Molecular expression and functional activity of sodium dependent multivitamin transporter in human prostate cancer cells.
Nutrient transporters expressed on cell membrane have been targeted for enhancing bioavailability of poorly permeable drugs. Sodium dependent multivitamin transporter (SMVT) is once such carrier system, utilized for improving drug targeting to specific tissues. Therefore, the main objective of this study is to characterize SMVT in human derived prostate cancer cells (PC-3). Reverse transcription polymerase chain reaction (RT-PCR) analysis has provided product band at 774 bp, specific to SMVT. The mechanism and intracellular regulation of [3H]-biotin is also studied. [3H]-biotin uptake is found to be time and concentration dependent with K(m) and V(max) values of 19±2 μM and 23±1 pmol/min/mg protein, respectively. The uptake process is saturable in micromolar concentration range but linear in nanomolar concentration range. [3H]-biotin uptake shows significant sodium, temperature, pH and energy dependency. The process is strongly inhibited by unlabeled biotin and structural analogs such as desthiobiotin, pantothenate, lipoate and valeric acid. Intracellular regulatory pathways such as Ca(2+)/calmodulin and PKC pathway but not PTK pathway appears to play an important role in modulating [3H]-biotin uptake. This study for the first time confirms the molecular expression of SMVT and demonstrates that SMVT, responsible for biotin uptake is functionally active in PC-3 cells.
Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, 2464 Charlotte Street, Kansas City, MO 64108, USA.
Mitesh
M
Patel
Ramya Krishna
RK
Vadlapatla
Sujay
S
Shah
Ashim K
AK
Mitra
D000584:Amiloride / Q000494:pharmacology; D001710:Biotin / Q000378:metabolism*; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000494:pharmacology; D062686:Epithelial Sodium Channel Blockers / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D010042:Ouabain / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D027981:Symporters / Q000235:genetics* / Q000378:metabolism*; D014815:Vitamins / Q000494:pharmacology
D004791:Enzyme Inhibitors; D062686:Epithelial Sodium Channel Blockers; D012333:RNA, Messenger; D027981:Symporters; D014815:Vitamins; C119981:biotin transporter; D010042:Ouabain; D001710:Biotin; D000584:Amiloride; D000254:Sodium-Potassium-Exchanging ATPase