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| Construction of recombinant expression lentivirus vector carrying melanotransferrin and identification of its role in cervical cancer HeLa cells |
| YANG Yue, LI Wan, LU Chun |
| (Department of Microbiology, Nanjing Medical University, Nanjing Jiangsu 211166, China) |
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Abstract Objective: To construct the recombinant lentivirus vector carrying melanotransferrin (MFI2) for upregulating MFI2 expression level in HeLa cells and investigating its effect on HeLa cells proliferation. Methods: The sequences of MFI2 were cloned from cDNA of HeLa cells and were inserted into lentiviral plasmid pHAGE-CMV-MCS-IzsGreen(pHAGE) to construct the recombinant plasmid pHAGE-MFI2, followed by cotransfection of the package plasmid psPAX2 and the envelope plasmid pMD2. G into 293T cells. The titer of the generated MFI2-expressing lentivirus was determined by gradient dilution. The MFI2-expressing lentivirus was transduced into the HeLa cells and the expression of MFI2 was determined by Western blotting, and then the proliferation of the treated HeLa cells was further studied via CCK-8 assay and wound healing. Results: Both the identification of plasmid double enzyme digestion and nucleic acid sequencing comparison results confirmed the successful construction of pHAGE-MIF2 plasmid.MFI2 gene fusion lentivirus was successfully constructed by lentivirus triplasmid collaborative packaging system, and the virus titer was up to 6×107 TU/mL.After the infection of HeLa cells with MFI2 lentivirus, the expression level of MFI2 was significantly increased, while the cell proliferation activity was significantly decreased. Conclusion: The overexpression of MFI2 could inhibit the proliferation of HeLa cells with high efficiency.
［Key words］melanotransferrin; cervical cancer; lentivirus; cell proliferation
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Received: 22 January 2019
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2019, Vol. 29 
