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Overexpression of MFRN1 in 293T Cell and Its Effects on the Cell Proliferation and Mitochondrial Function
Yang Yu1, Chen Lin1, Zhang Xiaoqin1, Zhang Liu1, Zhang Wanlin1, Shi Mengru1,2,3, LV Jianxin1,2,3*, Li Wei1,2,3*
1Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou 325035, China; 2Key Laboratory of Laboratory Medicine, Ministry of Education, Wenzhou 325035, China; 3School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035, China
Abstract: Mitoferrin-1 (MFRN1) is a member of the solute carrier family as an essential iron importer， which is located in the mitochondrial inner membrane. In order to investigate the effect of MFRN1 overexpression in 293T cell line on cell proliferation and mitochondrial function， a MFRN1 stably overexpressed cell line was generated by lentiviral-mediated MFRN1 transfected into 293T cell line， then the mitochondrial function was measured. It was found that the level of MFRN1 expression was significantly increased in the MFRN1 stably overexpressed 293T cell line (P<0.01). Furthermore， it was observed that the total iron loading was markedly elevated in the overexpressed cell line (P<0.05). In addition， it was also found that the mitochondrial structure was damaged with mitochondrial cristae reduced and vacuolated， as well as energy metabolism in mitochondrion was inhibited. It was indicated that the cellular ATP content markedly decreased and total adenine nucleotide pool reduced by 40% compared to controls. Even tually， it was showed that the proliferation of overexpressed cell line were inhibited with significant reduction of cell counts， whereas the content of cellular hydroxyl free radical (·OH) increased and oxidative damage of mitochondrial DNA occurred. It was concluded that MFRN1 played important roles in the cellular function， and the aberrant expression of MFRN1 could result in the mitochondrial dysfunction and inhibition of cell proliferation.
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