5333
Heba Kamal Nabih Hanafi
The Emerging Role of MicroRNA 122 in Intracellular
Signaling Interactions of Hepatocellular Carcinoma
Hepatocellular carcinoma- Multidrug resistance- MicroRNA
122- Doxorubicin- MAPK pathway- QRT-PCR- Western blotting- ELISAFlow
cytometry- Alpha-fetoprotein.
Background: Hepatocellular carcinoma (HCC) is the leading primary liver cancer and is considered the third frequent cause of cancer-related death worldwide. Curative treatments, including liver resection, transplantation, and local ablation, could provide promising 5-year-survival rate up to 75%, nevertheless, lower than twenty percent of HCC patients are qualified for these treatments. Furthermore, systemic chemotherapeutic options for HCC are limited and failure because of multidrug resistance (MDR) development. Thus, there is a need for new targeted treatments that have a role in the regulation of tumor cell sensitivity to drugs and modulating the expression of genes/proteins implicated in MDR.Material and Methods: Human HCC-cell line (HepG2 cells) was cultured and subjected to different treatments of doxorubicin and/or microRNA-122 (mir-122) mimics. Total microRNAs were isolated for estimation of mir-122 overexpression. For quantification of MDR-related genes/proteins, total RNAs and protein extracts were isolated for application of QRT- PCR and western blotting techniques, respectively. As well as, genes of stressactivated MAPK pathways were detected by QRT-PCR. Also, cell cycle distribution was analyzed by flowcytometry. In addition, cell culture media were isolated for detection of alpha- fetoprotein by ELISA.
2017
Ph.d
Ain Shams
Science