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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 downregulates CCNDs.Notes: (A) mRNA levels of CCND1 and CCND2 were measured by qPCR in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (B) Western blot analysis of CCND1 and CCND2 protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots. Abbreviations: CCND1, cyclin D1; CCND2, cyclin D2; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
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f5-ott-10-1865: DNMT1 knockdown downregulates CCNDs.Notes: (A) mRNA levels of CCND1 and CCND2 were measured by qPCR in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (B) Western blot analysis of CCND1 and CCND2 protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots. Abbreviations: CCND1, cyclin D1; CCND2, cyclin D2; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Mentions: Furthermore, we found that mRNA and protein levels of CCND1 and CCND2 were significantly decreased in DNMT1 siRNA-transfected AN3CA cells compared to that in siRNA NC-transfected cells (Figure 5A and B). These data indicate that DNMT1 upregulates CCND1 and CCND2 to enhance the proliferation of EC cells.


DNMT1 regulates human endometrial carcinoma cell proliferation
DNMT1 knockdown downregulates CCNDs.Notes: (A) mRNA levels of CCND1 and CCND2 were measured by qPCR in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (B) Western blot analysis of CCND1 and CCND2 protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots. Abbreviations: CCND1, cyclin D1; CCND2, cyclin D2; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5384697&req=5

f5-ott-10-1865: DNMT1 knockdown downregulates CCNDs.Notes: (A) mRNA levels of CCND1 and CCND2 were measured by qPCR in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. (B) Western blot analysis of CCND1 and CCND2 protein levels in AN3CA cells transfected with DNMT1 siRNA or siRNA NC. GAPDH was the loading control. Shown are the representative blots. Abbreviations: CCND1, cyclin D1; CCND2, cyclin D2; PCR, polymerase chain reaction; qPCR, real-time quantitative PCR; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
Mentions: Furthermore, we found that mRNA and protein levels of CCND1 and CCND2 were significantly decreased in DNMT1 siRNA-transfected AN3CA cells compared to that in siRNA NC-transfected cells (Figure 5A and B). These data indicate that DNMT1 upregulates CCND1 and CCND2 to enhance the proliferation 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.