Epigenetic modulation of retinoic acid receptor beta2 by the histone deacetylase inhibitor MS-275 in human renal cell carcinoma.
OBJECTIVE
Histone deacetylase (HDAC) inhibitors have been shown to reverse epigenetic repression of certain genes, including retinoic acid receptor beta2 (RARbeta2). In this study, we examined whether RARbeta2 expression is repressed in human renal cell carcinoma (RCC) and whether the HDAC inhibitor MS-275 may revert its epigenetic repression.


METHODS
Six human tumor RCC cell lines were analyzed for RARbeta2 gene expression and for methylation and acetylation status at the promoter level. Modulation of RARbeta2 expression and correlation with antitumor activity by combination of MS-275 with 13-cis-retinoic acid (CRA) was assessed in a RARbeta2-negative RCC cell line.


RESULTS
RARbeta2 expression was either strongly present, weakly expressed, or absent in the RCC cell lines analyzed. Methylation-specific PCR indicated that the RARbeta2 promoter was partially methylated in three of the cell lines. CRA treatment did not inhibit clonogenic growth in the RARbeta2-negative cell line RCC1.18, whereas MS-275 induced a dose-dependent inhibitory effect. A greater inhibitory effect was observed with combination treatment (MS-275 + CRA). Treatment with MS-275 was associated with histone acetylation at the promoter level and synergistic gene reexpression of RARbeta2 in combination with CRA. RARbeta2 reexpression was associated with synergistic induction of the retinoid-responsive gene HOXA5. In vivo, single-agent CRA treatment showed no significant effect, whereas MS-275 and the combination induced a regression of RCC1.18 tumor xenografts. Discontinuation of treatment produced tumor recurrence in MS-275-treated mice, whereas animals treated with the combination remained tumor free.


CONCLUSIONS
The HDAC inhibitor MS-275 seems to revert retinoid resistance due to epigenetic silencing of RARbeta2 in a human RCC model and has greater antitumor activity in combination with CRA compared with single agents. Thus, the combination of HDAC inhibitors and retinoids may represent a novel therapeutic approach in patients with RCC.
Wang
Xiao-Fei
XF
Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, Maryland, USA.
Qian
David Z
DZ
Ren
Mingqiang
M
Kato
Yukihiko
Y
Wei
Yongfeng
Y
Zhang
Lu
L
Fansler
Zoya
Z
Clark
Doug
D
Nakanishi
Osamu
O
Pili
Roberto
R
D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D001549:Benzamides / Q000008:administration & dosage / Q000494:pharmacology*; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015474:Isotretinoin / Q000008:administration & dosage / Q000494:pharmacology; D007680:Kidney Neoplasms / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control; D008297:Male; D051379:Mice; D008819:Mice, Nude; D016513:Mice, SCID; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011725:Pyridines / Q000008:administration & dosage / Q000494:pharmacology*; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000235:genetics* / Q000378:metabolism; D000072482:Retinoic Acid Receptor alpha; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014212:Tretinoin / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays; D000097768:Retinoic Acid Receptor gamma
D001549:Benzamides; D056572:Histone Deacetylase Inhibitors; D018398:Homeodomain Proteins; D015474:Isotretinoin; D010750:Phosphoproteins; D011725:Pyridines; D012334:RNA, Neoplasm; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; D014212:Tretinoin; D000097768:Retinoic Acid Receptor gamma; C489213:HOXA5 protein, human; C000607071:RARA protein, human; C000607072:Rara protein, mouse; C082925:retinoic acid receptor beta; C118739:entinostat