Patrick C Flannery1,2, Kodye L Abbott1,2, Satyanarayana R Pondugula3,4. 1. Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University, 109 Greene Hall, Auburn, AL, 36849, USA. 2. Auburn University Research Initiative in Cancer, Auburn University, Auburn, AL, 36849, USA. 3. Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University, 109 Greene Hall, Auburn, AL, 36849, USA. srp0010@auburn.edu. 4. Auburn University Research Initiative in Cancer, Auburn University, Auburn, AL, 36849, USA. srp0010@auburn.edu.
Abstract
BACKGROUND AND OBJECTIVE: In many patients with hepatocellular carcinoma (HCC), cytochrome P450 3A4 (CYP3A4) expression has been reported to be significantly reduced in the tumor liver tissue. Moreover, this CYP3A4 repression is associated with decreased CYP3A4-mediated drug metabolism in the tumor liver tissue. However, the underlying mechanisms of CYP3A4 repression are not fully understood. We have previously shown that Mg2+/Mn2+-dependent phosphatase 1A (PPM1A) positively regulates human pregnane X receptor (hPXR)-mediated CYP3A4 expression in a PPM1A expression-dependent manner. We sought to determine whether PPM1A expression is downregulated and whether PPM1A downregulation is correlated with CYP3A4 repression in the tumor liver tissue of HCC patients. METHODS: Quantitative RT-PCR and western blot analyses were performed to study mRNA and protein expression, respectively. Cell-based reporter gene assays were conducted to examine the hPXR transactivation of CYP3A4 promoter activity. RESULTS: Arginase-1 and glypican-3 gene expression studies confirmed that the tumor samples used in our study originate from HCC livers but not non-hepatocellular tumors. mRNA and protein expression of PPM1A and CYP3A4 was found to be significantly repressed in the tumor liver tissues compared to the matched non-tumor liver tissues. In the reporter gene assays, elevated PPM1A levels counteracted the inhibition of hPXR-mediated CYP3A4 promoter activity by signaling pathways that are upregulated in HCC, suggesting that decreased PPM1A levels in HCC could not fully counteract the hPXR-inhibiting signaling pathways. CONCLUSIONS: Together, these results are consistent with the conclusion that PPM1A downregulation in the tumor liver tissue of HCC patients correlates with CYP3A4 repression. Downregulation of PPM1A levels in the tumor liver tissue may contribute to reduced hPXR-mediated CYP3A4 expression, and provide a novel mechanism of CYP3A4 repression in the tumor liver tissue of HCC patients.
BACKGROUND AND OBJECTIVE: In many patients with hepatocellular carcinoma (HCC), cytochrome P450 3A4 (CYP3A4) expression has been reported to be significantly reduced in the tumor liver tissue. Moreover, this CYP3A4 repression is associated with decreased CYP3A4-mediated drug metabolism in the tumor liver tissue. However, the underlying mechanisms of CYP3A4 repression are not fully understood. We have previously shown that Mg2+/Mn2+-dependent phosphatase 1A (PPM1A) positively regulates humanpregnane X receptor (hPXR)-mediated CYP3A4 expression in a PPM1A expression-dependent manner. We sought to determine whether PPM1A expression is downregulated and whether PPM1A downregulation is correlated with CYP3A4 repression in the tumor liver tissue of HCCpatients. METHODS: Quantitative RT-PCR and western blot analyses were performed to study mRNA and protein expression, respectively. Cell-based reporter gene assays were conducted to examine the hPXR transactivation of CYP3A4 promoter activity. RESULTS:Arginase-1 and glypican-3 gene expression studies confirmed that the tumor samples used in our study originate from HCC livers but not non-hepatocellular tumors. mRNA and protein expression of PPM1A and CYP3A4 was found to be significantly repressed in the tumor liver tissues compared to the matched non-tumor liver tissues. In the reporter gene assays, elevated PPM1A levels counteracted the inhibition of hPXR-mediated CYP3A4 promoter activity by signaling pathways that are upregulated in HCC, suggesting that decreased PPM1A levels in HCC could not fully counteract the hPXR-inhibiting signaling pathways. CONCLUSIONS: Together, these results are consistent with the conclusion that PPM1A downregulation in the tumor liver tissue of HCCpatients correlates with CYP3A4 repression. Downregulation of PPM1A levels in the tumor liver tissue may contribute to reduced hPXR-mediated CYP3A4 expression, and provide a novel mechanism of CYP3A4 repression in the tumor liver tissue of HCCpatients.
Authors: Kodye L Abbott; Julia M Salamat; Patrick C Flannery; Chloe S Chaudhury; Aneesh Chandran; Saraswathi Vishveshwara; Sridhar Mani; Jianfeng Huang; Amit K Tiwari; Satyanarayana R Pondugula Journal: ACS Omega Date: 2022-09-13