title: [Down-regulation of ubiquitin using short hairpin RNA inhibits the proliferation and promotes apoptosis of lung cancer H1299 cells].
abstract: OBJECTIVE
  To study the effect of the down-regulation of ubiquitin gene on the proliferation of human lung cancer cells, and explore its probable mechanism.
  
  
  METHODS
  Plasmids shRNA-UBB and shRNA-UBC were designed and used to knock down the expression of ubiquitin gene (Ub-KD) in human lung cancer H1299 cells. The silence effect was verified by RT-PCR and Western blotting. The effect of Ub-KD on the proliferation of H1299 cells was evaluated using the MTT assay and plate clone formation assay. Cell apoptosis was detected by annexin V-PE staining and DNA fragment assay. Cell cycle distribution was assessed by flow cytometry and PI staining.
  
  
  RESULTS
  The MTT assay showed that Ub-KD (shRNA-UBB/UBC) significantly decreased the proliferation of H1299 cells after 48- and 72-hour silencing. The clone formation assay showed that Ub-KD (shRNA-UBB/UBC) decreased the colony forming ability when compared with shRNA-NC group. Compared with shRNA-NC group, Ub-KD (shRNA-UBB/UBC) promoted the cell apoptosis (shRNA-NC, 5.60%; shRNA-UBB, 9.07%; shRNA-UBC, 11.70%; shRNA-UBB/UBC, 17.82%). Compared with shRNA-NC group, Ub-KD (shRNA-UBB/UBC) increased the G1 phase arrest significantly (shRNA-NC, 34.53%; shRNA-UBB, 34.40%; shRNA-UBC, 42.77%; shRNA-UBB/UBC, 50.20%).
  
  
  CONCLUSIONS
  Down-regulation of ubiquitin gene via Ub-KD (shRNA-UBB combined with shRNA-UBC) may inhibit the proliferation of lung cancer cells by promoting the cell apoptosis and increasing the G1 phase arrest.
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Yiting
authors.initials: Y
authors.lastname: Tang
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Wenhao
authors.initials: W
authors.lastname: Shen
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: He
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Shuyu
authors.initials: S
authors.lastname: Zhang
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Jinchang
authors.initials: J
authors.lastname: Wu
authors.affiliation: School of Radiation Medicine and Protection, Medical College, Soochow University, Jiangsu Key Laboratory of Radiation Medicine and Protection, Suzhou 215123, China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Cao
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation; D059585:G1 Phase Cell Cycle Checkpoints; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology / Q000628:therapy*; D034741:RNA, Small Interfering / Q000235:genetics*; D025801:Ubiquitin / Q000037:antagonists & inhibitors* / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering; D025801:Ubiquitin