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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.


Related in: MedlinePlus

DNMT1 knockdown induces cell cycle arrest and apoptosis of AN3CA cells.Notes: (A) Knockdown of DNMT1 induced cell cycle arrest in G0/G1 phase at 5 days after the transfection of DNMT1 siRNA. (B) Percentage of cells at G0 + G1, and S phases. The data presented have been background subtracted using data from unstained cells. (C) Flow cytometry analysis of AN3CA cells transfected with DNMT1 siRNA or siRNA NC. The lower right quadrants of the histograms indicate the percentage of early apoptotic cells. (D) Percentage of apoptotic cells. The data presented have been background subtracted using data from unstained cells.Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate; NC, negative control; Ph, phase.
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f3-ott-10-1865: DNMT1 knockdown induces cell cycle arrest and apoptosis of AN3CA cells.Notes: (A) Knockdown of DNMT1 induced cell cycle arrest in G0/G1 phase at 5 days after the transfection of DNMT1 siRNA. (B) Percentage of cells at G0 + G1, and S phases. The data presented have been background subtracted using data from unstained cells. (C) Flow cytometry analysis of AN3CA cells transfected with DNMT1 siRNA or siRNA NC. The lower right quadrants of the histograms indicate the percentage of early apoptotic cells. (D) Percentage of apoptotic cells. The data presented have been background subtracted using data from unstained cells.Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate; NC, negative control; Ph, phase.

Mentions: To explore the possible mechanisms by which DNMT1 promotes EC cell proliferation, the effect of DNMT1 silencing on cell cycle progression of AN3CA cells was analyzed by flow cytometry. The results showed that knockdown of DNMT1 in AN3CA cells induced an increase in the percentage of cells in G0/G1 phase (from 82.07%±0.80% in the siRNA NC group to 85.77%±0.29% in the DNMT1 siRNA group, P<0.05), parallel with a decrease in the percentage of cells in S phase (from 17.17%±0.59% in siRNA NC group to 13.87%±0.40% in DNMT1 siRNA group, P<0.05) (Figure 3A and B). These results suggest that DNMT1 knockdown inhibits EC cell proliferation through inducing G0/G1 cell cycle arrest.


DNMT1 regulates human endometrial carcinoma cell proliferation
DNMT1 knockdown induces cell cycle arrest and apoptosis of AN3CA cells.Notes: (A) Knockdown of DNMT1 induced cell cycle arrest in G0/G1 phase at 5 days after the transfection of DNMT1 siRNA. (B) Percentage of cells at G0 + G1, and S phases. The data presented have been background subtracted using data from unstained cells. (C) Flow cytometry analysis of AN3CA cells transfected with DNMT1 siRNA or siRNA NC. The lower right quadrants of the histograms indicate the percentage of early apoptotic cells. (D) Percentage of apoptotic cells. The data presented have been background subtracted using data from unstained cells.Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate; NC, negative control; Ph, phase.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5384697&req=5

f3-ott-10-1865: DNMT1 knockdown induces cell cycle arrest and apoptosis of AN3CA cells.Notes: (A) Knockdown of DNMT1 induced cell cycle arrest in G0/G1 phase at 5 days after the transfection of DNMT1 siRNA. (B) Percentage of cells at G0 + G1, and S phases. The data presented have been background subtracted using data from unstained cells. (C) Flow cytometry analysis of AN3CA cells transfected with DNMT1 siRNA or siRNA NC. The lower right quadrants of the histograms indicate the percentage of early apoptotic cells. (D) Percentage of apoptotic cells. The data presented have been background subtracted using data from unstained cells.Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate; NC, negative control; Ph, phase.
Mentions: To explore the possible mechanisms by which DNMT1 promotes EC cell proliferation, the effect of DNMT1 silencing on cell cycle progression of AN3CA cells was analyzed by flow cytometry. The results showed that knockdown of DNMT1 in AN3CA cells induced an increase in the percentage of cells in G0/G1 phase (from 82.07%±0.80% in the siRNA NC group to 85.77%±0.29% in the DNMT1 siRNA group, P<0.05), parallel with a decrease in the percentage of cells in S phase (from 17.17%±0.59% in siRNA NC group to 13.87%±0.40% in DNMT1 siRNA group, P<0.05) (Figure 3A and B). These results suggest that DNMT1 knockdown inhibits EC cell proliferation through inducing G0/G1 cell cycle arrest.

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-&kappa;BIA) 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.


Related in: MedlinePlus