Nuclear versus cytoplasmic localization of filamin A in prostate cancer: immunohistochemical correlation with metastases.
OBJECTIVE
We previously showed that nuclear localization of the actin-binding protein, filamin A (FlnA), corresponded to hormone-dependence in prostate cancer. Intact FlnA (280 kDa, cytoplasmic) cleaved to a 90 kDa fragment which translocated to the nucleus in hormone-naïve cells, whereas in hormone-refractory cells, FlnA was phosphorylated, preventing its cleavage and nuclear translocation. We have examined whether FlnA localization determines a propensity to metastasis in advanced androgen-independent prostate cancer.


METHODS
We examined, by immunohistochemistry, FlnA localization in paraffin-embedded human prostate tissue representing different stages of progression. Results were correlated with in vitro studies in a cell model of prostate cancer.


RESULTS
Nuclear FlnA was significantly higher in benign prostate (0.6612 +/- 0.5888), prostatic intraepithelial neoplasia (PIN; 0.6024 +/- 0.4620), and clinically localized cancers (0.69134 +/- 0.5686) compared with metastatic prostate cancers (0.3719 +/- 0.4992, P = 0.0007). Cytoplasmic FlnA increased from benign prostate (0.0833 +/- 0.2677), PIN (0.1409 +/- 0.2293), localized cancers (0.3008 +/- 0.3762, P = 0.0150), to metastases (0.7632 +/- 0.4414, P < 0.00001). Logistic regression of metastatic versus nonmetastatic tissue yielded the area under the receiver operating curve as 0.67 for nuclear-FlnA, 0.79 for cytoplasmic-FlnA, and 0.82 for both, indicating that metastasis correlates with cytoplasmic to nuclear translocation. In vitro studies showed that cytoplasmic localization of FlnA induced cell invasion whereas nuclear translocation of the protein inhibited it. FlnA dephosphorylation with the protein kinase A inhibitor H-89 facilitated FlnA nuclear translocation, resulting in decreased invasiveness and AR transcriptional activity, and induced sensitivity to androgen withdrawal in hormone-refractory cells.


CONCLUSIONS
The data presented in this study indicate that in prostate cancer, metastasis correlates with cytoplasmic localization of FlnA and may be prevented by cleavage and subsequent nuclear translocation of this protein.
Bedolla
Roble G
RG
University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA.
Wang
Yu
Y
Asuncion
Alfredo
A
Chamie
Karim
K
Siddiqui
Salma
S
Mudryj
Maria M
MM
Prihoda
Thomas J
TJ
Siddiqui
Javed
J
Chinnaiyan
Arul M
AM
Mehra
Rohit
R
de Vere White
Ralph W
RW
Ghosh
Paramita M
PM
D000726:Androgen Antagonists / Q000627:therapeutic use; D045744:Cell Line, Tumor; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism*; D003285:Contractile Proteins / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm; D064448:Filamins; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D046888:Tissue Array Analysis
D000726:Androgen Antagonists; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins