TCRP1在慢性髓系白血病细胞中的表达及其对细胞增殖的影响

The expression of TCRP1 in chronic myeloid leukemia cells and its effects on cell proliferation

目的探讨舌癌耐药相关基因（TCRP1）在慢性髓系白血病细胞增殖中的可能作用，为探寻白血病的致病机制提供新思路。方法用实时定量PCR和Western blot的方法检测慢性髓系白血病患者外周血单核细胞（PBMC）和K562细胞系中TCRP1的表达，在K562细胞中干扰TCRP1的表达，用四氮唑蓝化合物（MTS）和软琼脂克隆形成实验来检测细胞的增殖，用Westernblot实验来检测蛋白激酶B（AKT）的表达及其磷酸化水平。结果在慢性髓系白血病患者PBMC中，TCRP1的mRNA和蛋白水平的表达量均高于正常对照（P〈0．05），MTS实验和软琼脂克隆形成实验结果表明干扰TCRP1表达后，K562细胞的增殖显著减慢（P〈0．05）。干扰TCRP1后，AKT蛋白的表达量不变，而其磷酸化水平显著降低（P〈0．05）。结论TCRP1在慢性髓系白血病细胞中高表达，TCRP1可能通过AKT的磷酸化水平来影响K562细胞的增殖。

Objective To investigate the effects of tongue cancer resistance-associated protein 1 （ TCRP1 ） in proliferation of chronic myeloid leukemia cells （ CML）, and explore the new thoughts of patho- genesis of CML. Methods The expression of TCRP1 was detected in the peripheral blood mononuclear cells （PBMC） of CML with real-time quantitative polymerase chain reaction （PCR） and Western blot. After the expression of TCRP1 was interfered in K562 ceils, the proliferation of cells was detected by 3-（4,5- dimenthyhhiazol-2-yl） -5-（ 3-carboxymethoxyphenyl ） -2-（ 4-sulfophenyl ） -2H-tetrazolium （ MTS ） assay and soft agar colony forming assay, and the expression of protein kinase B （AKT） and its phosphorylation were tested by Western blot. Results In PBMC of CML patients, the mRNA and protein levels of TCRP1 were significantly higher than those of normal controls. The results of MTS assay and soft agar colony forming as- say showed that the proliferation of K562 cells was significantly decreased after the expression of TCRP1 was interfered. After knockdown of TCRP1 in K562 cells, the phosphorylation of AKT was significantly de- creased while the expression of total AKT did not change. Conclusions The expression of TCRP1 was in- creased in CML ceils. High expression of TCRP1 might contribute to proliferation of K562 cells via the phosphorylation of AKT.