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Author:

Wang, S. L. (Wang, S. L..) | Deng, W. T. (Deng, W. T..) | Wen, G. F. (Wen, G. F..) | Li, C. W. (Li, C. W..) | Zeng, Y. J. (Zeng, Y. J..)

Indexed by:

Scopus SCIE

Abstract:

OBJECTIVE: The aim of this study was to construct a eukaryotic expression plasmid with a short hairpin RNA (shRNA) targeting Livin in order to obtain a stably transfected Hep-2 cell line with a reduced expression of Livin. METHODS: The shRNA targeting Livin mRNA was designed, and a shRNA plasmid and a negative control plasmid were constructed. After amplification in E. coli, restriction endonuclease digestion and sequence confirmation, the plasmids were transfected into Hep-2 cells using Lipofectamine 2000. The stably transfected cell line was screened using G418, and inhibition of Livin mRNA and protein levels were detected using real-time PCR and western blotting, respectively. RESULTS: pGenesil-Livin-shRNA eukaryotic expression plasmid was successfully constructed and identified by sequencing. Green fluorescent protein (GFP) expression was observed in Hep-2 cells transfected with shRNA plasmids by fluorescence microscopy. The expression levels of Livin mRNA and protein decreased significantly in Hep-2 cells transfected with the shRNA recombinant plasmid. The mRNA level was reduced by 47.17 %, and the protein level was reduced by 34.25 %. CONCLUSION: The shRNAeukaryotic expression plasmid targeting Livin was successfully constructed, which could significantly inhibit the expression of Livin in laryngeal cancer Hep-2 cells. This provides a basis for future research on the function of Livin in Hep-2 cells, and gene therapy for laryngeal cancer (Fig. 5, Ref. 10). Text in PDF www.elis.sk.

Keyword:

RNA interference apoptosis regulatory proteins cell culture livin gene

Author Community:

  • [ 1 ] [Wang, S. L.]Shantou City Cent Hosp, Dept Otorhinolaryngol Head & Neck Surg, Shantou, Peoples R China
  • [ 2 ] [Deng, W. T.]Shantou City Cent Hosp, Dept Otorhinolaryngol Head & Neck Surg, Shantou, Peoples R China
  • [ 3 ] [Wen, G. F.]Shantou City Cent Hosp, Dept Otorhinolaryngol Head & Neck Surg, Shantou, Peoples R China
  • [ 4 ] [Li, C. W.]Shantou City Cent Hosp, Dept Otorhinolaryngol Head & Neck Surg, Shantou, Peoples R China
  • [ 5 ] [Zeng, Y. J.]Beijing Univ Technol, Biomech & Med Informat Inst, Beijing, Peoples R China

Reprint Author's Address:

  • [Wang, S. L.]Shantou City Cent Hosp, Dept Otorhinolaryngol Head & Neck Surg, Shantou, Peoples R China

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Source :

BRATISLAVA MEDICAL JOURNAL-BRATISLAVSKE LEKARSKE LISTY

ISSN: 0006-9248

Year: 2016

Issue: 5

Volume: 117

Page: 272-275

1 . 5 0 0

JCR@2022

ESI Discipline: CLINICAL MEDICINE;

ESI HC Threshold:203

CAS Journal Grade:4

Cited Count:

WoS CC Cited Count: 2

SCOPUS Cited Count: 2

ESI Highly Cited Papers on the List: 0 Unfold All

WanFang Cited Count:

Chinese Cited Count:

30 Days PV: 10

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