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DNMT1 regulates human endometrial carcinoma cell proliferation

View Article: PubMed Central - PubMed

ABSTRACT

Endometrial carcinoma (EC) is the most common gynecologic malignancy, but the molecular events involved in the development and progression of EC remain unclear. This study aimed to investigate the role of DNA methyltransferase 1 (DNMT1), a member of DNA methyltransferases, in EC. AN3CA cells were transfected with DNMT1 siRNA. The proliferation, cell cycle, and apoptosis of AN3CA cells were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry. The expression of related genes was detected by polymerase chain reaction and Western blot analysis. Knockdown of DNMT1 inhibited the proliferation, induced apoptosis, and G0/G1 phase arrest of AN3CA cells. Furthermore, knockdown of DNMT1 upregulated the expression of nuclear factor kappa-B-inhibitor alpha (NF-κBIA) and Bax and downregulated the expression of Bcl-2 and CCND1/2 in AN3CA cells. In conclusion, this study provides the first evidence that knockdown of DNMT1 affects the expression of cell cycle- and apoptosis-associated proteins in EC cells, suggesting the potential of DNMT1 in EC therapy.

No MeSH data available.


DNMT1 knockdown alters the expression of apoptosis-related proteins.Notes: qPCR (A) and Western blot (B) analyses of NF-κBIA and p65 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (C) Western blot analysis of Caspase-2 protein level in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. qPCR (D) and Western blot (E) analyses of Bax and Bcl-2 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots.Abbreviations: NF-κBIA, nuclear factor kappa-B-inhibitor alpha; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR.
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f4-ott-10-1865: DNMT1 knockdown alters the expression of apoptosis-related proteins.Notes: qPCR (A) and Western blot (B) analyses of NF-κBIA and p65 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (C) Western blot analysis of Caspase-2 protein level in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. qPCR (D) and Western blot (E) analyses of Bax and Bcl-2 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots.Abbreviations: NF-κBIA, nuclear factor kappa-B-inhibitor alpha; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR.

Mentions: To understand the molecular mechanism by which DNMT1 modulates EC cell cycle and apoptosis, we detected the expression of a variety of proteins involved in cell cycle and apoptosis. qPCR and Western blot analyses showed that NF-κBIA and Caspase-2 were upregulated and p65 was downregulated in DNMT1 siRNA-transfected AN3CA cells (Figure 4A–C). Moreover, Bax was upregulated and Bcl-2 was downregulated in AN3CA cells after the knockdown of DNMT1 (Figure 4D and E). These results reveal that DNMT1 regulates these apoptosis-related genes to promote the apoptosis of EC cells.


DNMT1 regulates human endometrial carcinoma cell proliferation
DNMT1 knockdown alters the expression of apoptosis-related proteins.Notes: qPCR (A) and Western blot (B) analyses of NF-κBIA and p65 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (C) Western blot analysis of Caspase-2 protein level in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. qPCR (D) and Western blot (E) analyses of Bax and Bcl-2 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots.Abbreviations: NF-κBIA, nuclear factor kappa-B-inhibitor alpha; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5384697&req=5

f4-ott-10-1865: DNMT1 knockdown alters the expression of apoptosis-related proteins.Notes: qPCR (A) and Western blot (B) analyses of NF-κBIA and p65 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (C) Western blot analysis of Caspase-2 protein level in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. qPCR (D) and Western blot (E) analyses of Bax and Bcl-2 mRNA and protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots.Abbreviations: NF-κBIA, nuclear factor kappa-B-inhibitor alpha; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR.
Mentions: To understand the molecular mechanism by which DNMT1 modulates EC cell cycle and apoptosis, we detected the expression of a variety of proteins involved in cell cycle and apoptosis. qPCR and Western blot analyses showed that NF-κBIA and Caspase-2 were upregulated and p65 was downregulated in DNMT1 siRNA-transfected AN3CA cells (Figure 4A–C). Moreover, Bax was upregulated and Bcl-2 was downregulated in AN3CA cells after the knockdown of DNMT1 (Figure 4D and E). These results reveal that DNMT1 regulates these apoptosis-related genes to promote the apoptosis of EC cells.

View Article: PubMed Central - PubMed

ABSTRACT

Endometrial carcinoma (EC) is the most common gynecologic malignancy, but the molecular events involved in the development and progression of EC remain unclear. This study aimed to investigate the role of DNA methyltransferase 1 (DNMT1), a member of DNA methyltransferases, in EC. AN3CA cells were transfected with DNMT1 siRNA. The proliferation, cell cycle, and apoptosis of AN3CA cells were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry. The expression of related genes was detected by polymerase chain reaction and Western blot analysis. Knockdown of DNMT1 inhibited the proliferation, induced apoptosis, and G0/G1 phase arrest of AN3CA cells. Furthermore, knockdown of DNMT1 upregulated the expression of nuclear factor kappa-B-inhibitor alpha (NF-κBIA) and Bax and downregulated the expression of Bcl-2 and CCND1/2 in AN3CA cells. In conclusion, this study provides the first evidence that knockdown of DNMT1 affects the expression of cell cycle- and apoptosis-associated proteins in EC cells, suggesting the potential of DNMT1 in EC therapy.

No MeSH data available.