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A Preliminary Study on the Regulation of Hepatoma Cell Biological Behavior by MCFP

Author: ZuoZuo
Tutor: WangSiYing;YangXiaoMing
School: Anhui Medical University,
Course: Pathology and Pathophysiology
Keywords: lentiviral vector MCFP RNA interference Human hepatocellular liver
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 4
Quote: 0
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Abstract


In eukaryotic cell mitochondria is the main site of energy metabolism. Mitochondrial function depends on a variety of soluble substances across the mitochondrial inner membrane transport of the endometrium, which is mainly completed by the transfer of solute carrier (SLC). SLC located in the inner mitochondrial membrane to form mitochondrial transporter family, including SLC25 family. The SLC25 family members had found a total of 14 species, of which SLC25A40, also known as MCFP (mitochondrial carrier functional protein), is a new member which identified in the central nervous system. MCFP as a newly discovered gene, the research reported rarely home and abroad.In our research we found that the expression of MCFP in most liver cancer cell lines was highter than normail liver cells. In our pervious research the effective sequence of siRNA targeting MCFP gene was confirmed, the complementary DNA containing both sense and antisense Oligo DNA of targeting sequence was designed, synthesized and cloned into the pSiCoR vector. The resulting lentiviral vector containing MCFP shRNA was named pSiCoR–MCFP. The MCFP cell lines were transfected by pSiCoR -MCFP recombinant lentivirus and the stable transfected liver cancer cell lines and normal liver cell line were established. The interference effect of MCFP was analyzed by PCR and Western blot. They both confirmed that the expression of MCFP decreased in the stable transfected HepG2 cell line (HepG2-SiMCFP VS HepG2-Control, P <0.001). Using the stable HepG2 cell lines we detected changes of the cell behavior and the mitochondrial function. Conclusion that knockdown MCFP in HepG2 cells can suppress cell proliferation, hold back cell course at S, and accelerate cell apoptosis. Also the production of ATP reduced and the complex activity decreased significantly. In our research we detected changes of the cell behavior and the mitochondrial function in the stable transfect HepG2 cell line. It can be used for further study of the role of the MCFP gene in the regulation of HepG2 cell. In conclusion, we investigated biological function of MCFP, opening up new avenues of inquiry into mitochondrial protein functions and mitochondrial disease mechanism.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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