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Effects of SPARC Gene Overexpression on Proliferation and Apoptosis of SKM-1 Cells
Nian Qing, Wei Chunmei, Huang Jing, Dan Chunli, Xiao Qing, Yang Zesong, Wang Li*
Department of Hematology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
Abstract: To construct a recombinant lentiviral vector carrying SPARC gene， we investigated the alteration on the proliferation of MDS cell line SKM-1 cells. The SPARC was obtained using pcDNA-SPARC by PCR. The SPARC gene was cloned into a lentiviral vector pGC-GV to construct a recombinant lentiviral vector carrying SPARC gene named pGC-GV-SPARC， which was confirmed to be correct by DNA sequencing. pGC-GV-SPARC was used to transfect human MDS cell line SKM-1. The transfection efficiency， SPARC mRNA and protein expression were detected by flow cytometry， RT-PCR and Western blot， respectively. The MTS method was used to determine inhibition effects of low doses of Ara-C in different groups. Results showed that the recombinant lentiviral vector pGC-GV-SPARC was confirmed to be correct by DNA sequencing. The transfection efficiency was (64.25±1.42)%， and stably expressed. PT-PCR and Western blot showed that the expression of SPARC was increased. MTS results showed that the proliferation effect of low doses of Ara-C on the inhibition rate of transfection group was significantly higher than those of negative group and SKM-1 group. The results showed that lentiviral vector carrying SPARC was constructed successfully， and the expression of SPARC could effectively increased by pGC-GV-SPARC， which could inhibit the proliferation of SKM-1 cell and promote its apoptosis by combining with low doses of Ara-C.
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