31 KB143 KB28 KB200520252009Bilač-Krašnja, MojcaČerne, Darkoštevilka:16 straniletnik:60str. 21-26ISSN:0014-8229COBISSID:2540401URN:URN:NBN:SI:doc-8SPJP0WAslSlovensko farmacevtsko društvoFarmacevtski vestnikbiokemijalipoproteinska lipazarak (medicina)Optimizacija analiznega postopka merjenja aktivnosti lipoproteinske lipaze v rakavem tkivu| Optimization of an analytical procedure of lipoprotein lipase activity measurement in cancer tissue|Several studies demonstrated that there is a connection between lipoprotein lipase activity (LPL; EC 3.1.1.34) and cancer tissue growth. An intensive investigation of the role of LPL in cancer progress is limited because of the lack of simple methods for LPL activity measurements. Our aim was to optimize the method which is based on the release of LPL from tissue with the solution containing heparin and albumin, and which is based on enzyme activity measurement with commercially available LPL Activity Kit (Roar Biomedical, NewYork, ZDA). We also wanted to find out if this method is suitable for LPL activity measurement in cancer tissue. We came to the conclusion that the method is reliable only by considering additional - in our study established recommendations regarding the stability of all solutions we used, measurement fluorescent product in 5 minutes intervals, individual estimation of the appropriateness of enzyme reaction as well as standardization enzyme activity on weight of tissue (nkat/kg). Using this method we found out that LPL activity was higher in cancer tissue (1.9-fold median difference) than when compared to adjacent, apparently healthy lung tissue of the same patient (p < 0,001; 10 patients with non-small cell lung cancer without metastases). We conclude that this method is suitable for LPL activity measurement in tissues as well as for further, better understanding of the role of LPL in possible mechanisms for increasing the supply of lipid nutrients to the tumour, necessary for tumour growthRaziskave kažejo na povezavo med aktivnostjo lipoproteinske lipaze (LPL; EC 3.1.1.34) in rastjo rakavega tkiva. Zaradi odsotnosti enostavnih metod za merjenje njene aktivnosti je intenzivnejše proučevanje vloge LPL v razvoju raka omejeno. V raziskavi smo želeli ugotoviti, ali je postopek temelječe na izluževanju LPL iz tkiva z raztopino heparina in albumina ter merjenju aktivnosti encima v eluatu s komercialnim postopkom LPL Activity Kit (Roar Biomedical, New York, ZDA) primeren za merjenje aktivnosti LPL v rakavem tkivu. Ugotovili smo, da je postopek zanesljiv le ob upoštevanju dodatnih, v naši raziskavi ugotovljenih priporočil glede stabilnosti izluževalne raztopine in substratne emulzije, merjenja v encimski reakciji nastalega fluorescenoenega produkta vsakih pet minut, individualne ocene ustreznosti poteka encimske reakcije in standardizacije aktivnosti encima na maso tkiva (nkat/kg). Z vpeljanim postopkom smo ugotovili, da je aktivnost LPL v rakavem pljučnem tkivu v povprečju 1,94-krat višja kot v sosednjem, navidezno neprizadetem pljučnem tkivu istega bolnika (p < 0,001; 10 bolnikov z nedrobnoceliočnim pljučnim rakom brez metastaz). Zaključujemo lahko, da je vpeljan postopek primeren za merjenje aktivnosti LPL v tkivih in za nadaljnje proučevanje vloge encima pri preskrbi rakavega tkiva z maščobami, nujnimi za rast tkivaTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaSlovensko farmacevtsko društvo