Antiproliferative effects of Src inhibition on medullary thyroid cancer
Antiproliferative effects of Src inhibition on medullary thyroid cancer
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DOI：
10.1210/jc.2003-031917
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发表时间：
2004-07-01
影响因子：
5.8
通讯作者：
Nwariaku, FE
中科院分区：
文献类型：
--
作者：
Liu, ZJ;Falola, J;Nwariaku, FE
对于复发性或转移性甲状腺髓样癌（MTC）没有有效的治疗方法。遗传性MTC与RET原癌基因突变有关，RET原癌基因编码酪氨酸激酶。我们假设Src酪氨酸激酶调节MTC的增殖。在Src特异性酪氨酸激酶抑制剂PP2或染料木黄酮的存在下，检测了人MTC细胞系TT的增殖。用Coulter计数器或通过流式细胞术进行细胞计数。通过溴脱氧尿苷掺入法评价DNA合成。使用细胞死亡ELISA来评估细胞凋亡。通过Western免疫印迹测定Akt磷酸化。使用免疫沉淀激酶测定法测量MAPK活性，并且用SB 202190（p38 MAPK）和PD 098059（MAPK激酶）实现MAPK抑制。与对照组相比，PP2降低DNA合成，抑制Akt磷酸化，增加细胞凋亡。MAPK激酶抑制剂，PD098059，衰减DNA合成，而金雀异黄素引起细胞计数和DNA合成和最小的变化在apoptosis.We的结论是Src依赖的MTC增殖发生通过增加DNA合成和减少凋亡的适度下降。后一种效应可能是由Akt存活信号介导的。Src活性的调节是MTC的潜在治疗靶点。
There is no effective treatment for recurrent or metastatic medullary thyroid cancer (MTC). Hereditary MTC is associated with mutations in the RET protooncogene, which encodes for a tyrosine kinase. We postulated that Src tyrosine kinases regulate MTC proliferation.Proliferation of the human MTC cell line, TT, was examined in the presence of a Src-specific tyrosine kinase inhibitor, PP2, or genistein. Cell counts were performed with a Coulter counter or by flow cytometry. DNA synthesis was evaluated by bromodeoxyuridine incorporation. A cell death ELISA was used to assess apoptosis. Akt phosphorylation was determined by Western immunoblot. MAPK activity was measured using an immunoprecipitation kinase assay, and MAPK inhibition was achieved with SB202190 (p38 MAPK) and PD098059 ( MAPK kinase). Data were analyzed by ANOVA.Compared with controls, PP2 reduced DNA synthesis, abolished Akt phosphorylation, and increased apoptosis. The MAPK kinase inhibitor, PD098059, attenuated DNA synthesis, whereas genistein caused modest declines in cell count and DNA synthesis and minimal changes in apoptosis.We conclude that Src-dependent MTC proliferation occurs via increased DNA synthesis and reduced apoptosis. The latter effect may be mediated by Akt survival signals. Modulation of Src activity is a potential therapeutic target in MTC.