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Tumor suppressor genes are differentially regulated with dietary folate modulations in a rat model of hepatocellular carcinoma

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Abstract

The current study evaluated the outcome of dietary folate modulations on the expression of tumor suppressor genes (TSGs) during developmental stages of hepatocellular carcinoma (HCC) in a Wistar rat model. In addition to dietary folate modulations, male rats were administered diethylnitrosamine (DEN) intraperitoneally once a week upto 18 weeks to induce HCC. Serum folate levels were found to be decreased and increased in folate deficiency (FD) and folate-oversupplemented (FO) groups respectively when compared to folate normal (FN) rats. Apoptosis was observed in FD in fibrosis and HCC stages. mRNA expression analysis by RT-PCR of TSGs (DPT, p16, RUNX3, RASSF1A and SOCS1) and protein expression by western blot (RASSF1A, RUNX3 and p16) depicted differential expression in FD and FO in various stages of HCC development. Bisulfite sequencing for p16 and RASSF1A promoter was performed. The promoter region of p16 gene was hypermethylated at 7th and that of RASSF1A was hypomethylated at 10th CpG in cirrhotic category in FD rats. Hyper and hypomethylation at 10th and 24th CpG respectively in RASSF1A promoter was observed in HCC category in both FD and FO groups. All TSGs showed differential expression at transcript and protein level. Increased expression of DPT, RASSF1A, SOCS1 and decreased expression of RUNX3 could be playing role in HCC development in FD rats. Reduced expression of RUNX3, RASSF1A and SOCS1 in HCC category was demonstrated in FO rats. Thus, the studied TSGs are differentially expressed with dietary folate modulations during the development of HCC in DEN-treated rat model and the promoter methylation might be a contributing mechanism under these conditions.

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Abbreviations

FN:

Folate normal

FD:

Folate deficiency

FO:

Folate over-supplementation

TSG:

Tumor suppressor genes

DPT:

Dermatopontin

RUNX3:

Runt-related transcription factor

RASSF1A:

Ras association domain family member 1

SOCS1:

Suppressor of cytokine signaling1

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Authors and Affiliations

  1. Department of Biochemistry, Post Graduate Institute of Medical Education and Research, Chandigarh, India

    Renuka Sharma, Taqveema Ali & Jyotdeep Kaur

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  1. Renuka Sharma
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  2. Taqveema Ali
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  3. Jyotdeep Kaur
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Contributions

JK and RS has designed the overall study. RS has conducted hands on experiments. TA has contributed in the acquisition, and analysis of data related to blotting experiments. Indian Council of Medical Research provided funding to the research scholar (RS).

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Correspondence to Jyotdeep Kaur.

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Sharma, R., Ali, T. & Kaur, J. Tumor suppressor genes are differentially regulated with dietary folate modulations in a rat model of hepatocellular carcinoma. Mol Cell Biochem 476, 385–399 (2021). https://doi.org/10.1007/s11010-020-03915-3

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