Abstract：
Choriocarcinoma is one of the most aggressive gestational trophoblastic neoplasias (GTN). Methotrexate (MTX) resistance is the main cause of treatment failure in choriocarcinoma. However, the mechanism of MTX resistance in choriocarcinoma is poorly known. This study aims to explore the function of Lectin galactoside-binding soluble 3 binding protein (LGALS3BP) in MTX-resistance in choriocarcinoma cells. Gradual dose escalation of MTX was used to establish MTX-resistant choriocarcinoma cells (JAR-MTX and JEG3-MTX cell lines). RNA-sequencing was used to explore the differentially expressed genes. Plasmids or SiRNA transfection was used to regulate the expression of LGALS3BP. ELISA was used to detect the concentrations of LGALS3BP in the serum of MTX-sensitive and MTX-resistant patients. qRT-PCR, Western blot, and CCK-8 assay were used to determine the effects of LGALS3BP on MTX-resistance in JAR and JEG3 cells. The results showed the relative resistance index (RI) of MTX is 791.50 and 1040.04 in JAR-MTX and JEG3-MTX, respectively. LGALS3BP was up-regulated in MTX-resistant cells compared to original cells in both RNA and protein level. The concentrations of LGALS3BP were higher in the sera of MTX-resistant patients than in MTX-sensitive patients. Knocking down LGALS3BP can reverse the MTX-resistance in JAR-MTX and JEG3-MTX cells. In summary, we preliminarily established two MTX-resistant cells, and performed RNA-sequencing, and found LGALS3BP may play important role in MTX-resistance. Our work not only provides a research tool (MTX-resistant cells) for other researchers, but gives some hint on how MTX resistance is regulated.
摘要：
绒毛膜癌是最具侵袭性的妊娠滋养细胞肿瘤(GTN)之一。甲氨蝶呤（MTX）耐药是绒毛膜癌治疗失败的主要原因。然而,绒毛膜癌中MTX耐药的机制尚不清楚。本研究旨在探讨凝集素半乳糖苷结合可溶性3结合蛋白(LGALS3BP)在绒毛膜癌细胞MTX耐药中的作用。使用MTX的逐渐剂量递增来建立MTX抗性绒毛膜癌细胞(JAR-MTX和JEG3-MTX细胞系)。RNA测序用于探索差异表达的基因。使用质粒或SiRNA转染来调节LGALS3BP的表达。采用ELISA法检测MTX敏感和MTX耐药患者血清中LGALS3BP的浓度。qRT-PCR,蛋白质印迹,和CCK-8测定用于确定LGALS3BP对JAR和JEG3细胞中MTX抗性的影响。结果表明，在JAR-MTX和JEG3-MTX中，MTX的相对电阻指数（RI）分别为791.50和1040.04，分别。在RNA和蛋白质水平上,与原始细胞相比,LGALS3BP在MTX抗性细胞中上调。MTX耐药患者血清中LGALS3BP的浓度高于MTX敏感患者。敲除LGALS3BP可以逆转JAR-MTX和JEG3-MTX细胞中的MTX抗性。总之,我们初步建立了两种MTX耐药细胞，进行RNA测序，发现LGALS3BP可能在MTX耐药中起重要作用。我们的工作不仅为其他研究人员提供了研究工具（抗MTX细胞），但给出了一些关于MTX抗性如何调节的提示。
