Efflux excretion of bisdemethoxycurcumin‐O‐glucuronide in UGT1A1‐overexpressing HeLa cells: Identification of breast cancer resistance protein (BCRP) and multidrug resistance‐associated proteins 1 (MRP1) as the glucuronide transporters

Abstract

Bisdemethoxycurcumin (BDMC) was a natural curcuminoid with many bioactivities present in turmeric (Curcuma longa L.). However, the disposition mechanisms of BDMC via uridine 5 0 -diphospho-glucuronosyltransferase (UGT) metabolism still remain unclear. Therefore, we aimed to determine the potential efflux transporters for the excretion of BDMC-O-glucuronide. Herein, chemical inhibition assays (Ko143, MK571, dipyridamole, and leukotriene C4) and biological inhibition experiments including stable knocked-down of breast cancer resistance protein (BCRP), multidrug resistance-associated proteins (MRPs) transporters were both performed in a HeLa cell line stably overexpressing UGT1A1 established previously. The results indicated that Ko143 (5 and 20 μM) caused a marked reduction in excretion rate (18.4-55.6%) and elevation of intracellular BDMC-O-glucuronide (28.8-48.1%), whereas MK-571 (5 and 20 μM) resulted in a significant decrease in excretion rate (6.2-61.6%) and increase of intracellular BDMC-O-glucuronide (maximal 27.1-32.6%). Furthermore, shRNAmediated silencing of BCRP transporter led to a marked reduction in the excretion rate (21.1-36.9%) and an obvious elevation of intracellular glucuronide (24.9%). Similar results were observed when MRP1 was partially silenced. In addition, MRP3 and MRP4 silencing both displayed no obvious changes on the excretion rate and intracellular levels of glucuronide. In conclusion, chemical inhibition and gene silencing results both indicated that generated BDMC-O-glucoside were excreted primarily by the BCRP and MRP1 transporters.

Metabolism and disposition of corylifol A from Psoralea corylifolia: metabolite mapping, isozyme contribution, species differences and identification of efflux transporters for corylifol A-O-glucuronide in HeLa1A1 cells

“…HeLa1A1 cells were established and validated as described previously Yang et al, 2018a;Zhang et al, 2019). Before excretion experiments, HeLa1A1 cells were cultured in DMEM containing 10% FBS, penicillin (100 U/mL) and streptomycin (100 μg/mL), maintaining at 37 ℃ in a humidified incubator with 5% CO 2 .…”

“…This excretion process needs to rely on the roles of efflux transporters, such as multidrug A c c e p t e d M a n u s c r i p t resistance-associated proteins (MRPs) and breast cancer resistance protein (BCRP) Qin et al, 2019;Zhang et al, 2015). Previous studies have indicated that many compounds conjugated with glucuronide were the substrates of MRPs and BCRP transporters (Yang et al, 2018a;Zhang et al, 2019). MRPs and BCRP also play an important role in determining the oral bioavailability and pharmacokinetics of the aglycones undergoing glucuronidation.…”

Study on mechanism of elemene reversing tumor multidrug resistance based on luminescence pharmacokinetics in tumor cells in vitro and in vivo

“…13 Comparatively, some traditional Chinese medicines (TCMs) have shown a signicant MDR reversal effect with fewer side effects. [14][15][16] In our former studies, we found that several TCMs-such as elemene (ELE), a noncytotoxic broad-spectrum anti-tumor drug extracted from Curcuma zedoaria Roscoe have great reversal effect in MDR cells. However, these TCMs do not always reduce the expression of MDR-related ABC transporters.…”

Potential metabolism determinants and drug–drug interactions of a natural flavanone bavachinin

“…43 A remarkable limitation is the unexplained role of MRP2 in excretion of G1 due to the absence of MRP2 transporters in HeLa1A1 cells. [21][22][23]25,41,43 Furthermore, MDCKII-MRP2-UGT1A1 cells could well characterize the glucuronidation by UGT1A1 and glucuronides excretion by MRP2 transporters. 45 In addition, Mrp2 (Abcc2) knock-out mice were usually used to evaluate the roles of Mrp2 according to the pharmacokinetic behaviors.…”

“…21 In our previous works, many glucuronides of natural compounds were the substrates of BCRP or MRPs transporters. [21][22][23][24][25] In addition, the glucuronidation activity would be altered when the function of BCRP or MRPs transporters were inhibited due to the presence of "glucuronidationtransport interplay". 21 However, the effects of BCRP and MRPs transporters on the metabolism and disposition of bavachinin still remains unknown.…”