The oncogene Gankyrin is expressed in testicular cancer and contributes to cisplatin sensitivity in embryonal carcinoma cells
The oncogene Gankyrin is expressed in testicular cancer and contributes to cisplatin sensitivity in embryonal carcinoma cells
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DOI：
10.1186/s12885-019-6340-7
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发表时间：
2019-11-19
期刊：
影响因子：
3.8
通讯作者：
Mitchell, Rod T.
中科院分区：
文献类型：
--
作者：
Camacho-Moll, Maria E.;Macdonald, Joni;Mitchell, Rod T.
背景 睾丸生殖细胞癌 (TGCC) 由恶变前原位生殖肿瘤 (GCNIS) 细胞发展而来。 GCNIS起源于胎儿生殖细胞（POU5F1（+）/MAGE-A4（-）），其未能分化为前精原细胞（POU5F1（-）/MAGE-A4（+））并发生恶性转化。 Gankyrin 是一种癌基因，已被证明可以防止 POU5F1 降解，并在肝细胞癌细胞 (HCC) 中与 MAGE-A4 特异性相互作用。我们的目的是研究 Gankyrin 在从生殖细胞进展到侵袭前 GCNIS 以及随后的侵袭性 TGCC 中的作用。方法 我们测定了人胎儿睾丸组织（妊娠周 9-20；n = 38）、生精活跃的成人睾丸组织（n = 9）、生殖细胞成熟延迟的人睾丸组织（n = 4）、侵袭前 GCNIS 患者的睾丸组织（n = 6）和侵袭性 TGCC（包括精原细胞瘤（n = 6）和畸胎瘤（n = 7））中的 Gankyrin 表达。通过 siRNA 敲低 NTera2 细胞（源自胚胎癌）中的 Gankyrin 进行体外功能分析。结果 Gankyrin 的生殖细胞表达仅限于人类胎儿睾丸中的前精原细胞亚群。核 Gankyrin 也在儿童和成人浸润前 TGCC 患者的 GCNIS 细胞以及精原细胞瘤和非精原细胞瘤患者的 GCNIS 细胞中表达。在精原细胞瘤细胞和 NTera2 细胞中观察到细胞质表达。 NTera2 细胞中的 Gankyrin 敲低导致通过 TP53 途径介导的细胞凋亡增加，而 POU5F1 表达不受影响。此外，与单独的顺铂治疗相比，NTera2 细胞中的 Gankyrin 敲低增加了顺铂敏感性，Gankyrin 敲低后细胞死亡增加 (13%, p < 0.05)，可能是通过 BAX 和 FAS 进行的。我们的结果表明，在从生殖细胞到前精原细胞的正常转变过程中，生殖细胞中的 Gankyrin 表达发生变化。此外，Gankyrin 定位的变化与侵袭前 GCNIS 进展为侵袭性 TGCC 相关。此外，我们发现 Gankyrin 参与 NTera2 细胞存活的调节，并且 Gankyrin 表达的减少可以调节顺铂敏感性。结论 这些结果表明，控制 Gankyrin 表达可以减少治疗 TGCC 所需的顺铂剂量，有利于减少化疗的剂量依赖性副作用。需要进一步的研究来评估使用体内模型调节 Gankyrin 对 GCNIS/TGCC 的影响。
Background Testicular germ cell cancer (TGCC) develops from pre-malignant germ neoplasia in situ (GCNIS) cells. GCNIS originates from fetal gonocytes (POU5F1(+)/MAGE-A4(-)), which fail to differentiate to pre-spermatogonia (POU5F1(-)/MAGE-A4(+)) and undergo malignant transformation. Gankyrin is an oncogene which has been shown to prevent POU5F1 degradation and specifically interact with MAGE-A4 in hepatocellular carcinoma (HCC) cells. We aimed to investigate the role of Gankyrin in progression from gonocyte to pre-invasive GCNIS and subsequent invasive TGCC. Methods We determined Gankyrin expression in human fetal testicular tissue (gestational weeks 9-20; n = 38), human adult testicular tissue with active spermatogenesis (n = 9), human testicular tissue with germ cell maturation delay (n = 4), testicular tissue from patients with pre-invasive GCNIS (n = 6), and invasive TGCC including seminoma (n = 6) and teratoma (n = 7). Functional analysis was performed in-vitro by siRNA knock-down of Gankyrin in the NTera2 cells (derived from embryonal carcinoma). Results Germ cell expression of Gankyrin was restricted to a sub-population of prespermatogonia in human fetal testes. Nuclear Gankyrin was also expressed in GCNIS cells of childhood and adult pre-invasive TGCC patients, and in GCNIS from seminoma and non-seminoma patients. Cytoplasmic expression was observed in seminoma tumour cells and NTera2 cells. Gankyrin knock-down in NTera2 cells resulted in an increase in apoptosis mediated via the TP53 pathway, whilst POU5F1 expression was unaffected. Furthermore, Gankyrin knock-down in NTera2 cells increased cisplatin sensitivity with an increase in cell death (13%, p < 0.05) following Gankyrin knock-down, when compared to cisplatin treatment alone, likely via BAX and FAS. Our results demonstrate that Gankyrin expression changes in germ cells during normal transition from gonocyte to prespermatogonia. In addition, changes in Gankyrin localisation are associated with progression of pre-invasive GCNIS to invasive TGCC. Furthermore, we found that Gankyrin is involved in the regulation of NTera2 cell survival and that a reduction in Gankyrin expression can modulate cisplatin sensitivity. Conclusions These results suggest that manipulation of Gankyrin expression may reduce the cisplatin dose required for the treatment of TGCC, with benefits in reducing dose-dependent side effects of chemotherapy. Further studies are required in order to assess the effects of modulating Gankyrin on GCNIS/TGCC using in vivo models.