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The Overexpression of PHP14 and Its Effect on NIH-3T3 Cells Proliferation in vitro and Anchor Independent Growth |
XU An-jian1,2, CUI Yong3, ZHAO Gao-chao2, WU Shan-na2, LÜ Zhi4, JIN Qing-e2, GU Jun-chao1 |
1. Beijing Friendship Hospital, Beijing 100050, China;
2. Viral Laboratory, Beijing Friendship Hospital, Beijing 100050, China;
3. Department of Chest Surgery, Beijing Friendship Hospital, Beijing 100050, China;
4. Department of Clinical Laboratories, Beijing Friendship Hospital, Beijing 100050, China |
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Abstract Objective: To amplify human PHP14 gene, and construct the N-terminal and C-terminal GFP fused vectors, establish the overexpress PHP14 NIH-3T3 cell line to investigate its effect on NIH-3T3 cell proliferation and anchor independent growth. Methods: The cDNA sequence of PHP14 gene was amplified and sub-cloned into pEGFP-N and pEGFP-C3 vectors. Transfecting of those vectors into NIH-3T3 cells and investigating the cell proliferation and anchor independent growth in PHP14 overexpressed NIH-3T3 cells using MTT and Soft agar colony assay. Results: The pEGFP-N2-PHP14 and pEGFP-C3-PHP14 prokaryotic expression vectors were constructed successfully and the expression of those vectors were detected in NIH-3T3 cells. Overexpression of PHP14 in NIH-3T3 cells did not affect the cell proliferation of NIH-3T3 cells, but gave the NIH-3T3 cells the ability of anchor independent growth. Conclusion: PHP14 prokaryotic expression vectors were constructed successfully and overexpression of PHP14 in NIH-3T3 cells did not affect the cell proliferation of NIH-3T3 cells, but gave the NIH-3T3 cells the ability of anchor independent growth.
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Received: 15 February 2013
Published: 25 June 2013
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