Effects of mitofusin-2 gene on cell proliferation and chemotherapy sensitivity of MCF-7.
- Author:
Yun, XIA
;
Yaqun, WU
;
Xiaojun, HE
;
Jianping, GONG
;
Fazu, QIU
- Publication Type:Journal Article
- MeSH:
Antineoplastic Agents, Phytogenic/pharmacology;
Apoptosis;
Camptothecin/pharmacology;
Cell Cycle;
Cell Line, Tumor;
Cell Proliferation;
Drug Screening Assays, Antitumor;
Flow Cytometry;
Gene Expression Regulation, Neoplastic;
Green Fluorescent Proteins/metabolism;
Membrane Proteins/*biosynthesis;
Membrane Proteins/*genetics;
Mitochondrial Proteins/*biosynthesis;
Mitochondrial Proteins/*genetics;
Transfection
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2008;28(2):185-9
- CountryChina
- Language:English
-
Abstract:
In order to evaluate the effect of mitofusin-2 gene (mfn2) on proliferation and chemotherapy sensitivity of human breast carcinoma cell line MCF-7 in vitro, pEGFPmfn2 plasmid carrying full length of mitofusin-2 gene was transfected, by using sofast, into MCF-7 cells. Mitofusin-2 gene expression in MCF-7 cells transfected by sofast after 48 h was detected by PCR and Western blotting, and the stable expression of GFP protein in MCF-7 cells by Western blot analysis. The proliferation of MCF-7 cells was assayed by MTT and cell counting. By using PI method, the effects of mfn2 on the cell cycle distribution of MCF-7 were measured. Annexin-V/PI double labeling method was employed to detect the changes in apoptosis induced by chemotherapeutics before and after transfection. The results showed that the MCF-7 cells transfected with mfn2 gene could stably and highly express GFP protein. MTT assay revealed that after transfection of mfn2 cDNA, the proliferation of MCF-7 cells was significantly inhibited. DNA histogram showed that cells arrested in S phase, and the percentage of S phase cells was 42.7, 17.2 and 19.6 in mfn2 cDNA transfection group, blank plasmid transfection group and blank control group, respectively (P<0.05). The apoptosis ratio of the cells transfected with mfn2 gene was increased from 3.56% to 15.95%, that of the cells treated with camptothecin (CAMP) followed by mfn2 gene transfection was 69.6%, and that in blank plasmid transfection group and blank control group was 31.0% and 23.4% respectively (P<0.05). It was suggested that transfection of mfn2 gene could significantly inhibit the proliferation of MCF-7 cells and promote their sensitivity to CAMP with a synergic effect.
