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目的探讨慢性粒细胞白血病(CML)错配修复(MMR)基因的表达水平及调控机制。方法用半定量RT-PCR方法检测62例CML患者及K562细胞的5个MMR基因(hMSH2、hMSH3、 hMSH6、hMLH1、hPMS2)mRNA的表达;用RT．PCR方法动态检测26例进行异基因外周血造血干细胞移植(allo-PBSCT)及4例使用伊马替尼治疗的CML患者bcr．abl及MMR mRNA的表达水平;用伊马替尼体外作用于CML患者的单个核细胞(MNC)及K562细胞后,用Western blot方法检测BCR-ABL融合蛋白的酪氨酸磷酸化水平,RT-PCR方法检测MMR mRNA表达水平。结果与正常人比较,CML患者及K562细胞的hMSH2、hMSH3、hMLH1 mRNA的表达明显降低(P<0．05);26例行allo-PBSCT及4例使用伊马替尼治疗的CML患者,其hMSH2、hMSH3、hMLH1 mRNA的表达随着bcr-abl mRNA的表达降低而升高;伊马替尼体外作用于CML患者MNC及K562细胞后,其hMSH2、hMSH3、hMLH1 mRNA的表达随着BCR-ABL融合蛋白酪氨酸磷酸化水平的降低而升高。结论CML患者、K562细胞hMSH2、 hMSH3、hMLH1 mRNA比正常人降低,bcr-abl融合基因抑制hMSH2、hMSH3、hMLH1 mRNA的表达。
Objective To investigate the expression of MMR gene and its regulatory mechanism in chronic myeloid leukemia (CML). Methods The mRNA expression of five MMR genes (hMSH2, hMSH3, hMSH6, hMLH1, hPMS2) in 62 CML patients and K562 cells was detected by semi-quantitative RT-PCR. PCR method was used to detect 26 cases of allo-PBSCT and 4 cases of CML patients treated with imatinib. abl and MMR mRNA expression levels; using Imatinib in vitro CML patients with mononuclear cells (MNC) and K562 cells, Western blot was used to detect BCR-ABL fusion protein tyrosine phosphorylation level, RT- PCR method to detect MMR mRNA expression levels. Results Compared with normal controls, the expressions of hMSH2, hMSH3 and hMLH1 mRNA in CML patients and K562 cells were significantly decreased (P <0.05). In 26 CML patients treated with allo-PBSCT and 4 patients treated with imatinib, The expression of hMSH2, hMSH3 and hMLH1 mRNA increased with the decrease of bcr-abl mRNA expression. The expression of hMSH2, hMSH3 and hMLH1 mRNA in MNC and K562 cells in vitro after imatinib treatment with BCR-ABL The level of fusion protein tyrosine phosphorylation is decreased and increased. Conclusion In CML patients, the expression of hMSH2, hMSH3 and hMLH1 mRNA in K562 cells is lower than that in normal people, and the bcr-abl fusion gene inhibits the expression of hMSH2, hMSH3 and hMLH1 mRNA.