额外的纺锤体极体如1(ESPL1)与多种癌症的发展有关,包括膀胱癌,并且与化学抗性密切相关。在这项研究中,我们旨在揭示ESPL1在膀胱癌进展和顺铂(DDP)耐药中的作用.首先,发现ESPL1在膀胱癌的肿瘤组织和细胞中高表达,在顺铂耐药的肿瘤组织或细胞中高表达。通过Jaspar数据库预测PAX2在ESPL1启动子区域的结合,并通过Ch-IP分析和荧光素酶报告基因测定进行验证。接下来,建立顺铂耐药的T24细胞(T24/DDP),并用ESPL1siRNA(si-ESPL1)或过表达载体(pcDNA-ESPL1)或与PAX2siRNA(si-PAX2)或过表达载体(pcDNA-PAX2)共转染,然后用DDP或AG490(JAK2的抑制剂)处理。结果表明,沉默ESPL1能显著降低T24/DDP细胞的活力,菌落形成和入侵,增强对DDP的敏感性,诱导细胞凋亡。沉默PAX2降低ESPL1表达,增强对DDP的敏感性,诱导T24/DDP细胞凋亡,并抑制JAK2/STAT3通路的激活。过表达ESPL1逆转了PAX2沉默对T24/DDP细胞的影响,而AG490抵消了过表达ESPL1的逆转效应。最后,建立异种移植肿瘤模型,发现沉默ESPL1或DDP治疗抑制肿瘤生长,而沉默ESPL1联合DDP治疗效果最好。总之,本研究提示PAX2介导的ESPL1转录激活通过激活JAK2/STAT3通路增强膀胱癌顺铂耐药。
Extra spindle-polar body like 1 (ESPL1) is associated with the development of a variety of cancers, including bladder cancer, and is closely related to chemoresistance. In this study, we aimed to reveal the role of ESPL1 in bladder cancer progression and cisplatin (DDP) resistance. First, ESPL1 was found to be highly expressed in tumor tissues and cells of bladder cancer, and more highly expressed in cisplatin resistant tumor tissues or cells. The binding of PAX2 in ESPL1 promoter region was predicted by Jaspar database and verified by Ch-IP analysis and the luciferase reporter gene assay. Next, cisplatin-resistant T24 cells (T24/DDP) were established and transfected with ESPL1 siRNA (si-ESPL1) or overexpression vector (pcDNA-ESPL1) or co-transfected with
PAX2 siRNA (si-
PAX2) or overexpression vector (pcDNA-
PAX2), and then treated with DDP or AG490, an inhibitor of JAK2. The results showed that silencing ESPL1 significantly reduced T24/DDP cell viability, colony formation and invasion, enhanced sensitivity to DDP, and induced cell apoptosis. Silencing
PAX2 decreased ESPL1 expression, enhanced sensitivity to DDP, and induced apoptosis of T24/DDP cells, and inhibited activation of JAK2/STAT3 pathway. Overexpressing ESPL1 reversed the effect of
PAX2 silencing on T24/DDP cells, while AG490 counteracted the reversal effect of overexpressing ESPL1. Finally, a xenograft tumor model was established and found that silencing ESPL1 or DDP treatment inhibited tumor growth, while silencing ESPL1 combined with DDP treatment had the best effect. In summary, this study suggested that
PAX2-mediated ESPL1 transcriptional activation enhanced cisplatin resistance in bladder cancer by activating JAK2/STAT3 pathway.