title: [FHL1 knockdown mediated by lentiviral shRNA promotes the growth of HeLa and HepG2 cells].
abstract: OBJECTIVE
  To investigate the effect of four-and-a-half LIM domain 1 (FHL1) knockdown by lentiviral-mediated shRNA on the growth of HeLa and HepG2 cells.
  
  
  METHODS
  pLenti-H1 FHL1 shRNA was cloned, and then transfered into HEK293T cells. The inhibitory effect of pLenti-H1 FHL1 shRNA on FHL1 gene was detected by Western blotting and real-time quantitative PCR (qRT-PCR). Lentivirus particles were packaged, added to HeLa and HepG2 cells, followed by puromycin treatment for 2-3 weeks to screen stable clones. The knockdown effect on FHL1 expression in these cells was checked by Western blotting and qRT-PCR. Cell growth and colony formation analysis were performed to investigate the effect of FHL1 down-regulation on tumor cell growth. Soft agar analysis was used to analyze its effect on anchorage-independent tumor cell growth.
  
  
  RESULTS
  Western blotting and qRT-PCR revealed that the pLenti-H1 FHL1 shRNA apparently inhibited the expression of FHL1 gene. Cell growth and colony formation assay showed that the lentiviral-mediated shRNA for FHL1 gene significantly accelerated the tumor cell growth in HepG2 and HeLa cells. Soft agar analysis demonstrated that FHL1 shRNA increased the anchorage-independent growth of tumor cells.
  
  
  CONCLUSIONS
  pLenti-H1 FHL1 shRNA could significantly accelerate tumor cell growth via inhibiting the expression of FHL1.
authors.affiliation: Department of Medical Oncology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medicine, Beijing 100850, China.
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Gastroenterology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medicine, Beijing 100850, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Endocrine, Beijing Military 264 Hospital, Taiyuan 030001, China.
authors.forename: Baiyu
authors.initials: B
authors.lastname: Han
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medicine, Beijing 100850, China.
authors.forename: Yingchun
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medicine, Beijing 100850, China.
authors.forename: Qinong
authors.initials: Q
authors.lastname: Ye
authors.affiliation: Department of Medical Oncology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Shunchang
authors.initials: S
authors.lastname: Jiao
mesh_terms: D049109:Cell Proliferation / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000172:deficiency* / Q000235:genetics*; D060588:LIM Domain Proteins / Q000172:deficiency* / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics*; D009124:Muscle Proteins / Q000172:deficiency* / Q000235:genetics*; D014411:Neoplastic Stem Cells / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: C101149:FHL1 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D009124:Muscle Proteins; D034741:RNA, Small Interfering