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Expression and Antitumor Function of Latcripin-4 RCC1 and ANK Domain Protein on HepG2 from the Shiitake Medicinal
Xuefang Guo1, Xiaoli Wang1, Min Liu1
1Department of Medical Microbiology, Dalian Medical University, Lvshunkou District, Dalian, China.
Abstract:
The recombinant protein of Latcripin-4 regulator of chromosome condensation 1 (RCC1) and ankyrin (ANK) domains were expressed and the antitumor activity of Latcripin-4 on HepG2 cells was studied. First, the Latcripin-4 transcript was selected from the medicinal mushroom Lentinus edodes C91-3 transcriptome by bioinformatics. Then the full-length gene of Latcripin-4 was isolated with 3'-full rapid amplification of cDNA ends (RACE) and 5'-full RACE methods according to the transcriptome. The RCC1 and ANK domains from the full-length gene were selected and inserted into the expression vector pET-32a (+) and expressed in Escherichia coli Rosetta-gami (DE3). Western blotting indicated that the protein was expressed successfully. The biological function of Latcripin-4 RCC1 and ANK domain protein on HepG2 cells was studied with the CCK-8 assay. All results demonstrated that Latcripin-4 RCC1 and ANK domain protein can inhibit the growth of human HepG2 liver cancer cells, which brings new insights to identifying antitumor proteins from medicinal food for cancer therapy.
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