{"doi":"10.21203/rs.3.rs-35307/v3","title":"EML4-ALK induces cellular senescence in mortal normal human cells and promotes anchorage-independent growth in hTERT-transduced normal human cells","abstract":"Abstract Background: Chromosomal inversions involving anaplastic lymphoma kinase ( ALK ) and echinoderm microtubule associated protein like 4 ( EML4 ) generate a fusion protein EML4-ALK in non-small cell lung cancer (NSCLC). The understanding of EML4-ALK function can be improved by a functional study using normal human cells. Methods: Here we for the first time conduct such study to examine the effects of EML4-ALK on cell proliferation, cellular senescence, DNA damage, gene expression profiles and transformed phenotypes. Results: The lentiviral expression of EML4-ALK in mortal, normal human fibroblasts caused, through its constitutive ALK kinase activity, an early induction of cellular senescence with accumulated DNA damage, upregulation of p16 INK4A and p21 WAF1 , and senescence-associated b-galactosidase (SA-β-gal) activity. In contrast, when EML4-ALK was expressed in normal human fibroblasts transduced with telomerase reverse transcriptase (hTERT), which is activated in the vast majority of NSCLC, the cells showed accelerated proliferation and acquired anchorage-independent growth ability in soft-agar medium, without accumulated DNA damage, chromosome aberration, nor p53 mutation. EML4-ALK induced the phosphorylation of STAT3 in both mortal and hTERT-transduced cells, but RNA sequencing analysis suggested that the different signaling pathways contributed to the different phenotypic outcomes in these cells. While EML4-ALK also induced anchorage-independent growth in hTERT-immortalized human bronchial epithelial cells in vitro , the expression of EML4-ALK alone did not cause detectable in vivo tumorigenicity in immunodeficient mice. Conclusions: Our data indicate that the expression of hTERT is critical for EML4-ALK to manifest its in vitro transforming activity in human cells. This study provides the isogenic pairs of human cells with and without EML4-ALK expression.","journal":"Research Square","year":2020,"id":133695,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9433,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":589315,"name":"Masaru Matsumoto","orcid":"0000-0001-6647-6624","position":1,"is_corresponding":false},{"id":279503,"name":"Jessica A. Beck","orcid":"0000-0002-0145-7606","position":2,"is_corresponding":false},{"id":286540,"name":"Izumi Horikawa","orcid":"0000-0002-2893-5014","position":3,"is_corresponding":false},{"id":373628,"name":"Takahiro Oike","orcid":"0000-0003-1332-4306","position":4,"is_corresponding":false},{"id":589316,"name":"Hirokazu Okayama","orcid":"0000-0003-0966-4746","position":5,"is_corresponding":false},{"id":314756,"name":"Hiromi Tanaka","orcid":"0000-0002-6348-6058","position":6,"is_corresponding":false},{"id":315253,"name":"Sandra Burkett","orcid":"0000-0002-5476-4028","position":7,"is_corresponding":false},{"id":51783,"name":"Ana I. Robles","orcid":"0000-0001-5019-4374","position":8,"is_corresponding":false},{"id":589317,"name":"Mohammed Khan","orcid":"0009-0000-1202-9829","position":9,"is_corresponding":false},{"id":589318,"name":"Delphine Lissa","orcid":"0000-0002-1684-5134","position":10,"is_corresponding":false},{"id":589319,"name":"Masahiro Seike","orcid":"0000-0002-1572-5472","position":11,"is_corresponding":false},{"id":589320,"name":"Akihiko Gemma","orcid":"0000-0003-0762-2737","position":12,"is_corresponding":false},{"id":53009,"name":"Hiroyuki Mano","orcid":"0000-0003-4645-0181","position":13,"is_corresponding":false},{"id":276012,"name":"Curtis C. Harris","orcid":"0000-0001-5268-0181","position":14,"is_corresponding":false},{"id":589314,"name":"Akihiko Miyanaga","orcid":"0000-0003-2026-9181","position":0,"is_corresponding":true}],"reference_count":38,"raw_metadata":null,"created_at":"2026-07-18T23:16:20.998822Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}