumor-suppressor | ||
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How to Cite
1.
Montazeri-Najafabadi B, Doosti A, Kiani J. Long non-coding RNA UCA1 Knockdown Assisted by CRISPR/Cas9 in Female Cancer Cell Lines Increases Mir-143 Tumor-Suppressor. Iran J Public Health. 2024;53(4):934-946.
