AI Article Synopsis
- The study aimed to create a low expression cell line of DNMT1 in 16HBE cells to analyze its effects on the cell cycle and DNA methylation.
- The researchers employed Lenti-virus induced RNA interference to introduce various shRNA fragments and utilized flow cytometry and 5-mC immunofluorescence to assess the cell cycle and DNA methylation levels.
- Results showed a 44% decrease in DNMT1 protein levels without significant changes in the cell cycle or overall genomic DNA methylation in the modified cells compared to the control.
Article Abstract
Objective: To construct DNA methyltransferase 1 (DNMT1) low expression 16HBE cell line and observe the variation of cell cycle and global genomic DNA methylation.
Methods: The method of Lenti-virus induced RNA interference was applied to introduce four different shRNA fragment into 16HBE cells. Flow cytometry and 5-mC immunofluorescence methods were used to observe the cell cycle and global DNA methylation status of DNMT1 low expression 16HBE cells.
Results: The DNMT1 protein relative expression level of 16HBE-shDNMT1-4 cell line was down regulated about 44% (P < 0.05) compared with the control. No obvious differences of cell cycle and global genome DNA methylation status were observed between the 16HBE and 16HBE-shDNMT1.
Conclusion: The DNMT1 gene low expression cell is successfully constructed, and there are no obvious changes happened on the cell cycle and global genomic DNA methylation.
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