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Hepatocellular Carcinoma Specific Transcriptional Interference of c‐Myc promoter by alpha‐fetoprotein and Sendai Virosome Based dsRNA System
Author(s) -
Zakaria Mohammad Khalid,
Khan Imran,
Mani Prashant,
Chattopadhyay Parthaprasad,
Sarkar Debi P.,
Sinha Subrata
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.lb115
Subject(s) - small hairpin rna , hepatocellular carcinoma , biology , gene silencing , cancer research , rna interference , enhancer , carcinogenesis , microbiology and biotechnology , sendai virus , promoter , epigenetics , gene , virology , rna , transcription factor , gene expression , virus , genetics
Objective c‐Myc is one of the central players of oncogenesis in hepatocellular carcinoma (HCC) and is also required in normal development. Majority of the c‐Myc transcripts are transcribed by its P2 promoter and an upstream ME1a1 site is required in its transcriptional activation. dsRNA also recruits epigenetic silencing complexes inducing long term/heritable transcriptional repression. We devised a novel HCC targeting system and studied its mechanism. Methods Firstly, the alpha‐fetoprotein (AFP) promoter/enhancer system was used to express shRNA against ME1a1 site. Secondly, we transformed Sendai virus into fusion (F) virosomes for packaging and delivery of AFP promoter/enhancer driven shRNA constructs into liver cells. Results Following F‐virosomal delivery, shRNA induced heterochromatization (H3K9Me2 and H3K27Me3) and CpG 8,9,10 methylation at the target locus. Following c‐Myc suppression, HCC cells showed decreased cell survival and increased apoptosis. Also, c‐Myc suppressed HCC cells were sensitized to sub lethal doses of 5‐Fluorouracil (5‐FU) and Cisplatin. Conclusion This system can specifically and prominently repress c‐Myc in HCC cells and could also be used to introduce genes specifically in embryonic liver. Grant Funding Source : Department of Biotechnology, Govt. of India