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Mitofusin-2 gene enhances sensitivity of human breast cancer T47D cells to parthenolide
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Abstract:
Objective:To investigate the effect of mitofusin-2 ( Mfn-2 ) gene expression on sensitivity of human breast cancer T47D cells to parthenolide. Methods: The expressions of Mfn-2 mRNA in various breast cancer cell lines (T47D, MDA-MB-231, MCF-7, MDA-MB-435 and HCC38) were detected by real-time PCR. Plasmids pEGFP and pEGFP-Mfn-2 were transfected into human breast cancer T47D cells by LipofectamineTM 2000 in vitro. The expression levels of Mfn-2 mRNA and protein in T47D cells were detected by real-time PCR and Western blotting. MTT assay was used to detect the proliferation of T47D cells. The cell apoptotic rate and mitochondrial membrane potential of T47D cells were measured by flow cytometry. Results: Compared with that in normal breast cells, Mfn-2 mRNA was highly expressed in breast cancer HCC38 cell line, and lowly expressed in the other cell lines, such as T47D etc. After pEGFP-Mfn-2 transfection for 48 h, the expression levels of Mfn-2 mRNA and rotein were significantly up-regulated in T47D cells. Compared with the pEGFP transfection group, the pEGFP-Mfn-2 transfection group showed a significant decrease in surivival rate of T47D cells under the treatment of parthenolide (50 mmol/L) (\[47.93±2.21\]% vs \[56.93±2.05\]%, P<0.05). Flow cytometry results showed that the apoptotic rate of T47D cells under the treatment of 0.05 mol/L parthenolide was significantly increased in pEGFP-Mfn-2 transfection group compared with that in pEGFP ransfection group (\[71.2±2.1\] % vs \[38.8±2.6\] %, P<005). However, the mitochondrial membrane potential was significanly decreased in the pEGFP-Mfn-2 transfection group (\[1.6±0.1\] % vs \[5.0±0.5\] %, P<0.05). Conclusion: pEGFP-Mfn-2 transfection can enhance the sensitivity of T47D cells to parthenolide.
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Project Supported:
Project supported by the Natural Science Foundation of Tianjin（No. 10JCYBJC1150）
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