Gynecology Oncology Section, Department of Obstetrics and Gynecology, Women's Health Institute, Cleveland Clinic, Cleveland, Ohio 44195, USA.
Mol Cancer Ther. 2010 Jun;9(6):1709-18. doi: 10.1158/1535-7163.MCT-09-1024. Epub 2010 Jun 1.
Chemotherapy resistance in ovarian cancer remains an unsolved problem in caring for women with this disease. We now show that ovarian cancer immunoreactive antigen domain containing 1 (OCIAD1) has higher expression in chemoresistant compared with chemosensitive ovarian cancer cell lines. We have designed a novel secondary cell homing assay (SCHA) to test the ability of cells to withstand chemotherapy and form secondary colonies that could form recurrent disease. OCIAD1 upregulated cells had significantly higher secondary colony-forming ability than had OCIAD1 downregulated cells following treatment with paclitaxel. Additionally, 18:1 lysophosphatidic acid (LPA) increases OCIAD1 expression in a time- and dose-dependent manner. LPA stimulates OCIAD1 serine phosphorylation within two hours of stimulation. Transfection of MKK6 increases OCIAD1 expression but nuclear translocation is inhibited. Inhibition of p38 mitogen-activated protein kinase blocks LPA-induced OCIAD1 expression. Cycloheximide treatment of MKK6-transfected cells does not inhibit OCIAD1 expression, suggesting that MKK6 upregulation is not translationally controlled. OCIAD1 downregulation knocks down LPA-induced cell adhesion to collagen I and laminin 10/11 and specifically inhibits cell attachment to alpha2, alpha5, alphaV, and beta1 integrins. Proteomic studies indicate that OCIAD1 is physically attached to alpha actin 4 and beta actin. Thus, OCIAD1 may play a role in cytoskeletal function which can alter sensitivity to paclitaxel. This is the first study to indicate that OCIAD1 is a key player in generating ovarian cancer recurrence; it is functionally controlled by LPA and MKK6 signaling, and inhibition of OCIAD1 could be an important strategy in the management of recurrent ovarian cancer.
卵巢癌的化疗耐药性仍然是治疗这种疾病的妇女面临的一个未解决的问题。我们现在表明,卵巢癌免疫反应性抗原结构域 1(OCIAD1)在耐药性卵巢癌细胞系中比在化疗敏感的卵巢癌细胞系中的表达更高。我们设计了一种新的次级细胞归巢测定法(SCHA)来测试细胞耐受化疗并形成可能形成复发性疾病的次级集落的能力。用紫杉醇处理后,OCIAD1 上调的细胞的次级集落形成能力明显高于 OCIAD1 下调的细胞。此外,18:1 溶血磷脂酸(LPA)以时间和剂量依赖的方式增加 OCIAD1 的表达。LPA 在刺激后两小时内刺激 OCIAD1 丝氨酸磷酸化。MKK6 的转染增加了 OCIAD1 的表达,但核易位被抑制。p38 丝裂原活化蛋白激酶的抑制阻断了 LPA 诱导的 OCIAD1 表达。MKK6 转染细胞的环己亚胺处理不会抑制 OCIAD1 的表达,这表明 MKK6 的上调不是翻译控制的。OCIAD1 的下调敲低了 LPA 诱导的细胞对胶原蛋白 I 和层粘连蛋白 10/11 的粘附,并特异性抑制了细胞对 alpha2、alpha5、alphaV 和 beta1 整合素的附着。蛋白质组学研究表明,OCIAD1 与 alpha 肌动蛋白 4 和 beta 肌动蛋白物理结合。因此,OCIAD1 可能在细胞骨架功能中发挥作用,从而改变对紫杉醇的敏感性。这是第一项表明 OCIAD1 是产生卵巢癌复发的关键因素的研究;它受 LPA 和 MKK6 信号的功能控制,抑制 OCIAD1 可能是管理复发性卵巢癌的重要策略。