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ID naloge: 34 Letnik: 1998 Predmet: genetika
Ugotavljanje genskih sprememb pri raku želodca
Avtor: Andreja Gojkovic, Mojca Grošelj
Mentor: prof. dr. Radovan Komel
IZHODIŠCA. Žlezni rak (adenokarcinom) želodca je pogosto maligno obolenje v Sloveniji. Po Laurenovi razvrstitvi ga delimo na intestinalno, dobro diferencirano, in difuzno, slabo diferencirano obliko. Poti nastanka obeh oblik naj bi se razlikovali po znacilnem zaporedju genetskih sprememb. V naši raziskavi, ki je prva tovrstna v Sloveniji, smo želeli na izbranem vzorcu intestinalne in difuzne oblike adenokarcinomov želodca ugotoviti izgubo alela nekaterih zaviralnih genov (APC, p53, Rb, nm23 in DCC) in ugotoviti morebitno razliko med obema oblikama. Hkrati smo želeli uvesti novo, najsodobnejšo metodo preiskovanja izgube alelov.
BOLNIKI IN METODE. Proucevali smo vzorce tumorskega in zdravega tkiva želodca, odvzete sedemnajstim nakljucno izbranim bolnikom z rakom želodca, operiranim v letu 1997. Za pomnoževanje mikrosatelitnih zaporedij zaviralnih genov smo uporabili verižno reakcijo s polimerazo. Dobljene pridelke smo analizirali po nacelu kapilarne elektroforeze s pomocjo avtomatskega genskega analizatorja. Po izvedbi kapilarne elektroforeze smo izlocili pet parov vzorcev, ker niso bili primerni za nadaljnjo analizo.
REZULTATI. Za gen APC smo ugotovili izgubo heterozigotnosti v 17 %, za gen p53 v 50 % in za gen DCC v 100 % primerov. Pri genih Rb in nm23 nismo ugotovili izgube heterozigotnosti. Vecino izgub heterozigotnosti smo ugotovili pri intestinalni obliki, eno pri mešani obliki in nobene pri difuzni obliki.
ZAKLJUCKI. Uvedli smo najsodobnejšo metodo preiskovanja izgube alelov. Rezultati naše raziskave so za gena APC in p53 primerljivi z rezultati raziskav, opisanih v literaturi, ne pa tudi za gene DCC, nm23 in Rb. Ugotovili smo razliko v pojavljanju genskih sprememb pri intestinalni in difuzni obliki. Naša raziskava je dala osnove za nadaljnje iskanje najugodnejših pogojev za izvedbo obsežnejše multipleks verižne reakcije s polimerazo in za njeno uporabo pri preiskavi še neraziskanih genov.
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[Abstract / English version]
Ugotavljanje genskih sprememb pri raku želodca
Author: Andreja Gojkovic, Mojca Grošelj
Mentor: prof. dr. Radovan Komel
BACKGROUND. Stomach cancer (gastric adenocarcinoma) is a common malignant disease in Slovenia. According to Lauren?s classification we distinguish between well differentiated intestinal and less differentiated diffuse type of the disease. Both types differ in the way they develop, as their sequences of genetic changes are not the same. In the present research, performed for the first time and in selected sample of Slovenian patients, we wanted to investigate allelic loss at several tumor suppressor genes (APC, p53, Rb, nm23 and DCC) and thus distinguish between the intestinal and diffuse type of gastric adenocarcinoma in our population. The aim of this work was also to introduce new and the most recent method for investigation of allelic loss.
PATIENTS AND METHODS. Seventeen randomly selected frozen samples of adenocarcinomas and of surrounding non-affected tissues were obtained from 17 patients surgically treated in 1997. Polymerase chain reaction was used for multiplying microsatellite sequences related to tumor suppressor genes. PCR products were analysed with capillary electrophoresis in automatic gene analyser. After capillary electrophoresis, five sample pairs were excluded from further analysis because of their non-informativity which was due to non-efficient amplification.
RESULTS. Loss of heterozigosity was found in 17% of samples for APC, in 50% for p53, and in 100% for DCC. There were no loss of heterozigosity for Rb and nm23. The major part of loss of heterozigosity was found in intestinal, one in mixed, and none in diffuse type of adenocarcinoma.
CONCLUSIONS. We introduced the most recent method for investigating allelic loss. The results of our investigation correlate with other reports in literature for APC and p53, but not for DCC, nm23 and Rb. We found differences in genetic alterations between intestinal and diffuse type of adenocarcinoma in our population. The present results represent good starting point to look for optimal conditions for introducing multiplex polymerase chain reaction with even larger set of genetic markers and to use it in research of some other, not yet investigated tumor suppressor and mismatch repair genes.
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