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The Impact of Si-SETDB1 on Migration and Invasion of Lung Adenocarcinoma Cells through SPG20 Methylation |
ZHAO Baoshan, et al |
The Affiliated Hospital of Chengde Medical University, Hebei Chengde 067000, China |
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Abstract Objective: To investigate the methylation levels of SETDB1 and SPG20 in lung adenocarcinoma and explore the impact of downregulating SETDB1 on SPG20 methylation, A549 cell migration, and invasion. Methods: Sixty cases of lung adenocarcinoma tissues and normal lung tissues were selected. qRT-PCR was used to detect mRNA relative expression levels, immunohistochemistry and western blot were used to detect protein expression levels, and gene methylation levels were assessed by bisulfite sequencing. Small interfering RNA (siRNA) targeting SETDB1 was constructed and transfected into cells using liposomes. qRT-PCR, Western blot, bisulfite sequencing, MTT assay, scratch healing assay, and Transwell chamber assay were performed to evaluate mRNA and protein expression, gene methylation, cell proliferation, migration, and invasion, respectively. Results: qRT-PCR results showed that SPG20 expression was downregulated (P < 0.05), while SETDB1 expression was upregulated (P < 0.05) in lung adenocarcinoma tissues. Western blot results indicated that SETDB1 protein expression in cancer tissues was significantly higher than in normal lung tissues, whereas SPG20 protein exhibited lower expression in cancer tissues, lower than in normal lung tissues. In cancer tissues, the SPG20 gene methylation rate was significantly higher than in normal lung tissues, with a statistically significant difference. SPG20 methylation level was positively correlated with SETDB1 expression. SETDB1 was highly expressed, while SPG20 was lowly expressed in lung adenocarcinoma cells. Conversely, in normal bronchial epithelial cells, SETDB1 was lowly expressed, and SPG20 was highly expressed. RNA interference with SETDB1 significantly reduced SETDB1 mRNA and protein expression in A549 cells, decreased SPG20 gene methylation, and inhibited cell proliferation, migration, and invasion. Conclusion: There is a correlation between methyltransferase SETDB1 and SPG20 methylation in lung adenocarcinoma. SETDB1 may serve as a catalytic enzyme for SPG20 methylation.
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