title: [Effect of blockade of NF-kappaB signaling pathway on cell apoptosis in cutaneous squamous cell carcinoma].
abstract: OBJECTIVE
  To evaluate the siRNA-mediated inhibitory effect of nuclear factor-kappaB (NF-kappaB) p65 on expression of p65, and explore the effect of blockade of NF-kappaB signal pathway on cell apoptosis in cutaneous squamous cell carcinoma (cutaneous SCC).
  
  
  METHODS
  Cutaneous SCC cell line SCL-1 cells were transfected with 50 nmol/L p65 siRNA. The expression level of p65 mRNA was measured using RT-PCR method at 0, 24, 48 and 72 h . Expressions of p65, bcl-2 and bax proteins were determined using Western blotting. Activities of caspase-3/9 was detected by Caspase-Glo®-3/7, 8 and 9 kit. Finally, cell apoptosis was detected using flow cytometry.
  
  
  RESULTS
  The expression level of p65 mRNA in Cutaneous SCC SCL-1 cells was obviously down-regulated 48 h after transfection with p65 siRNA, and a significant difference was detected, as compared with 0 h after (0.23 ± 0.10 vs. 0.66 ± 0.05, P<0.05). The protein levels of p65 and bcl-2 decreased, and the bax protein level and activities of caspase-3/9 increased after transfection with p65 siRNA at h 48 . Further, the results of flow cytometry demonstrated that p65 siRNA could induce apoptosis of SCL-1 cells, and cell apoptosis ratio (20.28% ± 1.87%) in p65 siRNA group was significantly higher than that in the untreated group and control siRNA group (9.13% ± 1.51% and 9.37% ± 1.38%, respectively, F=47.532, P<0.01).
  
  
  CONCLUSIONS
  p65 siRNA can block NF-kappaB signal pathway, down-regulate expression of bcl-2, elevate the bax level and increase the activities of caspase-3/9, suggesting that NF-kappaB signal pathway may be a key molecular target for therapy of cutaneous SCC.
authors.affiliation: Department of Dermatology, Xinxiang Medical University, Weihui 453100, China.
authors.forename: Yong-hua
authors.initials: YH
authors.lastname: Xia
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.forename: Cai-feng
authors.initials: CF
authors.lastname: Zhang
authors.forename: Dan-dan
authors.initials: DD
authors.lastname: Fu
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Zhan-guo
authors.initials: ZG
authors.lastname: Li
authors.forename: Zhong-wei
authors.initials: ZW
authors.lastname: Tian
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology*; D051996:Transcription Factor RelA / Q000096:biosynthesis / Q000235:genetics
chemical_list: D016328:NF-kappa B; D034741:RNA, Small Interfering; D051996:Transcription Factor RelA