Volume 8, Issue 2 (7-2021)                   J Jiroft Univ Med Sci 2021, 8(2): 652-663 | Back to browse issues page

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Azadbakht N, Doosti A, Jami M. Editing of LINC00511 gene with a new CRISPR/Cas9 technique and evaluation of its effects on lung cancer cell line. J Jiroft Univ Med Sci 2021; 8 (2) :652-663
URL: http://journal.jmu.ac.ir/article-1-501-en.html
1- PhD Student at Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
2- Associate Professor at Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran , abbasdoosti@yahoo.com
3- Assistant Professor at Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran , Assistant Professor at Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
Abstract:   (1595 Views)
Introduction:  Lung cancer is the second most common cancer in the world. The COR-L105 cell line is used as one of the most popular lung cancer cell lines in various studies. Long non-coding RNAs (lncRNAs) are a group of important cellular factors that play a key role in many processes such as growth, differentiation, replication, transcription, translation. LncRNA LINC00511 is a transcriptional regulator and has been reported to increase its expression in various cancers. The aim of this study was to knockout the LINC00511 gene using a CRISPR/Cas9 in COR-L105 cells and to investigate its effects on cancer progression and apoptosis.
Methods:  In this experimental study, two sgRNAs were designed for LINC00511 gene and cloned into two CRISPR vectors, separately. Two sgRNAs vectors were transferred to COR-L105 cells using lipofectamine. After confirmation of LINC00511 gene knockout from target cells, changes in cell proliferation and apoptosis were assayed by MTT and flow cytometry. Furthermore, the expression of genes associated with apoptosis and tumorigenesis was examined by real-time PCR.
Results: LINC00511 gene knockout from COR-L105 cell line was performed. The expression of BCL2, survivin, EZH2, c-MYC, and MAX genes showed a meaningful decrease related to the expression in treated cells (edited) compared to the control cells (p˂0.05). Increased expression of p21 and p57 genes was also seen in the edited cells (p˂0.05). Decreased cell proliferation and increased apoptosis were observed in manipulated cells.
Conclusion: Knocking of LINC00511 gene in lung cancer cell line caused apoptosis and decreased cell proliferation. Therefore, inhibiting the expression of the LINC00511 gene in cancer cells leads to the control their proliferation.
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Type of Study: Research | Subject: Medical Sciences / Medical Genetics
Received: 2021/06/6 | Accepted: 2021/09/6 | Published: 2021/09/20

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