Lewis(y) antigen-mediated positive feedback loop induces and promotes chemotherapeutic resistance in ovarian cancer
Lewis(y) antigen-mediated positive feedback loop induces and promotes chemotherapeutic resistance in ovarian cancer
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Lewis(y)抗原介导的正反馈环路诱导并促进卵巢癌化疗耐药
DOI：
10.3892/ijo.2018.4496
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发表时间：
2018-10-01
影响因子：
5.2
通讯作者：
Lin, Bei
中科院分区：
文献类型：
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作者：
Liu, Juanjuan;Zheng, Mingjun;Lin, Bei
本研究旨在探讨Lewis(Y)抗原与卵巢癌化疗耐药的关系，并阐明其潜在的分子机制。采用免疫组织化学方法检测卵巢癌化疗耐药组织和细胞中Lewis(Y)的表达。采用逆转录定量聚合酶链式反应(RT-qPCR)检测不同卵巢癌化疗耐药细胞中α-1，2-岩藻糖基转移酶(FUT1)的表达。用基因芯片筛选耐药组和敏感组的差异表达基因，然后用RT-qPCR和Western印迹分析进行验证。我们发现卵巢癌细胞中Lewis(Y)和FUT1的表达随着耐药的诱导而显著增加。卵巢癌耐药组织中Lewis(Y)的阳性表达率和阳性强度也显著高于敏感组。与非耐药细胞系相比，差异表达基因主要集中在与跨膜受体蛋白酪氨酸激酶信号通路和细胞增殖正向调控相关的基因。相互作用网络分析预测了参与调控细胞凋亡过程的基因。RT-qPCR证实膜联蛋白A4(ANXA4)、bcl2相互作用杀伤因子(BIk)、跨膜型4、L 6家族成员4(TM4SF4)和Pleckstrin同源域A家族成员1(PHLDA1)在卵巢癌细胞株中的表达存在高度差异。最后，在耐药细胞中，ANXA4在mRNA和蛋白水平上均有表达增加，并且ANXA4含有Lewis(Y)结构。随着Lewis(Y)表达的增加，Bcl2等抗凋亡蛋白的表达也随之增加。用抗体阻断Lewis(Y)后，相关信号通路和凋亡相关蛋白的表达显著减少。这些发现提供了强有力的证据，证明Lewis(Y)是ANXA4膜蛋白结构的一个组成部分。其过表达可异常激活信号通路，调节多个因子的表达，形成正反馈环，诱导卵巢癌细胞产生化疗耐药，最终促进卵巢癌的进展。
The present study aimed to investigate the association between Lewis(y) antigen and chemoresistance in ovarian cancer and to elucidate the underlying molecular mechanisms. Lewis(y) expression in chemoresistant ovarian cancer tissues and cells was detected by immunohistochemistry. alpha 1,2-fucosyltransferase (FUT1) expression in different ovarian cancer chemotherapy-resistant cells was analyzed by reverse transcription-quantitative PCR (RT-qPCR). Genes differentially expressed in the chemoresistant and sensitive groups were screened using a gene chip followed by validation using RT-qPCR and western blot analysis. We found that Lewis(y) and FUT1 expression in ovarian cancer cells was significantly increased following the induction of drug resistance. The positive expression rate and intensity of Lewis(y) in ovarian cancer chemoresistant tissues were also significantly higher than those in the sensitive group. Compared with the non-resistant cell lines, the differentially expressed genes were mainly enriched in the terms related to the transmembrane receptor protein tyrosine kinase signaling pathway and positive regulation of cell proliferation. Interaction network analysis predicted genes participating in the regulation of apoptotic processes. The highly differential expression of Annexin A4 (ANXA4), BCL2 interacting killer (BIK), transmembrane 4 L six family member 4 (TM4SF4) and pleckstrin homology-like domain family A member 1 (PHLDA1) was validated using RT-qPCR in ovarian cancer cell lines. Finally, ANXA4 expression was increased at both the mRNA and protein level in the drug-resistant cells, and in addition, ANXA4 contained a Lewis(y) structure. The expression of Bcl-2 and other antiapoptotic proteins increased with the increase of Lewis(y) expression. After blocking Lewis(y) using an antibody, the expression of the involved signaling pathway and apoptosis-related proteins decreased significantly. These findings provide strong evidence that Lewis(y) is a component of the structure of the ANXA4 membrane protein. Its overexpression can abnormally activate signaling pathways and regulate the expression of a number of factors, forming a positive feedback loop to induce the chemoresistance of ovarian cancer cells, and ultimately promoting the progression of ovarian cancer.