College of Life Sciences, South China Agricultural University, Guangzhou, China (Z.W., J.Y., K.L., X.W., Z.Z., M.H.); and Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, South China Agricultural University, Guangzhou, China(M.H.).
College of Life Sciences, South China Agricultural University, Guangzhou, China (Z.W., J.Y., K.L., X.W., Z.Z., M.H.); and Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, South China Agricultural University, Guangzhou, China(M.H.)
Drug Metab Dispos. 2024 Oct 16;52(11):1244-1252. doi: 10.1124/dmd.124.001755.
Organic anion transporting polypeptides (OATP, gene symbol ) are well-recognized key determinants for the absorption, distribution, and excretion of a wide spectrum of endogenous and exogenous compounds including many antineoplastic agents. It was therefore proposed as a potential drug target for cancer therapy. In our previous study, it was found that low-dose X-ray and carbon ion irradiation both upregulated the expression of OATP family member OATP1A2 and in turn, led to a more dramatic killing effect when cancer cells were cotreated with antitumor drugs such as methotrexate. In the present study, the underlying mechanism of the phenomenon was explored in breast cancer cell line MCF-7. It was found that the nonreceptor tyrosine kinase v-YES-1 Yamaguchi sarcoma viral oncogene homolog 1 (YES-1) was temporally coordinated with the change of OATP1A2 after irradiation. The overexpression of YES-1 significantly increased OATP1A2 both at the mRNA and protein level. The signal transducer and activator of transcription 3 (STAT3) pathway is likely the downstream target of YES-1 because phosphorylation and nuclear accumulation of STAT3 were both enhanced after overexpressing YES-1 in MCF-7 cells. Further investigation revealed that there are two possible binding sites of STAT3 localized at the upstream sequence of , the encoding gene of OATP1A2. Electrophoretic mobility shift assay and chromatin immunoprecipitation analysis suggested that these two sites bound to STAT3 specifically and the overexpression of YES-1 significantly increased the association of the transcription factor with the putative binding sites. Finally, inhibition or knockdown of YES-1 attenuated the induction effect of radiation on the expression of OATP1A2. SIGNIFICANCE STATEMENT: The present study found that the effect of X-rays on v-YES-1 Yamaguchi sarcoma viral oncogene homolog 1 (YES-1) and organic anion transporting polypeptides (OATP)1A2 was temporally coordinated. YES-1 phosphorylates and increases the nuclear accumulation of signal transducer and activator of transcription 3, which in turn binds to the upstream regulatory sequences of , the coding gene for OATP1A2. Hence, inhibitors of YES-1 may suppress the radiation induction effect on OATP1A2.
有机阴离子转运多肽 (OATP，基因符号) 是广泛的内源性和外源性化合物（包括许多抗肿瘤药物）吸收、分布和排泄的关键决定因素，因此被认为是癌症治疗的潜在药物靶点。在我们之前的研究中，发现低剂量 X 射线和碳离子照射都上调了 OATP 家族成员 OATP1A2 的表达，当癌细胞与抗肿瘤药物（如甲氨蝶呤）联合治疗时，表达上调导致更明显的杀伤效果。在本研究中，在乳腺癌细胞系 MCF-7 中探索了这种现象的潜在机制。结果发现，非受体酪氨酸激酶 v-YES-1 Yamaguchi 肉瘤病毒癌基因同源物 1 (YES-1) 在照射后与 OATP1A2 的变化时间上协调一致。YES-1 的过表达显著增加了 OATP1A2 的 mRNA 和蛋白水平。信号转导和转录激活因子 3 (STAT3) 通路可能是 YES-1 的下游靶点，因为在 MCF-7 细胞中转染 YES-1 后，STAT3 的磷酸化和核积累都增强了。进一步的研究表明，有两个可能的 STAT3 结合位点位于 OATP1A2 编码基因的上游序列。电泳迁移率变动分析和染色质免疫沉淀分析表明，这两个位点特异性地与 STAT3 结合，并且 YES-1 的过表达显著增加了转录因子与假定结合位点的结合。最后，抑制或敲低 YES-1 减弱了辐射对 OATP1A2 表达的诱导作用。 本研究发现 X 射线对 v-YES-1 Yamaguchi 肉瘤病毒癌基因同源物 1 (YES-1) 和有机阴离子转运多肽 (OATP)1A2 的影响在时间上是协调一致的。YES-1 磷酸化并增加了信号转导和转录激活因子 3 的核积累，后者反过来与 OATP1A2 的编码基因的上游调控序列结合。因此，YES-1 的抑制剂可能抑制 OATP1A2 的辐射诱导作用。
