ESTABLISHMENT OF MULTIPLE MYELOMA CELL LINES WITH REDUCED EXPRESSION OF HEPATOCYTE GROWTH FACTOR USING RNAI APPROACH

Authors

  • Fouzia Qadir Department of Biochemistry North West School of Medicine Peshawar
  • Anders Sudan Department of Clinical and Molecular Medicine. Norwegian University of Science and Technology Trondheim Norway
  • Asad Jan Institute of Biotechnology and Genetic Engineering(IBGE), University of Agriculture Peshawar.
  • Shagufta Sultana Anatomy Department, Khyber Medical College, Peshawar.

DOI:

https://doi.org/10.33279/jkcd.v10i03.207

Keywords:

Multiple myeloma, Hepatocyte growth factor (HGF), Gene knockdown, short hairpin RNA

Abstract

Objective:

To establish the gene knockdown system (RNAi) using different Multiple Myeloma Cell lines (MMCL).

Material and Methods:

RNAi 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.

Results:

In 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.

Conclusion:

HGF 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.

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Published

2020-09-30

How to Cite

Fouzia Qadir, Anders Sudan, Asad Jan, & Shagufta Sultana. (2020). ESTABLISHMENT OF MULTIPLE MYELOMA CELL LINES WITH REDUCED EXPRESSION OF HEPATOCYTE GROWTH FACTOR USING RNAI APPROACH. JOURNAL OF KHYBER COLLEGE OF DENTISTRY, 10(03), 35–42. https://doi.org/10.33279/jkcd.v10i03.207

Issue

Vol. 10 No. 03 (2020): Journal of Khyber College of Dentistry

Section

Original Article

License

Copyright (c) 2020 Fouzia Qadir, Anders Sudan, Asad Jan, Shagufta Sultana

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