Cholesterol Sulfonation Enzyme, SULT2B1b, Modulates AR and Cell Growth Properties in Prostate Cancer

Abstract

Cholesterol accumulates in prostate lesions and has been linked to prostate cancer (PCa) incidence and progression. However, how accumulated cholesterol contributes to PCa development and progression is not completely understood. Cholesterol sulfate (CS), the primary sulfonation product of cholesterol sulfotransferase (SULT2B1b), accumulates in human prostate adenocarcinoma and precancerous prostatic intraepithelial neoplasia (PIN) lesions compared to normal regions of the same tissue sample. Given the enhanced accumulation of CS in these lesions, it was hypothesized that SULT2B1b-mediated production of CS provides a growth advantage to these cells. To address this, PCa cells with RNAi-mediated knockdown (KD) of SULT2B1b were used to assess the impact on cell growth and survival. SULT2B1b is expressed and functional in a variety of prostate cells and the data demonstrate that SULT2B1b KD, in LNCaP and other androgen-responsive (VCaP and C4-2) cells, results in decreased cell growth/viability and induces cell death. SULT2B1b KD also decreases androgen receptor (AR) activity and expression at mRNA and protein levels. While AR overexpression has no impact on SULT2B1b KD-mediated cell death, addition of exogenous androgen is able to partially rescue the growth inhibition induced by SULT2B1b KD in LNCaP cells. These results suggest that SULT2B1b positively regulates the AR either through alterations in ligand availability or by interaction with critical co-regulators that influence AR activity. Implications These findings provide evidence that SULT2B1b is a novel regulator of AR activity and cell growth in prostate cancer and should be further investigated for therapeutic potential.

Targeting the Hsp90 C-terminal domain to induce allosteric inhibition and selective client downregulation

“…Cell lines were verified using cell line authentication testing from DDC Medical (Fairfield, OH) [50]. All experiments were completed within 20 passages of acquisition from ATCC.…”

Distinct expression patterns of SULT2B1b in human prostate epithelium

“…A stable cell line derived from LNCaP human prostate cancer cells that expresses doxycycline‐inducible SULT2B1b was used as a positive control for the antibody used for immunohistochemistry (IHC). As shown in Figure A, SULT2B1b staining intensity was markedly elevated in inducible cells treated with doxycycline, consistent with SULT2B1b induction previously shown by immunoblot . SULT2B1b IHC was then conducted on human prostate tissue samples.…”

“…Construction and characterization of LNCaP human prostate cancer cells with tetracycline‐inducible SULT2B1b have been previously described by our group . These cells were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium (HyClone, GE Healthcare, Uppsala, Sweden) supplemented with 10% fetal bovine serum (FBS; Corning, Corning, NY).…”

“…Total RNA was isolated using TRIzol Reagent (Thermo Fisher) according to the manufacturer's protocol. cDNA was synthesized with M‐MuLV Reverse Transcriptase (New England Biolabs, Ipswich, MA) as described previously . Quantitative real‐time polymerase chain reaction (qRT‐PCR) was conducted on LightCycler 96 (Roche, Basel, Switzerland) using PerfeCTa qPCR FastMix II (Quantabio, Beverly, MA) according to the manufacturer's protocol.…”

Review and perspectives on the applications of mass spectrometry imaging under ambient conditions

“…Furthermore, Banerjee et al investigated metabolites involved in the Krebs cycle in 64 tissue specimens, demonstrating the higher abundance of erythrose, glutamate, glucose and sorbitol, and lower abundance of succinate, salicylate, aconitate, and citrate in cancer tissue than in normal tissue . In another study conducted by Vickman et al on eight tissue samples, the spatial correlation between cholesterol sulfate and cholesterol sulfonation enzyme, SULT2B1b, was inspected within the prostate cancer tissue using DESI‐MS imaging . The colocalization of cholesterol sulfate and its associated enzyme in the tumour tissue corroborates a strong association of SULT2B1b activation with cell metabolism in prostate adenocarcinoma cells.…”