a-modified-vitro-invasion-assay-to-determine-potential-role-hormones

A Modified <em>In vitro</em> Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the ...

D'Youville University

Murine erythroblasts infected with the anemia-inducing strain of Friend virus (FVA cells) terminally differentiate to the reticulocyte stage after 48 hours of culture in vitro in response to erythropoietin (EPO). The objective of this study was to determine the possible role of proteasome-mediated proteolysis during the terminal differentiation of FVA cells.

The effect of proteasome inhibitors on mammalian erythroid terminal differentiation.

&#x65B0;&#x9C9C;&#x4EBA;&#x7C7B;&#x5B50;&#x5BAB;&#x5185;&#x819C;&#x5916;&#x690D;&#x4F53;&#x88AB;&#x5B75;&#x5316;&#x4E3A; 24 h &#x5728;&#x4E0E;&#x4E09;&#x82EF;&#x6C27;&#x80FA; (10-100 &#x5FAE; M) &#x6216;&#x8F66;&#x8F86; (0.1%&#x4E59;&#x9187;) 37 &#x6444;&#x6C0F;&#x5EA6;&#x3002;&#x6587;&#x5316;&#x5A92;&#x4F53;&#x4E2D;&#x786E;&#x5B9A;&#x4E86;&#x4E09;&#x4E2A;&#x5373; &alpha;-&#x7F9F;&#x57FA;&#x4ED6;&#x83AB;&#x6614;&#x82AC;&#x4EE3;&#x8C22;&#x3001; 4-&#x7F9F;&#x57FA;&#x4ED6;&#x83AB;&#x6614;&#x82AC;&#x548C; N desmethyltamoxifen&#x3002;&#x7EC4;&#x7EC7;&#x5927;&#x5C0F;&#x6709;&#x9650;&#x4F46; DNA &#x52A0;&#x5408;&#x7269;&#x5F62;&#x6210;&#x7684; &alpha;-&#x7F9F;&#x57FA;&#x4ED6;&#x83AB;&#x6614;&#x82AC;&#x901A;&#x8DEF;&#x88AB;&#x68C0;&#x6D4B;&#x5230;&#x4F7F;&#x7528;&#x6B63;&#x5B97; alpha-(deoxyguanosyl-N(2)) &#x4E09;&#x82EF;&#x6C27;&#x80FA;&#x6807;&#x51C6;&#x3002;&#x5206;&#x522B;&#x540E;&#x5B75;&#x5316;&#x4E0E; 100&#x3001; 25&#x3001; 10 &#x5FAE;&#x578B; M &#x4E09;&#x82EF;&#x6C27;&#x80FA;&#xFF0C;&#x68C0;&#x6D4B;&#x5230; 2.45&#x3001; 1.12 &#x548C; 0.44 &#x6BCF; 10(6) &#x6838;&#x82F7;&#x9178;&#x7684; DNA &#x52A0;&#x5408;&#x7269;&#x7684;&#x76F8;&#x5BF9;&#x6C34;&#x5E73;&#x3002;&#x5916;&#x690D;&#x4F53;&#x5E76;&#x53D1;&#x66B4;&#x9732;&#x4E8E; 1 &#x6BEB;&#x7C73;&#x6297;&#x574F;&#x8840;&#x9178;&#x4E0E;&#x4E09;&#x82EF;&#x6C27;&#x80FA;&#x5FAE; M 100 &#x5927;&#x5E45;&#x5EA6;&#x51CF;&#x5C11;&#xFF0C;&alpha;-&#x7F9F;&#x57FA;&#x4ED6;&#x83AB;&#x6614;&#x82AC;&#x7684;&#x6C34;&#x5E73; (68.9%)&#x3002;&#x5F62;&#x6210;&#x7684;&#x4E09;&#x82EF;&#x6C27;&#x80FA;-DNA &#x52A0;&#x5408;&#x7269;&#x63A2;&#x6D4B;&#x5728;&#x5916;&#x690D;&#x4F53;&#x7684;&#x76F8;&#x540C;&#x6807;&#x672C; 100 &#x5FAE; M &#x4E09;&#x82EF;&#x6C27;&#x80FA;&#x4E0E; 1 &#x6BEB;&#x7C73;&#x6297;&#x574F;&#x8840;&#x9178;&#x63A5;&#x89E6;&#x4E5F;&#x88AB;&#x6291;&#x5236;&#x3002;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x652F;&#x6301;&#x7684;&#x4E09;&#x82EF;&#x6C27;&#x80FA;&#x5728;&#x540E;&#x6765;&#x5F62;&#x6210;&#x7684; DNA &#x6C27;&#x5316;&#x751F;&#x7269;&#x8F6C;&#x5316;&#x4F5C;&#x7528;&#x52A0;&#x5408;&#x7269;&#x5728;&#x8FD9;&#x79CD;&#x7EC4;&#x7EC7;&#x3002;

&#x6297;&#x6C27;&#x5316;&#x5242;&#x6291;&#x5236;&#x4E09;&#x82EF;&#x6C27;&#x80FA;-DNA &#x52A0;&#x5408;&#x7269;&#x5728;&#x5B50;&#x5BAB;&#x5185;&#x819C;&#x5916;&#x690D;&#x4F53;&#x57F9;&#x517B;&#x3002;

Fresh human endometrial explants were incubated for 24h at 37 degrees C with either tamoxifen (10-100 micro M) or the vehicle (0.1% ethanol). Three metabolites namely, alpha-hydroxytamoxifen, 4-hydroxytamoxifen, and N-desmethyltamoxifen were identified in the culture media. Tissue size was limited but DNA adducts formed by the alpha-hydroxytamoxifen pathway were detected using authentic alpha-(deoxyguanosyl-N(2)) tamoxifen standards. Relative DNA-adduct levels of 2.45, 1.12, and 0.44 per 10(6) nucleotides were detected following incubations with 100, 25, and 10 micro M tamoxifen, respectively. The concurrent exposure of the explants to 100 micro M tamoxifen with 1mM ascorbic acid reduced the level of alpha-hydroxytamoxifen substantially (68.9%). The formation of tamoxifen-DNA adducts detectable in the explants from the same specimens exposed to 100 micro M tamoxifen with 1mM ascorbic acid were also inhibited. These results support the role of oxidative biotransformation of tamoxifen in the subsequent formation of DNA adducts in this tissue.

Antioxidant inhibits tamoxifen-DNA adducts in endometrial explant culture.

Frais de l&#x26;#39;homme des explants endom&#xE9;triaux ont &#xE9;t&#xE9; incub&#xE9;es pendant 24 heures &#xE0; 37 &#xB0; C avec du tamoxif&#xE8;ne (10-100 micro M) ou le v&#xE9;hicule (0,1% d&#x26;#39;&#xE9;thanol). Trois m&#xE9;tabolites nomm&#xE9;ment l&#x26;#39;alpha-hydroxytamoxif&#xE8;ne, 4-hydroxytamoxif&#xE8;ne, et N-desm&#xE9;thyltamoxif&#xE8;ne ont &#xE9;t&#xE9; identifi&#xE9;es dans les milieux de culture. La taille des tissus a &#xE9;t&#xE9; limit&#xE9;, mais adduits &#xE0; l&#x26;#39;ADN form&#xE9;s par la voie alpha-hydroxytamoxif&#xE8;ne ont &#xE9;t&#xE9; d&#xE9;tect&#xE9;s &#xE0; l&#x26;#39;aide d&#x26;#39;authentiques alpha-(deoxyguanosyl-N (2)) des normes du tamoxif&#xE8;ne. Relatifs adduits &#xE0; l&#x26;#39;ADN des niveaux de 2,45, 1,12, et 0,44 pour 10 (6) nucl&#xE9;otides ont &#xE9;t&#xE9; d&#xE9;tect&#xE9;s &#xE0; la suite des incubations avec 100, 25 et 10 micro M tamoxif&#xE8;ne, respectivement. L&#x26;#39;exposition simultan&#xE9;e des explants &#xE0; 100 M tamoxif&#xE8;ne micro avec l&#x26;#39;acide ascorbique 1 mM r&#xE9;duit le niveau d&#x26;#39;alpha-hydroxytamoxif&#xE8;ne sensiblement (68,9%). La formation du tamoxif&#xE8;ne-adduits ADN d&#xE9;tectables dans les explants des m&#xEA;mes sp&#xE9;cimens expos&#xE9;s &#xE0; 100 M tamoxif&#xE8;ne micro avec l&#x26;#39;acide ascorbique 1 mM ont &#xE9;galement &#xE9;t&#xE9; inhib&#xE9;e. Ces r&#xE9;sultats confirment le r&#xF4;le de la biotransformation oxydative du tamoxif&#xE8;ne dans la formation ult&#xE9;rieure d&#x26;#39;adduits d&#x26;#39;ADN dans ce tissu.

Antioxydant Inhibe tamoxif&#xE8;ne-adduits ADN dans la culture des explants l&#x26;#39;endom&#xE8;tre

Antioxidans hemmt Tamoxifen-DNA-Addukten in endometrialen Explantatkultur.

Espianti dell&#39;endometrio umani freschi sono stati incubati per 24 h a 37 gradi C con tamoxifene (10-100 micro M) o il veicolo (0,1% etanolo). In terreni di coltura sono stati identificati tre metaboliti vale a dire, alfa-hydroxytamoxifen, 4-hydroxytamoxifen e N-desmethyltamoxifen. Tessuto dimensione era limitata ma addotti di DNA formato da via del alfa-hydroxytamoxifen sono stati rilevati utilizzando autentici alpha-(deoxyguanosyl-N(2)) tamoxifene standard. Livelli relativi di addotti del DNA di 2,45 1,12 e 0,44 per 10 nucleotidi sono stati rilevati dopo incubazione con 100, 25 e 10 micro M tamoxifene, rispettivamente. L&#39;esposizione simultanea degli espianti di 100 micro M tamoxifene con acido ascorbico 1 mM ridotto sostanzialmente il livello di alfa-hydroxytamoxifen (68,9%). La formazione di tamoxifen-DNA addotti rilevabile in espianti dallo stesso esemplari esposti al tamoxifene micro M 100 con acido ascorbico 1 mM sono state inoltre inibiti. Questi risultati supporto il ruolo di biotrasformazione ossidativa di tamoxifen la successiva formazione di DNA addotti in questo tessuto.

Antiossidante inibisce il tamoxifene-DNA addotti nella cultura espianto dell&#39;endometrio.

&#x65B0;&#x9BAE;&#x306A;&#x30D2;&#x30C8;&#x5B50;&#x5BAE;&#x5185;&#x819C;&#x306E;&#x5916;&#x690D;&#x7247;&#x306F;&#x3001;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#xFF08;10-100&#x30DE;&#x30A4;&#x30AF;&#x30ED;M&#xFF09;&#x307E;&#x305F;&#x306F;&#x30D3;&#x30D2;&#x30AF;&#x30EB;&#xFF08;0.1&#xFF05;&#x30A8;&#x30BF;&#x30CE;&#x30FC;&#x30EB;&#xFF09;&#x306E;&#x3044;&#x305A;&#x308C;&#x304B;&#x3067;37&#x5EA6;C&#x3067;24&#x6642;&#x9593;&#x30A4;&#x30F3;&#x30AD;&#x30E5;&#x30D9;&#x30FC;&#x30C8;&#x3057;&#x305F;&#x3002;&#x3059;&#x306A;&#x308F;&#x3061;3&#x3064;&#x306E;&#x4EE3;&#x8B1D;&#x7269;&#x3001;&#x3B1;-&#x30D2;&#x30C9;&#x30ED;&#x30AD;&#x30B7;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x3001;4  - &#x30D2;&#x30C9;&#x30ED;&#x30AD;&#x30B7;&#x3001;&#x304A;&#x3088;&#x3073;N-desmethyltamoxifen&#x306F;&#x3001;&#x57F9;&#x990A;&#x57F9;&#x5730;&#x4E2D;&#x3067;&#x540C;&#x5B9A;&#x3055;&#x308C;&#x305F;&#x3002;&#x7D44;&#x7E54;&#x306E;&#x30B5;&#x30A4;&#x30BA;&#x304C;&#x5236;&#x9650;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3057;&#x305F;&#x304C;&#x3001;&#x30A2;&#x30EB;&#x30D5;&#x30A1; - &#x30D2;&#x30C9;&#x30ED;&#x30AD;&#x30B7;&#x7D4C;&#x8DEF;&#x306B;&#x3088;&#x3063;&#x3066;&#x5F62;&#x6210;&#x3055;&#x308C;&#x305F;DNA&#x4ED8;&#x52A0;&#x4F53;&#x306F;&#x672C;&#x7269;&#x306E;&#x3B1;-&#xFF08;deoxyguanosyl-N&#xFF08;2&#xFF09;&#xFF09;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x306E;&#x57FA;&#x6E96;&#x3092;&#x7528;&#x3044;&#x3066;&#x691C;&#x51FA;&#x3057;&#x305F;&#x3002; 2.45&#x3001;1.12&#x3001;10&#x3042;&#x305F;&#x308A;0.44&#x306E;&#x76F8;&#x5BFE;&#x7684;&#x306A;DNA&#x4ED8;&#x52A0;&#x4F53;&#x30EC;&#x30D9;&#x30EB;&#x306F;&#x3001;&#xFF08;6&#xFF09;&#x30CC;&#x30AF;&#x30EC;&#x30AA;&#x30C1;&#x30C9;&#x306F;&#x305D;&#x308C;&#x305E;&#x308C;100&#x3001;25&#x3001;10&#x30DE;&#x30A4;&#x30AF;&#x30ED;M&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x3068;&#x306E;&#x30A4;&#x30F3;&#x30AD;&#x30E5;&#x30D9;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x5F8C;&#x306B;&#x691C;&#x51FA;&#x3055;&#x308C;&#x305F;&#x3002; 1mM&#x306E;&#x30A2;&#x30B9;&#x30B3;&#x30EB;&#x30D3;&#x30F3;&#x9178;&#x3068;100&#x30DE;&#x30A4;&#x30AF;&#x30ED;M&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x306E;&#x5916;&#x690D;&#x7247;&#x306E;&#x540C;&#x6642;&#x66B4;&#x9732;&#x304C;&#xFF08;68.9&#xFF05;&#xFF09;&#x306F;&#x3001;&#x5B9F;&#x8CEA;&#x7684;&#x306B;&#x3B1;-&#x30D2;&#x30C9;&#x30ED;&#x30AD;&#x30B7;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x306E;&#x30EC;&#x30D9;&#x30EB;&#x3092;&#x6E1B;&#x5C11;&#x3055;&#x305B;&#x305F;&#x3002; 1mM&#x306E;&#x30A2;&#x30B9;&#x30B3;&#x30EB;&#x30D3;&#x30F3;&#x9178;&#x3092;100&#x30DE;&#x30A4;&#x30AF;&#x30ED;M&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x306B;&#x3055;&#x3089;&#x3055;&#x308C;&#x3001;&#x540C;&#x3058;&#x691C;&#x4F53;&#x304B;&#x3089;&#x306E;&#x5916;&#x690D;&#x7247;&#x306E;&#x691C;&#x51FA;&#x53EF;&#x80FD;&#x306A;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;-DNA&#x4ED8;&#x52A0;&#x4F53;&#x306E;&#x5F62;&#x6210;&#x3082;&#x6291;&#x5236;&#x3055;&#x308C;&#x305F;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x306F;&#x3001;&#x3053;&#x306E;&#x7D44;&#x7E54;&#x306B;&#x304A;&#x3051;&#x308B;DNA&#x4ED8;&#x52A0;&#x4F53;&#x306E;&#x305D;&#x306E;&#x5F8C;&#x306E;&#x5F62;&#x6210;&#x306B;&#x304A;&#x3051;&#x308B;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;&#x306E;&#x9178;&#x5316;&#x7684;&#x751F;&#x4F53;&#x5185;&#x306E;&#x5F79;&#x5272;&#x3092;&#x30B5;&#x30DD;&#x30FC;&#x30C8;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#x9178;&#x5316;&#x9632;&#x6B62;&#x5264;&#x3068;&#x3057;&#x3066;&#x306F;&#x3001;&#x5B50;&#x5BAE;&#x5185;&#x819C;&#x79FB;&#x690D;&#x7247;&#x57F9;&#x990A;&#x306B;&#x304A;&#x3051;&#x308B;&#x30BF;&#x30E2;&#x30AD;&#x30B7;&#x30D5;&#x30A7;&#x30F3;-DNA&#x4ED8;&#x52A0;&#x4F53;&#x3092;&#x963B;&#x5BB3;&#x3059;&#x308B;

&#xC2E0;&#xC120;&#xD55C; &#xC778;&#xAC04;&#xC758; endometrial explants tamoxifen (10-100 &#xB9C8;&#xC774;&#xD06C;&#xB85C; M) &#xB610;&#xB294; &#xCC28;&#xB7C9; (0.1% &#xC5D0;&#xD0C4;&#xC62C;) 37&#xB3C4; C&#xC5D0;&#xC11C; 24 &#xC2DC;&#xAC04; &#xC778; &#xD050;&#xBCA0;&#xC774; &#xD305; &#xD588;&#xB2E4;. 3 &#xB300;&#xC0AC; &#xC0B0;&#xBB3C; &#xC989;, &#xC54C;&#xD30C; hydroxytamoxifen, 4-hydroxytamoxifen, &#xBC0F; N-desmethyltamoxifen &#xBB38;&#xD654; &#xB9E4;&#xCCB4;&#xC5D0;&#xC11C; &#xD655;&#xC778; &#xB418;&#xC5C8;&#xB2E4;. &#xC870;&#xC9C1; &#xD06C;&#xAE30; &#xC81C;&#xD55C; &#xD558;&#xC9C0;&#xB9CC; DNA &#xC0DD;&#xC131;&#xBB3C; &#xC54C;&#xD30C; hydroxytamoxifen &#xD1B5;&#xB85C; &#xC758;&#xD574; &#xD615;&#xC131; &#xB41C; &#xC815;&#xD1B5; alpha-(deoxyguanosyl-N(2)) tamoxifen &#xD45C;&#xC900;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xAC80;&#xC0C9; &#xB41C;. 2.45, 1.12, &#xC2DC;&#xBBFC;&#xAD8C;&#xC774; &#xB274;&#xD074;&#xB808;&#xC624;&#xD2F0;&#xB4DC; &#xB2F9; 0.44&#xC758; &#xC0C1;&#xB300; DNA adduct &#xC218;&#xC900; &#xB2E4;&#xC74C; 100, 25, 10 &#xB9C8;&#xC774;&#xD06C;&#xB85C; M tamoxifen&#xC640; &#xC678;&#xD53C; &#xAC01;&#xAC01; &#xAC80;&#xCD9C; &#xB410;&#xB2E4;. 1Mm ascorbic &#xC0B0; 100 &#xB9C8;&#xC774;&#xD06C;&#xB85C; M tamoxifen&#xC740; explants&#xC758; &#xB3D9;&#xC2DC; &#xB178;&#xCD9C; &#xC54C;&#xD30C; hydroxytamoxifen &#xC218;&#xC900;&#xC744; &#xC2E4;&#xC9C8;&#xC801;&#xC73C;&#xB85C; &#xAC10;&#xC18C; (68.9%). Tamoxifen DNA &#xD615;&#xC131; &#xC0DD;&#xC131;&#xBB3C; &#xAC19;&#xC740; explants&#xC5D0;&#xC11C; &#xAC10;&#xC9C0; &#xD45C;&#xBCF8;&#xC744; 1mm ascorbic &#xC0B0; 100 &#xB9C8;&#xC774;&#xD06C;&#xB85C; M tamoxifen&#xC5D0; &#xB178;&#xCD9C;&#xB3C4; &#xC800;&#xD574; &#xD588;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xACB0;&#xACFC; &#xC9C0;&#xC6D0; &#xC774;&#xD6C4; DNA &#xD615;&#xC131;&#xC5D0; tamoxifen&#xC758; &#xC0B0;&#xD654; biotransformation &#xC5ED;&#xD560; &#xC0DD;&#xC131;&#xBB3C;&#xC774; &#xC870;&#xC9C1;&#xC5D0;.

&#xD56D; &#xC0B0;&#xD654; &#xC5B5;&#xC81C; tamoxifen DNA endometrial explant &#xBB38;&#xD654;&#xC5D0;&#xC11C; &#xC0DD;&#xC131;&#xBB3C;.

Fresh explantes endometriales humanos se incubaron durante 24 horas a 37 grados C, ya sea con el tamoxifeno (10-100 micro M) o el veh&#xED;culo (0,1% de etanol). Tres metabolitos es decir, alfa-hydroxytamoxifen, 4 hidroxitamoxifeno-, y N-desmetiltamoxifeno fueron identificados en los medios de cultivo. El tejido se tama&#xF1;o limitado, pero aductos de ADN formados por la v&#xED;a de alfa-hidroxitamoxifeno se detectaron utilizando aut&#xE9;nticas alfa-(N-deoxyguanosyl (2)) normas tamoxifeno. Relativos aductos ADN-niveles de 2,45, 1,12, y 0,44 por 10 (6) nucle&#xF3;tidos se detectaron despu&#xE9;s de incubaciones con 100, 25 y 10 micro M tamoxifeno, respectivamente. La exposici&#xF3;n simult&#xE1;nea de los explantes a 100 micro M tamoxifeno con &#xE1;cido asc&#xF3;rbico 1 mM redujo el nivel de alfa-hidroxitamoxifeno considerablemente (68,9%). La formaci&#xF3;n de tamoxifeno aductos de ADN detectables en los explantes procedentes de las mismas muestras expuestas a 100 tamoxifeno micro M con &#xE1;cido asc&#xF3;rbico 1 mM fueron tambi&#xE9;n inhibido. Estos resultados apoyan el papel de la biotransformaci&#xF3;n oxidativa del tamoxifeno en la subsiguiente formaci&#xF3;n de aductos de ADN en este tejido.

Antioxidante inhibe el ADN tamoxifeno-aductos en el explante cultura del endometrio

Although long-term tamoxifen therapy is associated with increased risk of endometrial cancer, little is known about the ability of endometrial tissue to biotransform tamoxifen to potentially reactive intermediates, capable of forming DNA adducts. The present study examined whether explant cultures of human endometrium provide a suitable in vitro model to investigate the tissue-specific biotransformation of tamoxifen. Fresh human endometrial tissue, microscopically uninvolved in disease, was cut into 1 x 2-mm uniform explants and incubated with media containing either 25 or 100 microM tamoxifen in a 24-well plate. Metabolites were analyzed by reversed-phase HPLC using postcolumn, online, photochemical activation and fluorescence detection. Three metabolites, namely, alpha-hydroxytamoxifen, 4-hydroxytamoxifen, and N-desmethyltamoxifen were identified in culture medium and tissue lysates. N-desmethyltamoxifen was found to be the major metabolite in both tissue and media extracts of tamoxifen-exposed explants. Incubations of tamoxifen with recombinant human cytochrome P-450s (CYPs) found that CYP2C9 and CYP2D6 produced all three of the above tamoxifen metabolites, while CYP1A1 and CYP3A4 catalyzed the formation of alpha-hydroxytamoxifen and N-desmethyltamoxifen, and CYP1A2 and CYP1B1 only formed the alpha-hydroxy metabolite. CYP2D6 exhibited the greatest activity for the formation of all three tamoxifen metabolites. Western immunoblots of microsomes from human endometrium detected the presence of CYPs 2C9, 3A, 1A1 and 1B1 in fresh endometrium, while CYPs 2D6 and 1A2 were not detected. Immunohistochemical (IHC) analysis also confirmed the presence of CYPs 2C9, 3A and 1B1 in fresh human endometrium and in viable tissue cultured for 24 h with or without tamoxifen. Together, the results support the use of explant cultures of human endometrium as a suitable in vitro model to investigate the biotransformation of tamoxifen in this target tissue. In addition, the results support the role of CYPs 2C9, 3A, 1A1 and 1B1 in the biotransformation of tamoxifen, including the formation of the DNA reactive alpha-hydroxytamoxifen metabolite, in human endometrium.

Article	Year
The effect of proteasome inhibitors on mammalian erythroid terminal differentiation. Experimental hematology\| PubMed ID: 12135659	2002
Antioxidant inhibits tamoxifen-DNA adducts in endometrial explant culture. Biochemical and biophysical research communications\| PubMed ID: 12849995	2003
Biotransformation of tamoxifen in a human endometrial explant culture model. Chemico-biological interactions\| PubMed ID: 14642736	2003

Diane Bofinger

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