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SOX2OT knockdown derived changes in mitotic regulatory gene network of cancer cells
Marie Saghaeian Jazi1, Nader Mansour Samaei2, Seyed Javad Mowla3
11Metabolic Disorders Research Center, Golestan University of Medical Sciences, Gorgan, Iran.
Background:
SOX2 overlapping transcript (SOX2OT) is a long non-coding RNA, over-expressed in human tumor tissues and embryonic cells. Evidences support its function in the cell cycle; however there is no clear mechanism explaining its function in cell proliferation regulation. Here we investigated cancer cell response to SOX2OT knockdown by RNA sequencing.
Methods:
SOX2OT expression was inhibited by siRNA in two cancer cell lines (A549, U-87 MG), then the RNA of treated cells were used for the cDNA library synthesis and RNA sequencing. The differentially expressed genes were used for functional enrichment and the gene expression network was analyzed to find the most relevant biological process with SOX2OT function. Furthermore, the expression change of candidate genes was measured by qRT-PCR for more confirmation and the cell cycle was monitored by PI staining.
Results:
Our findings showed that SOX2OT knockdown affects the cellular gene expression generally with enriched cell proliferation and development biological process. Particularly, the cell cycle and mitotic regulatory genes expression including: CDK2, CDK2AP2, ACTR3, and chromosome structure associated genes like SMC4, INCENP and GNL3L are changed in treated cancer cells.
Conclusion:
Our results propound SOX2OT association with cell cycle and mitosis regulation in cancer cells.
Insights
SOX2 overlapping transcript (SOX2OT) knockdown impacts cancer cell gene expression, particularly affecting cell cycle and mitosis regulators. This research clarifies SOX2OT's role in cancer proliferation.
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Background:
- SOX2 overlapping transcript (SOX2OT) is a long non-coding RNA overexpressed in tumors.
- Its precise role in regulating cancer cell proliferation remains unclear.
- This study investigates SOX2OT's function in cancer cells.
Purpose of the Study:
- To elucidate the mechanism of SOX2OT in regulating cancer cell proliferation.
- To analyze gene expression changes following SOX2OT knockdown.
- To identify key biological processes influenced by SOX2OT.
Main Methods:
- SOX2OT expression was inhibited using siRNA in A549 and U-87 MG cancer cell lines.
- RNA sequencing was performed to analyze differential gene expression.
- Functional enrichment, gene network analysis, qRT-PCR, and cell cycle analysis (PI staining) were employed.
Main Results:
- SOX2OT knockdown broadly affected gene expression, enriching cell proliferation and development pathways.
- Expression of cell cycle and mitotic regulators (e.g., CDK2, SMC4) was altered.
- Chromosome structure-associated genes were also impacted.
Conclusions:
- SOX2OT is associated with the regulation of cell cycle and mitosis in cancer cells.
- These findings provide insights into SOX2OT's oncogenic functions.
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