Posters
Presenting Author Academic/Professional Position
Yamile Abuchard Anaya
Academic Level (Author 1)
Medical Student
Academic Level (Author 3)
Post-doc
Discipline/Specialty (Author 3)
Cancer and Immunology
Academic Level (Author 4)
Faculty
Discipline/Specialty (Author 4)
Cancer and Immunology
Discipline Track
Biomedical Science
Abstract Type
Research/Clinical
Abstract
Background: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and disproportionately affects underserved populations due to socioeconomic and healthcare disparities. The Rio Grande Valley (RGV) in South Texas exemplifies this issue, with high rates of obesity, diabetes, non-alcoholic fatty liver disease (NAFLD), and viral hepatitis (B and C)—all major risk factors for HCC. McAllen, Texas, part of the RGV, is notably one of the most obese cities in the U.S., exacerbating HCC prevalence and severity in this region. Late-stage diagnosis remains a critical challenge due to limited access to screening and early intervention.
Sorafenib, a first-line tyrosine kinase inhibitor (TKI), remains a cornerstone of therapy for advanced HCC, yet resistance frequently develops. Y-box binding protein 1 (YBX1), a 324-amino acid member of the cold shock domain protein family, plays a key role in mRNA translation and interacts with non-coding RNAs and microRNAs. Its involvement in drug resistance and aggressive cancer behavior makes it a compelling molecular target for further investigation.
Methods: The HCC cell line SK-Hep1 was cultured under standard ATCC conditions. Lentiviral vectors encoding YBX1 (GFP-tagged with puromycin resistance) were used to create stable overexpression and knockdown models. Both monoclonal (single-cell-derived) and polyclonal (population-level) cell lines were established to evaluate clonal effects on YBX1 function. YBX1 expression was confirmed through Western blotting and RT-PCR. To analyze functional outcomes, a series of assays were performed, including migration, invasion, proliferation, colony formation, and impedance-based growth analysis using the xCELLigence system.
Results: TCGA analysis confirmed that YBX1 is significantly upregulated in HCC tumors compared to normal tissue and is linked to poor prognosis, metastasis, and disease progression. In vitro, YBX1 overexpression enhanced oncogenic properties, including migration, invasion, proliferation, and colony formation, while YBX1 knockdown suppressed these features. Impedance analysis showed increased real-time proliferation in YBX1-overexpressing cells and a decreased growth rate in knockdown lines. Monoclonals were systematically developed from the overexpression and knockdown of cell lines. Notably, monoclonal YBX1 OE and KD exhibited phenotypes consistent with their expression levels. While polyclonal populations showed variability in response to YBX1 modulation, suggesting that a clonally selected population might have a more consistent dose-dependent response. This variation could influence YBX1 signaling and drug resistance profiles.
Conclusion and future Directions: We successfully developed and validated SK-Hep1 cell clones with systematically engineered overexpression (OE) and knockdown (KD) of YBX1. The selected clones provide valuable tools for studying oncogenic signaling, therapeutic resistance, and the crucial role of YBX1 in promoting the aggressiveness of HCC. Insights from these models could inform the development of precision-targeted therapies for HCC, particularly in underserved regions where effective treatment options are scarce.
Presentation Type
Poster
Recommended Citation
Abuchard Anaya, Yamile; Renteria, Kaylee; Nagati, Veerababu; and Tripathi, Manish K., "Comparative Oncogenic Phenotypes of Polyclonal and Monoclonal YBX1-Expressing HCC Cell Lines" (2025). Research Colloquium. 111.
https://scholarworks.utrgv.edu/colloquium/2025/posters/111
Included in
Comparative Oncogenic Phenotypes of Polyclonal and Monoclonal YBX1-Expressing HCC Cell Lines
Background: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and disproportionately affects underserved populations due to socioeconomic and healthcare disparities. The Rio Grande Valley (RGV) in South Texas exemplifies this issue, with high rates of obesity, diabetes, non-alcoholic fatty liver disease (NAFLD), and viral hepatitis (B and C)—all major risk factors for HCC. McAllen, Texas, part of the RGV, is notably one of the most obese cities in the U.S., exacerbating HCC prevalence and severity in this region. Late-stage diagnosis remains a critical challenge due to limited access to screening and early intervention.
Sorafenib, a first-line tyrosine kinase inhibitor (TKI), remains a cornerstone of therapy for advanced HCC, yet resistance frequently develops. Y-box binding protein 1 (YBX1), a 324-amino acid member of the cold shock domain protein family, plays a key role in mRNA translation and interacts with non-coding RNAs and microRNAs. Its involvement in drug resistance and aggressive cancer behavior makes it a compelling molecular target for further investigation.
Methods: The HCC cell line SK-Hep1 was cultured under standard ATCC conditions. Lentiviral vectors encoding YBX1 (GFP-tagged with puromycin resistance) were used to create stable overexpression and knockdown models. Both monoclonal (single-cell-derived) and polyclonal (population-level) cell lines were established to evaluate clonal effects on YBX1 function. YBX1 expression was confirmed through Western blotting and RT-PCR. To analyze functional outcomes, a series of assays were performed, including migration, invasion, proliferation, colony formation, and impedance-based growth analysis using the xCELLigence system.
Results: TCGA analysis confirmed that YBX1 is significantly upregulated in HCC tumors compared to normal tissue and is linked to poor prognosis, metastasis, and disease progression. In vitro, YBX1 overexpression enhanced oncogenic properties, including migration, invasion, proliferation, and colony formation, while YBX1 knockdown suppressed these features. Impedance analysis showed increased real-time proliferation in YBX1-overexpressing cells and a decreased growth rate in knockdown lines. Monoclonals were systematically developed from the overexpression and knockdown of cell lines. Notably, monoclonal YBX1 OE and KD exhibited phenotypes consistent with their expression levels. While polyclonal populations showed variability in response to YBX1 modulation, suggesting that a clonally selected population might have a more consistent dose-dependent response. This variation could influence YBX1 signaling and drug resistance profiles.
Conclusion and future Directions: We successfully developed and validated SK-Hep1 cell clones with systematically engineered overexpression (OE) and knockdown (KD) of YBX1. The selected clones provide valuable tools for studying oncogenic signaling, therapeutic resistance, and the crucial role of YBX1 in promoting the aggressiveness of HCC. Insights from these models could inform the development of precision-targeted therapies for HCC, particularly in underserved regions where effective treatment options are scarce.
