f45198c14902db7b9e3024a7f81c4aef pmc_30018248.pdf da8c75262f9f68bfb43338bd0eede98fa3d61974 pmc_30018248.pdf 61db4e315c16572341a4f4707e4a898a2c4007b3b47a0f3f666777ee15cbb6e0 pmc_30018248.pdf Title: Generation of Immortalised But Unstable Cells after hTERT Introduction in Telomere-Compromised and p53-Deficient vHMECs Subject: Telomeres, the natural ends of chromosomes, hide the linear telomeric DNA from constitutive exposure to the DNA damage response with a lariat structure or t-loop. Progressive telomere shortening associated with DNA replication in the absence of a compensatory mechanism culminates in t-loop collapse and unmasked telomeres. Dysfunctional telomeres can suppress cancer development by engaging replicative senescence or apoptosis, but they can also promote tumour initiation when cell cycle checkpoints are disabled. In this setting, telomere dysfunction promotes increasing chromosome instability (CIN) through breakage-fusion-bridge cycles. Excessive instability may hamper cell proliferation but might allow for the appearance of some rare advantageous mutations that could be selected and ultimately favour neoplastic progression. With the aim of generating pre-malignant immortalised cells, we ectopically expressed telomerase in telomere-compromised variant human mammary epithelial cells (vHMECs), proficient and deficient for p53, and analysed structural and numerical chromosomal aberrations as well as abnormal nuclear morphologies. Importantly, this study provides evidence that while immortalisation of vHMECs at early stages results in an almost stable karyotype, a transient telomere-dependent CIN period—aggravated by p53 deficiency—and followed by hTERT overexpression serves as a mechanism for the generation of immortal unstable cells which, due to their evolving karyotype, could attain additional promoting properties permissive to malignancy. Keywords: human mammary epithelial cells; telomere dysfunction; chromosome instability; p53; hTERT; cancer Author: Aina Bernal, Elisenda Zafon, Daniel Domínguez, Enric Bertran and Laura Tusell Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.17 CreationDate: Tue Jul 17 13:03:16 2018 CEST ModDate: Tue Jul 17 13:03:16 2018 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 21 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2458315 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SKTRPM+URWPalladioL-Ital Type 1 Custom yes yes no 115 0 BPXBTB+URWPalladioL-Bold Type 1 Custom yes yes no 116 0 OXBKYU+TeXGyreAdventor-Bold Type 1 Custom yes yes no 117 0 WLENKM+VnURWPalladioL-Bold Type 1 Custom yes yes no 118 0 CVJWJT+URWPalladioL-Roma Type 1 Custom yes yes no 119 0 DYVFWR+CMSY10 Type 1 Builtin yes yes no 120 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ESSRQF+VnURWPalladioL-Italic Type 1 Custom yes yes no 568 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-01-19 06:54:13 CET RepresentationInformation: pmc_30018248.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2020-08-03 03:49:12 CEST Size: 2458315 Format: PDF Version: 1.5 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: ぉのぴひはつふっぴどはの〠 Item: Title: げづびふぬぴび〠 Children: Item: Title: ぅびぴちぢぬどびとねづのぴ〠はて〠ぉねねはひぴちぬどびづつ〠ちのつ〠ぎはの〭ぉねねはひぴちぬどびづつ〠ぶえきぅぃび〠ぷどぴと〠いどててづひづのぴ〠ぃづぬぬ〠ぃべっぬづ〠こづぴぴどのでび〠 Item: Title: ごとづ〠ぎづでちぴどぶづ〠ぉねばちっぴ〠はて〠ごづぬはねづひづ〭ぅひはびどはの〠はの〠ぴとづ〠かちひべはぴべばづ〠はて〠ぶえきぅぃび〠ぉび〠ぅのとちのっづつ〠ぢべ〠ごちひでづぴづつ〠ば〵〳〠ぉのちっぴどぶちぴどはの〠 Item: Title: ぁぢびづのっづ〠はて〠ぃぉぎ〠しとづの〠とごぅげご〠ぉび〠ぅっぴはばどっちぬぬべ〠ぅへばひづびびづつ〠どの〠ば〵〳〭ぐひはてどっどづのぴ〠すはふので〠ぶえきぅぃび〠 Item: Title: げづつふっづつ〠あふぴ〠ぐづひびどびぴづのぴ〠ぃぉぎ〠どの〠ば〵〳〭いづてどっどづのぴ〠ぶえきぅぃび〠ぉねねはひぴちぬどびづつ〠ぷどぴと〠とごぅげご〠 Item: Title: いどびっふびびどはの〠 Item: Title: きちぴづひどちぬび〠ちのつ〠きづぴとはつび〠 Children: Item: Title: ぃづぬぬ〠がどのづび〠 Item: Title: がづのぴどぶどひちぬ〠ざづっぴはひび〬〠がづのぴどぶどひふび〠ぐひはつふっぴどはの〠ちのつ〠ごひちのびつふっぴどはの〠 Item: Title: しづびぴづひの〠あぬはぴぴどので〠 Item: Title: いひふで〠ごひづちぴねづのぴび〠 Item: Title: くぢぴちどのどので〠きづぴちばとちびづ〠ぃづぬぬび〠ちのつ〠ぅのつ〭ぴは〭ぅのつ〠うふびどはの〠こっはひどので〠ぃひどぴづひどち〠 Item: Title: ぉの〠こどぴふ〠うぬふはひづびっづのっづ〠えべぢひどつどびちぴどはの〠 Item: Title: いぁぐぉ〠ちのつ〠ごづへちび〠げづつ〭じ〠ぐとちぬぬはどつどの〠こぴちどのどので〠 Item: Title: うぬふはひづびっづのぴ〠きどっひはびっはばべ〠ちのつ〠うぬふはひづびっづのぴ〠ぉねちでづび〠 Item: Title: うぬはぷ〠ぃべぴはねづぴひべ〠 Item: Title: こぴちぴどびぴどっちぬ〠ぁのちぬべびどび〠 Item: Title: げづてづひづのっづび Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: SKTRPM+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 121 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Subtype: Link Rect: 418, 353, 455, 364 Annotation: Subtype: Link Rect: 461, 353, 497, 364 Annotation: Subtype: Link Rect: 318, 327, 354, 338 Annotation: Subtype: Link Rect: 241, 288, 278, 299 Annotation: Subtype: Link Rect: 284, 288, 320, 299 Annotation: Subtype: Link Rect: 167, 263, 205, 273 Annotation: Subtype: Link Rect: 211, 263, 247, 273 Annotation: Subtype: Link Rect: 100, 224, 137, 235 Annotation: Subtype: Link Rect: 143, 224, 179, 235 Annotation: Subtype: Link Rect: 219, 185, 257, 196 Annotation: Subtype: Link Rect: 263, 185, 299, 196 Annotation: Subtype: Link Rect: 399, 160, 437, 170 Annotation: Subtype: Link Rect: 443, 160, 479, 170 Annotation: Subtype: Link Rect: 217, 121, 255, 131 Annotation: Subtype: Link Rect: 261, 121, 297, 131 Annotation: Subtype: Link Rect: 437, 82, 474, 93 Annotation: Subtype: Link Rect: 480, 82, 516, 93 Page: Sequence: 19 Annotations: Annotation: Subtype: Link Rect: 394, 727, 431, 738 Annotation: Subtype: Link Rect: 438, 727, 474, 738 Annotation: Subtype: Link Rect: 393, 701, 430, 712 Annotation: Subtype: Link Rect: 436, 701, 472, 712 Annotation: Subtype: Link Rect: 140, 651, 178, 661 Annotation: Subtype: Link Rect: 184, 651, 220, 661 Annotation: Subtype: Link Rect: 100, 599, 137, 609 Annotation: Subtype: Link Rect: 143, 599, 179, 609 Annotation: Subtype: Link Rect: 343, 572, 380, 583 Annotation: Subtype: Link Rect: 387, 572, 423, 583 Annotation: Subtype: Link Rect: 181, 534, 218, 544 Annotation: Subtype: Link Rect: 224, 534, 261, 544 Annotation: Subtype: Link Rect: 339, 508, 377, 519 Annotation: Subtype: Link Rect: 383, 508, 419, 519 Annotation: Subtype: Link Rect: 184, 483, 221, 493 Annotation: Subtype: Link Rect: 227, 483, 263, 493 Annotation: Subtype: Link Rect: 414, 456, 452, 467 Annotation: Subtype: Link Rect: 458, 456, 494, 467 Annotation: Subtype: Link Rect: 100, 418, 137, 428 Annotation: Subtype: Link Rect: 143, 418, 179, 428 Annotation: Subtype: Link Rect: 176, 379, 214, 390 Annotation: Subtype: Link Rect: 220, 379, 256, 390 Annotation: Subtype: Link Rect: 204, 354, 242, 364 Annotation: Subtype: Link Rect: 248, 354, 284, 364 Annotation: Subtype: Link Rect: 100, 315, 137, 325 Annotation: Subtype: Link Rect: 143, 315, 179, 325 Annotation: Subtype: Link Rect: 479, 275, 516, 286 Annotation: Subtype: Link Rect: 100, 263, 136, 273 Annotation: Subtype: Link Rect: 478, 236, 516, 247 Annotation: Subtype: Link Rect: 100, 224, 136, 235 Annotation: Subtype: Link Rect: 478, 198, 515, 209 Annotation: Subtype: Link Rect: 329, 160, 366, 170 Annotation: Subtype: Link Rect: 373, 160, 409, 170 Annotation: Subtype: Link Rect: 305, 133, 342, 144 Annotation: Subtype: Link Rect: 349, 133, 385, 144 Annotation: Subtype: Link Rect: 424, 82, 462, 93 Annotation: Subtype: Link Rect: 468, 82, 504, 93 Page: Sequence: 20 Annotations: Annotation: Subtype: Link Rect: 100, 715, 136, 725 Annotation: Subtype: Link Rect: 422, 688, 459, 699 Annotation: Subtype: Link Rect: 465, 688, 501, 699 Annotation: Subtype: Link Rect: 339, 663, 377, 674 Annotation: Subtype: Link Rect: 383, 663, 419, 674 Annotation: Subtype: Link Rect: 363, 637, 400, 648 Annotation: Subtype: Link Rect: 406, 637, 442, 648 Annotation: Subtype: Link Rect: 158, 599, 194, 609 Annotation: Subtype: Link Rect: 149, 573, 186, 583 Annotation: Subtype: Link Rect: 193, 573, 229, 583 Annotation: Subtype: Link Rect: 469, 533, 505, 546 Annotation: Subtype: Link Rect: 435, 508, 473, 519 Annotation: Subtype: Link Rect: 479, 508, 515, 519 Annotation: Subtype: Link Rect: 246, 469, 284, 480 Annotation: Subtype: Link Rect: 290, 469, 326, 480 Annotation: Subtype: Link Rect: 327, 430, 364, 441 Annotation: Subtype: Link Rect: 371, 430, 407, 441 Annotation: Subtype: Link Rect: 425, 404, 462, 415 Annotation: Subtype: Link Rect: 468, 404, 504, 415 Annotation: Subtype: Link Rect: 378, 378, 416, 390 Annotation: Subtype: Link Rect: 422, 378, 458, 390 Annotation: Subtype: Link Rect: 478, 353, 516, 364 Annotation: Subtype: Link Rect: 100, 341, 136, 351 Annotation: Subtype: Link Rect: 480, 314, 516, 325 Annotation: Subtype: Link Rect: 100, 302, 136, 312 Annotation: Subtype: Link Rect: 149, 263, 187, 273 Annotation: Subtype: Link Rect: 193, 263, 229, 273 Annotation: Subtype: Link Rect: 331, 224, 369, 235 Annotation: Subtype: Link Rect: 375, 224, 411, 235 Annotation: Subtype: Link Rect: 272, 199, 310, 209 Annotation: Subtype: Link Rect: 316, 199, 352, 209 Annotation: Subtype: Link Rect: 438, 159, 475, 170 Annotation: Subtype: Link Rect: 481, 159, 516, 170 Annotation: Subtype: Link Rect: 407, 133, 443, 144 Annotation: Subtype: Link Rect: 246, 107, 282, 118 Annotation: Subtype: Link Rect: 158, 69, 196, 80 Annotation: Subtype: Link Rect: 202, 69, 238, 80 Page: Sequence: 21 Annotations: Annotation: Subtype: Link Rect: 149, 715, 186, 725 Annotation: Subtype: Link Rect: 298, 688, 336, 699 Annotation: Subtype: Link Rect: 331, 663, 368, 674 Annotation: Subtype: Link Rect: 181, 638, 218, 648 Annotation: Subtype: Link Rect: 224, 638, 261, 648 Annotation: Subtype: Link Rect: 382, 611, 419, 622 Annotation: Subtype: Link Rect: 426, 611, 462, 622 Annotation: Subtype: Link Rect: 149, 560, 187, 570 Annotation: Subtype: Link Rect: 193, 560, 229, 570 Annotation: Subtype: Link Rect: 230, 521, 268, 532 Annotation: Subtype: Link Rect: 274, 521, 310, 532 Annotation: Subtype: Link Rect: 478, 482, 516, 493 Annotation: Subtype: Link Rect: 100, 470, 136, 480 Annotation: Subtype: Link Rect: 430, 443, 468, 454 Annotation: Subtype: Link Rect: 474, 443, 510, 454 Annotation: Subtype: Link Rect: 480, 404, 516, 415 Annotation: Subtype: Link Rect: 100, 392, 136, 402 Annotation: Subtype: Link Rect: 76, 350, 166, 381 Annotation: Subtype: Link Rect: 234, 344, 424, 355 Checksum: 7224e6a2 Type: CRC32 Checksum: f45198c14902db7b9e3024a7f81c4aef Type: MD5 Checksum: da8c75262f9f68bfb43338bd0eede98fa3d61974 Type: SHA-1 Checksum: 61db4e315c16572341a4f4707e4a898a2c4007b3b47a0f3f666777ee15cbb6e0 Type: SHA-256