scholarly journals CRISPR/Cas9-Mediated Gene Knockout of ARID1A Promotes Primary Progesterone Resistance by Downregulating Progesterone Receptor B in Endometrial Cancer Cells

2019 ◽  
Vol 27 (9) ◽  
pp. 1051-1060 ◽  
Author(s):  
Haizhen Wang ◽  
Zhenghua Tang ◽  
Ting Li ◽  
Menglu Liu ◽  
Yong Li ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Transcriptional Activity ◽  
Gene Knockout ◽  
Cell Counting ◽  
Akt Inhibitor ◽  
Akt Activation ◽  
Progesterone Resistance ◽  
Ishikawa Cells ◽  
Progesterone Receptor B

Medroxyprogesterone (MPA) is used for the conservative treatment of endometrial cancer. Unfortunately, progesterone resistance seriously affects its therapeutic effect. The purpose of the current study was to investigate the influence of deletion of AT-rich interactive domain 1A (ARID1A) in progesterone resistance in Ishikawa cells. Ablation of ARID1A was conducted through the CRISPR/Cas9 technology. Acquired progesterone-resistant Ishikawa (Ishikawa-PR) cells were generated by chronic exposure of Ishikawa cells to MPA. The sensitivity of the parental Ishikawa, Ishikawa-PR, and ARID1A-deficient cells to MPA and/or LY294002 was determined using the Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis. In addition, Western blot analysis and reverse transcription-polymerase chain reaction was performed to evaluate the mRNA and protein expression levels of ARID1A, progesterone receptor B (PRB), and P-AKT. Both Ishikawa-PR and ARID1A knockout cells showed insensitivity to MPA, downregulation of PRB, and hyperphosphorylation of AKT compared to the parental Ishikawa cells. Pretreatment with LY294002 significantly enhanced the ability of MPA to suppress proliferation and to induce apoptosis in the parental and Ishikawa-PR cells via the inhibition of AKT activation and upregulation of PRB transcriptional activity. However, the PRB transcriptional activity and insensitivity to MPA were irreversible by LY294002 in ARID1A-deficient cells. Ablation of ARID1A is associated with low PRB expression, which serves an important role in primary progesterone resistance. Akt inhibition cannot rescue PRB or sensitize to MPA in ARID1A knockout cells. These findings suggest that ARID1A may act as a reliable biomarker to predict the response for the combination of AKT inhibitor and MPA treatment.

scholarly journals Akt regulates progesterone receptor B-dependent transcription and angiogenesis in endometrial cancer cells

Oncogene ◽  
2016 ◽  
Vol 35 (39) ◽  
pp. 5191-5201 ◽  
Author(s):  
I I Lee ◽  
K Maniar ◽  
J P Lydon ◽  
J J Kim
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Cancer Cells ◽  
Progesterone Receptor B

Megestrol acetate drives endometrial carcinoma cell senescence via interacting with progesterone receptor B/FOXO1 axis

2021 ◽  
pp. 153537022110265
Author(s):  
Hong Wang ◽  
Huirong Shi
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Cancer Cell ◽  
Cell Senescence ◽  
Megestrol Acetate ◽  
G1 Arrest ◽  
Cancer Cell Growth ◽  
Endometrial Cancer Cell ◽  
Cycle Arrest ◽  
Progesterone Receptor B

Megestrol acetate is a common and efficient anticancer progesterone. To explore the activity and the therapeutic mechanisms of megestrol acetate in endometrial cancer, human endometrial cancer cell lines Ishikawa and HHUA overexpressing progesterone receptor A (PR-A) and progesterone receptor B (PR-B) were treated with megestrol acetate. Cell viability, apoptosis, cycle arrest, and senescence, as well as the expressions of p21 and p16, two hallmarks of cellular senescence, were evaluated. Compared with the control, >10 nmol/L megestrol acetate treatment could significantly reduce endometrial cancer cell growth, and induce the irreversible G1 arrest and cell senescence. The expression of cyclin D1 in megestrol acetate treated cells was downregulated, while the expressions of p21 and p16 were upregulated via PR-B isoform. FOXO1 inhibitor AS1842856 could significantly abrogate megestrol acetate-induced cell senescence, suggesting that FOXO1 was involved in megestrol acetate/PR-B axis. These findings may provide a new understanding for the treatment of human endometrial cancer.

scholarly journals The Regulation and Function of the Forkhead Transcription Factor, Forkhead Box O1, Is Dependent on the Progesterone Receptor in Endometrial Carcinoma

Endocrinology ◽  
2007 ◽  
Vol 149 (4) ◽  
pp. 1942-1950 ◽  
Author(s):  
Erin C. Ward ◽  
Anna V. Hoekstra ◽  
Leen J. Blok ◽  
P. Hanifi-Moghaddam ◽  
John R. Lurain ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Cell Lines ◽  
Type I ◽  
Ishikawa Cell ◽  
Protein Levels ◽  
Forkhead Box ◽  
Ishikawa Cells ◽  
Presence And Absence ◽  
And Function

In many type I endometrial cancers, the PTEN gene is inactivated, which ultimately leads to constitutively active Akt and the inhibition of Forkhead box O1 (FOXO1), a member of the FOXO subfamily of Forkhead/winged helix family of transcription factors. The expression, regulation, and function of FOXO1 in endometrial cancer were investigated in this study. Immunohistochemical analysis of 49 endometrial tumor tissues revealed a decrease of FOXO1 expression in 95.9% of the cases compared with the expression in normal endometrium. In four different endometrial cancer cell lines (ECC1, Hec1B, Ishikawa, and RL95), FOXO1 mRNA was expressed at similar levels; however, protein levels were low or undetectable in Ecc1, Ishikawa, and RL95 cells. Using small interfering RNA technology, we demonstrated that the low levels of FOXO1 protein were due to the involvement of Skp2, an oncogenic subunit of the Skp1/Cul1/F-box protein ubiquitin complex, given that silencing Skp2 increased FOXO1 protein expression in Ishikawa cells. Inhibition of Akt in Ishikawa cells also increased nuclear FOXO1 protein levels. Additionally, progestins increased FOXO1 protein levels, specifically through progesterone receptor B (PRB) as determined by using stably transfected PRA-specific and PRB-specific Ishikawa cell lines. Finally, overexpression of triple mutant (Tm) FOXO1 in the PR-specific Ishikawa cell lines caused cell cycle arrest and significantly decreased proliferation in the presence and absence of the progestin, R5020. Furthermore, TmFOXO1 overexpression induced apoptosis in PRB-specific cells in the presence and absence of ligand. Taken together, these data provide insight into the phosphoinositide-3-kinase/Akt/FOXO pathway for the determination of progestin responsiveness and the development of alternate therapies for endometrial cancer.

scholarly journals Progesterone Receptor-B Induction of BIRC3 Protects Endometrial Cancer Cells from AP1-59-Mediated Apoptosis

2011 ◽  
Vol 2 (3) ◽  
pp. 170-181 ◽  
Author(s):  
Nikki L. Neubauer ◽  
Erin C. Ward ◽  
Parin Patel ◽  
Zhenxiao Lu ◽  
Irene Lee ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Cancer Cells ◽  
Progesterone Receptor B

Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines

2005 ◽  
Vol 99 (1) ◽  
pp. 135-141 ◽  
Author(s):  
Yuning Xiong ◽  
Sean C. Dowdy ◽  
Jesus Gonzalez Bosquet ◽  
Ying Zhao ◽  
Norman L. Eberhardt ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Endometrial Cancer Cell ◽  
Progesterone Receptor B

scholarly journals PFKFB4 promotes lung adenocarcinoma progression via phosphorylating and activating transcriptional coactivator SRC-2

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jiguang Meng ◽  
Xuxin Chen ◽  
Zhihai Han
Keyword(s):  
Lung Adenocarcinoma ◽  
Transcriptional Activity ◽  
Cell Counting ◽  
Migration And Invasion ◽  
Transcriptional Coactivator ◽  
Downstream Target ◽  
Steroid Receptor Coactivator ◽  
Adenocarcinoma Cells ◽  
Family Protein ◽  
Lung Adenocarcinoma Cells

Abstract Background To investigate the role and its potential mechanism of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4) in lung adenocarcinoma. Methods Co-immunoprecipitation was performed to analyze the interaction between PFKFB4 and SRC-2. Western blot was used to investigate the phosphorylation of steroid receptor coactivator-2 (SRC-2) on the condition that PFKFB4 was knockdown. Transcriptome sequencing was performed to find the downstream target of SRC-2. Cell Counting Kit-8 (CCK-8) assay, transwell assay and transwell-matrigel assay were used to examine the proliferation, migration and invasion abilities in A549 and NCI-H1975 cells with different treatment. Results In our study we found that PFKFB4 was overexpressed in lung adenocarcinoma associated with SRC family protein and had an interaction with SRC-2. PFKFB4 could phosphorylate SRC-2 at Ser487, which altered SRC-2 transcriptional activity. Functionally, PFKFB4 promoted lung adenocarcinoma cells proliferation, migration and invasion by phosphorylating SRC-2. Furthermore, we identified that CARM1 was transcriptionally regulated by SRC-2 and involved in PFKFB4-SRC-2 axis on lung adenocarcinoma progression. Conclusions Our research reveal that PFKFB4 promotes lung adenocarcinoma cells proliferation, migration and invasion via enhancing phosphorylated SRC-2-mediated CARM1 expression.

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