{"doi":"10.33279/jkcd.v10i03.207","title":"ESTABLISHMENT OF MULTIPLE MYELOMA CELL  LINES WITH REDUCED EXPRESSION OF HEPATOCYTE  GROWTH FACTOR USING RNAI APPROACH","abstract":"<jats:p>Objective:\nTo establish the gene knockdown system (RNAi) using different Multiple Myeloma Cell lines (MMCL).\nMaterial and Methods:\nRNAi is generally used in biomedical research to study the outcome of knockdown of gene expression. In this research, retroviral-shRNA expression constructs based on micro RNA-adapted short hairpin RNA (sh RNA mir) were designed and constructed. The sequence of the cloned fragments in the vector constructs was determined, and the effect of constructs in MMCLs was examined using qRT-PCR and Western blot analysis.\nResults:\nIn this work, we established a method for making retroviral-shRNA expression constructs based on micro RNA-adapted short hairpin RNA (sh RNA mir) design. The HGF producing human myeloma cell line JJN-3 transduced with lentiviral HGF-shRNA transduction particles were analyzed for their effect on the HGF gene expression. HGF knockdown MMCL was analyzed both at mRNA level and protein levels. Two out of five clones of myeloma cell line JJN-3 (JJN3-sh3308 and JJN3-sh47137) were transduced with lentiviral HGF–shRNA transduction particles were showing 50-60 % knockdown in their HGF mRNA and protein expression.\nConclusion:\nHGF knockdown cell lines can be used as tissue culture models to investigate the role of HGF protein in the pathology of multiple myeloma. Further, the retroviral-HGF-shRNA expression constructs can study HGF response in various multiple myeloma via gene knockdown studies.</jats:p>","journal":"Journal of Khyber College of Dentistry","year":2020,"id":598304,"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":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1533007,"name":"Anders Sudan","orcid":null,"position":1,"is_corresponding":false},{"id":1533008,"name":"Asad Jan","orcid":null,"position":2,"is_corresponding":false},{"id":1533009,"name":"Shagufta Sultana","orcid":null,"position":3,"is_corresponding":false},{"id":1533006,"name":"Fouzia Qadir","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"ESTABLISHMENT OF MULTIPLE MYELOMA CELL  LINES WITH REDUCED EXPRESSION OF HEPATOCYTE  GROWTH FACTOR USING RNAI APPROACH","abstract":"<jats:p>Objective:\nTo establish the gene knockdown system (RNAi) using different Multiple Myeloma Cell lines (MMCL).\nMaterial and Methods:\nRNAi is generally used in biomedical research to study the outcome of knockdown of gene expression. In this research, retroviral-shRNA expression constructs based on micro RNA-adapted short hairpin RNA (sh RNA mir) were designed and constructed. The sequence of the cloned fragments in the vector constructs was determined, and the effect of constructs in MMCLs was examined using qRT-PCR and Western blot analysis.\nResults:\nIn this work, we established a method for making retroviral-shRNA expression constructs based on micro RNA-adapted short hairpin RNA (sh RNA mir) design. The HGF producing human myeloma cell line JJN-3 transduced with lentiviral HGF-shRNA transduction particles were analyzed for their effect on the HGF gene expression. HGF knockdown MMCL was analyzed both at mRNA level and protein levels. Two out of five clones of myeloma cell line JJN-3 (JJN3-sh3308 and JJN3-sh47137) were transduced with lentiviral HGF–shRNA transduction particles were showing 50-60 % knockdown in their HGF mRNA and protein expression.\nConclusion:\nHGF knockdown cell lines can be used as tissue culture models to investigate the role of HGF protein in the pathology of multiple myeloma. Further, the retroviral-HGF-shRNA expression constructs can study HGF response in various multiple myeloma via gene knockdown studies.</jats:p>","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"23304386","pmcid":null,"openalex_id":"https://openalex.org/W4385147205","authors":[],"funders":[],"total_grants":0,"fwci":0.0,"citation_percentile":0.21630943,"influential_citations":0,"citation_trend":[],"oa_status":"gold","license":"cc-by-nc-nd","oa_locations":[{"url":"https://journalofkcd.com/index.php/pmsrj/article/download/207/173","host_type":"journal"},{"url":"https://journalofkcd.com/index.php/pmsrj/article/download/207/173","host_type":"publisher"},{"url":"https://journalofkcd.com/kcd/article/download/207/173","host_type":"publisher"},{"url":"http://dx.doi.org/10.33279/jkcd.v10i03.207","host_type":"journal"}],"fields_of_study":["RNA Interference and Gene Delivery","Monoclonal and Polyclonal Antibodies Research","RNA and protein synthesis mechanisms"],"mesh_terms":[],"keywords":["Gene knockdown","Small hairpin RNA","RNA interference","Hepatocyte growth factor","Cell culture","Molecular biology","Biology","Gene expression","RNA","Gene","Cancer research","Cell biology","Genetics"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-28T15:27:54.185855Z","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":[]}