17beta-hydroxysteroid dehydrogenase type 1 modulates breast cancer protein profile and impacts cell migration.
BACKGROUND
Human 17beta-hydroxysteroid dehydrogenase type 1 (17β-HSD1) is a steroid-converting enzyme that has long been known to play critical roles in estradiol synthesis and more recently in dihydrotestosterone (DHT) inactivation, showing a dual function that promotes breast cancer cell proliferation. Previously, we reported the first observation of the influence of the enzyme on endogenous estrogen-responsive gene expression. Here, we demonstrate the impact of 17β-HSD1 expression on the breast cancer cell proteome and investigate its role in cell migration.


METHODS
17β-HSD1 was stably transfected in MCF7 cells and the proteome of the generated cells overexpressing 17β-HSD1 (MCF7-17βHSD1 cells) was compared to that of the wild type MCF7 cells. Proteomics study was performed using two-dimensional gel electrophoresis followed by mass spectrometry analysis of differentially expressed protein spots. Reverse transcription quantitative real-time PCR (RT-qPCR) was used to investigate the transcription of individual gene. The effect of 17β-HSD1 on MCF7 cell migration was verified by a wound-healing assay.


RESULTS
Proteomic data demonstrate that the expression of more than 59 proteins is modulated following 17β-HSD1 overexpression. 17β-HSD1 regulates the expression of important genes and proteins that are relevant to cell growth control, such as BRCA2 and CDKN1A interacting protein (BCCIP) and proliferating cell nuclear antigen (PCNA) which are down- and upregulated in MCF7-17βHSD1 cells, respectively. RT-qPCR data reveal that 17β-HSD1 increases the mRNA levels of estrogen receptors (ER) alpha and beta by 171 and 120%, respectively, while decreasing that of the androgen receptor by 64%. Interestingly, 17β-HSD1 increases the mRNA transcript (by 3.6 times) and the protein expression of the metastasis suppressor gene nm23-H1 and the expression of the two enzymes are closely correlated. We have further shown that 17β-HSD1 expression is associated with an increase of MCF7 cell migration.


CONCLUSIONS
In addition to the regulation of important genes, we have demonstrated for the first time that 17β-HSD1 increases breast cancer cell migration, in spite of its positive regulation of the antimetastatic gene NM23. This is also correlated to its stimulation of breast cancer cell growth, further confirming its targeting in ER positive breast cancer. The novel findings in this study suggest several directions for future research on the contribution of 17β-HSD1 to breast cancer progression and related treatment.
Laboratory of Molecular Endocrinology and Oncology, Centre Hospitalier Universitaire de Québec Research Center (CHUQ - CHUL) and Department of Molecular Medicine, Laval University, 2705 boulevard Laurier, Québec G1V 4G2, Canada.
Juliette A
JA
Aka
Mouna
M
Zerradi
François
F
Houle
Jacques
J
Huot
Sheng-Xiang
SX
Lin
D024682:BRCA2 Protein / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D004960:Estradiol Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000235:genetics; D047629:Estrogen Receptor beta / Q000235:genetics; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D054778:NM23 Nucleoside Diphosphate Kinases / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D040081:Protein Array Analysis; D020543:Proteome / Q000032:analysis; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D011944:Receptors, Androgen / Q000235:genetics
C415366:BCCIP protein, human; D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D054778:NM23 Nucleoside Diphosphate Kinases; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D020543:Proteome; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011944:Receptors, Androgen; D004960:Estradiol Dehydrogenases; C513331:HSD17B1 protein, human; C517938:NME1 protein, human