Quantification of O⁶-methylguanine-DNA methyltransferase mRNA in human brain tumors

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• 作者：Mineura ; Katsuyoshi ; Watanabe ; Katsuo ; Yanagisawa ; Toshiharu ; Kowada ; Masayoshi
• 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
• 出版年：1996
• 出版时间：February 9, 1996
• 年：1996
• 卷：1289
• 期：1
• 页码：105-109
• 全文大小：414 K

文摘

O⁶-Methylguanine-DNA methyltransferase (MGMT) is strongly involved in drug resistance mechanism of tumor cells to chloroethylnitrosoureas (CENUs), because it removes and repairs CENU-induced O⁶-alkylguanine-DNA by accepting the alkyl group at a cysteine moiety. MGMT activity and MGMT mRNA expression are good indicators for detection of sensitive cells or resistant cells to CENUs. In the present study, we applied a non-radioactive reverse transcription-polymerase chain reaction (RT-PCR) method on quantitative measurement of MGMT mRNA expression. Estimated levels of MGMT mRNA expression determined by this RT-PCR method were consistent with the actual doses of MGMT mRNA. This relationship was noted at a wide range from 10 fg to 10 pg. The relative expression levels of MGMT mRNA estimated from kinetic analysis correlated well with MGMT activity determined using³ H-methylnitrosourea-treated DNA substrate in brain tumor cells (P < 0.001 with a correlation coefficient of 0.997). The RT-PCR method facilitated quantitative measurements in even a small amount of biopsy specimens obtained by stereotactic brain surgery.