Regulation of Human Cytosolic Sulfotransferases 1C2 and 1C3 by Nuclear Signaling Pathways in LS180 Colorectal Adenocarcinoma Cells

Abstract

Cytosolic sulfotransferases (SULTs) catalyze the sulfate conjugation of a myriad of endogenous and xenobiotic substrates. Among the 13 human SULTs, little is known regarding regulation of the SULT1C subfamily. We evaluated the effects of a panel of transcription factor activators on levels of SULT1C mRNA (1C2 and 1C3) and protein (1C2) in LS180 colorectal adenocarcinoma cells. Treatment SULT1C2 protein content was strongly increased by 1,25(OH) 2 D 3 treatment and moderately increased by GW3965, GW4064, and rifampicin. To evaluate SULT1C2 transcriptional regulation, treatment effects were determined on reporter activity from transfected constructs containing ∼10 kb of the SULT1C2 gene. Treatment with GW3965, GW4064, or 1,25(OH) 2 D 3 increased reporter activity ∼2-, 5-, and 5.5-fold, respectively, from a construct containing mostly intron 1 of the SULT1C2 gene. Expression of AhR, LXRa, LXRb, PPARa, PPARg, PXR, and VDR was confirmed in LS180 cells using quantitative reverse-transcription polymerase chain reaction; however, FXR expression was negligible, suggesting that GW4064 increased SULT1C expression through an FXR-independent mechanism. Collectively, our findings are the first to characterize the regulation of human SULT1C2 and SULT1C3 expression by several transcription factor activators. Further, we determined that responsive regions for LXR and VDR are likely contained within intron 1 of the SULT1C2 gene.

Construction and analysis of three networks of genes and microRNAs in adenocarcinoma

“…These associated genes were a component of the associated gene network in adenocarcinoma. Finally, the last component of the associated gene network in adenocarcinoma was identified using the relevant literature (37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47). The four aforementioned components were then summarized as the associated gene network in adenocarcinoma.…”

Transcriptional Regulation of Cytosolic Sulfotransferase 1C2 by Vitamin D Receptor in LS180 Human Colorectal Adenocarcinoma Cells

“…LS180 colorectal adenocarcinoma cells were obtained from the American Type Culture Collection (Manassas, VA) and were cultured as previously described elsewhere (Rondini et al, 2014 s This article has supplemental material available at dmd.aspetjournals.org.…”

“…LS180 colorectal adenocarcinoma cells were obtained from the American Type Culture Collection (Manassas, VA) and were cultured as previously described elsewhere (Rondini et al, 2014). Human embryonic kidney 293 (HEK293) cells used for protein expression were obtained from Dr. Ye-Shih Ho (Wayne State University, Detroit, MI) and were cultured in Dulbecco's modified Eagle medium supplemented with 100 U/ml penicillin, 100 mg/ml streptomycin, and 10% fetal bovine serum (all purchased from Life Technologies, Grand Island, NY).…”

“…We recently reported that treatment of LS180 human colorectal adenocarcinoma cells with several nuclear receptor activators, including 3-[3-[N-(2-chloro-3-trifluoromethylbenzyl)-(2,2-diphenylethyl)amino] propyloxy]phenylacetic acid (GW3965, liver X receptor agonist), 3-(2,6-dichlorophenyl)-4-(39-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole (GW4064, farnesoid X receptor agonist), rifampicin (pregnane X receptor [PXR] agonist), or 1a,25-dihydroxyvitamin D 3 (VitD 3 , vitamin D receptor agonist), increased SULT1C2 expression (Rondini et al, 2014). The VitD 3 -mediated induction was especially pronounced and occurred at the mRNA, protein, and promoter activation levels.…”

“…Human SULT1C2 is expressed in multiple tissues, including fetal liver, stomach, and the vitamin D target tissues thyroid, intestine, and kidney (Runge-Morris and Kocarek, 2013). SULT1C2 does not metabolize prototypical SULT substrates but has been shown to sulfonate thyroid hormones and some phenols and to bioactivate the procarcinogen N-hydroxy-2-acetylaminofluorene (Sakakibara et al, 1998;Li et al, 2000;Allali-Hassani et al, 2007).We recently reported that treatment of LS180 human colorectal adenocarcinoma cells with several nuclear receptor activators, including 3-[3-[N-(2-chloro-3-trifluoromethylbenzyl)-(2,2-diphenylethyl)amino] propyloxy]phenylacetic acid (GW3965, liver X receptor agonist), 3-(2,6-dichlorophenyl)-4-(39-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole (GW4064, farnesoid X receptor agonist), rifampicin (pregnane X receptor [PXR] agonist), or 1a,25-dihydroxyvitamin D 3 (VitD 3 , vitamin D receptor agonist), increased SULT1C2 expression (Rondini et al, 2014). The VitD 3 -mediated induction was especially pronounced and occurred at the mRNA, protein, and promoter activation levels.…”