Establishing a papillary craniopharyngioma cell line by SV40LT-mediated immortalization
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Establishing a papillary craniopharyngioma cell line by SV40LT‑mediated immortalization Yi Liu1 · Chao‑hu Wang1 · Jun Fan1 · Jun‑xiang Peng1 · Jun Pan1 · Xi’an Zhang1 · Song‑tao Qi1 Accepted: 5 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Background Craniopharyngioma represents a troublesome tumor of the intracranial sellar region. There are currently no available well-characterized craniopharyngioma cell lines. This lack of reliable, immortal cell lines is a major reason for the slow progress in fundamental research related to craniopharyngioma. Methods We describe the development of an immortal papillary craniopharyngioma (PCP) cell line by transfecting primary PCP cells with the pLenti-simian virus 40 large T antigen(SV40LT). Results Three clones have been cultured for more than 14 months so far, while non-transfected cells ceased proliferation within three months of isolation. The established immortal PCP cell lines were identified to have BRAFV600E mutations, while no mutations in tumor suppressor genes were found in primary cells or immortal cells. Immortal cells had higher proliferation rates and formed tumors when implanted in the bran of nude mice. BRAF inhibition in immortal PCP cells altered cell morphology, inhibited cell proliferation and promoted apoptosis. Conclusion We successfully developed PCP cell lines by SV40LT-mediated immortalization. These cell lines represent a powerful tool for fundamental and therapeutical studies on craniopharyngioma. Keywords Papillary craniopharyngioma (PCP) · SV40LT · Immortalization Abbreviations CP Craniopharyngioma aCP Adamantinomatous craniopharyngioma pCP Papillary craniopharyngioma HE Hematoxylin and eosin
Introduction Craniopharyngioma (CP) is an epithelium-derived neoplasm that develops in the sellar region. Two clinically and pathologically distinct CP subtypes have been identified by the WHO [1] adamantinomatous CP (ACP) and papillary CP (PCP). Although CP is almost always benign, they are likely to spread in the hypothalamic-pituitary axis and surrounding Yi Liu, Chao-hu Wang and Jun Fan have contributed equally to this study. * Song‑tao Qi [email protected] 1
Department of Neurosurgery, Nanfang Hospital of Southern Medical University, Guangzhou 510515, China
structures, which may lead to severe postoperative complications after resection [2–4]. The local invasive growth of craniopharyngioma is the leading cause of severe postoperative complications and postoperative recurrence. In the past few years, the research on CP biology has focused on cell proliferation and apoptosis, extracellular matrix organization/degradation, as well as oncogenes and tumor suppressor genes involved [5–11]. However, the molecular mechanisms underlying CP local invasion and recurrence has not yet been fully clarified. The lack of stable and immortal CP cell lines is the most important restraint. Similarly to other benign tumor cells, the survival time of primary CP cells in vitro is generally not more than 2–3 mon
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