• info@icbcongress.com
• English
• Persian

• Login

Create Free Account

INTERNATIONAL CONGRESS OF

CancerGenetics

• 1. Investigation of lncRNA HULC Expression in Reovirus-Infected Huh-7 Cells

• Reihaneh Kazemi,^1,* Mojtaba Hamidifard,² Elham Siasi,³ Mohammad Reza Aghasadeghi,⁴ Pooneh Rahimi,⁵
  1. Department of Genetics, NT.C., Islamic Azad University, Tehran, Iran
  2. Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran
  3. Department of Genetics, NT.C., Islamic Azad University, Tehran, Iran
  4. Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran
  5. Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran
  
  
  
• Introduction: Reoviruses are oncolytic viruses that can destroy different cancer cells, such as HCC. Understanding the dynamics of reovirus replication and the host cellular response is crucial for developing antiviral strategies and exploring the potential oncolytic properties of reoviruses. Long non-coding RNAs (lncRNAs) have emerged as important regulators of gene expression and cellular stress responses during viral infections. Among these, the highly upregulated in liver cancer (HULC) lncRNA has been implicated in modulating cellular pathways related to proliferation, apoptosis, and immune responses. However, the interaction between reovirus infection and HULC expression remains poorly understood. In this study, we investigated the temporal pattern of reovirus replication in Huh-7 cells and examined the corresponding changes in HULC expression. Our findings shed light on the complex interplay between viral replication and host lncRNA regulation, providing new insights into cellular defense mechanisms and potential therapeutic targets.
• Methods: Huh-7 cells were cultured in 6-well plates containing DMEM supplemented with 10% FBS. Once a monolayer formed, the culture medium was removed, and the cells were washed with PBS. The cells were then infected with reovirus at a multiplicity of infection (MOI) of 5. Cell samples were collected at various time points post-infection (0, 2, 4, 6, 24, 48, and 72 hours). Uninfected cells were used as controls. Total RNA extraction and cDNA synthesis were performed following the manufacturer’s instructions. Real-time PCR was conducted using 2x SYBR Green master mix to evaluate reovirus RNA replication and lncRNA HULC expression. Each run included a negative control and was performed in duplicate. The human β-globin gene served as an internal control. The specificity of the PCR products was confirmed by melting curve analysis.
• Results: Real-time PCR results showed that the rate of reovirus replication increased with longer infection times. The expression level of HULC was elevated in the 24- and 48-hour samples compared to the control; however, at 72 hours, its expression decreased below that of the control sample.
• Conclusion: Our study demonstrates that reovirus infection in Huh-7 cells leads to a time-dependent increase in viral RNA replication, accompanied by dynamic changes in the expression of the lncRNA HULC. The observed increase in HULC expression at 24 and 48 hours post-infection likely reflects a cellular response to the initial viral stress induced by mild reovirus replication. During this phase, cells activate defense mechanisms to enhance their survival, including upregulation of HULC expression. However, by 72 hours post-infection, extensive viral replication results in significant cell death and lysis. This advanced stage of infection leads to the degradation of various RNA molecules, including mRNAs and long non-coding RNAs such as HULC. These findings provide valuable insights into the interplay between reovirus replication and host lncRNA regulation, which may inform future therapeutic strategies targeting viral infections and host responses.
• Keywords: Reovirus, lncRNA Hulk, Real-time PCR.