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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2013, Vol. 33 Issue (6): 7-11    
研究报告     
人组氨酸磷酸酶蛋白PHP14的过表达及其对NIH-3T3细胞体外增殖和非锚定依赖性生长的影响
徐安健1,2, 崔永3, 赵高潮2, 乌姗娜2, 吕志4, 靳庆娥2, 谷俊朝1
1. 首都医科大学附属北京友谊医院 北京 100010;
2. 首都医科大学附属北京友谊医院病毒室 北京 100010;
3. 首都医科大学附属北京友谊医院胸外科 北京 100010;
4. 首都医科大学附属北京友谊医院检验科 北京 100010
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
 全文: PDF(626 KB)   HTML
摘要：

目的:获取人组氨酸磷酸酶蛋白PHP14基因,并构建其N端和C端GFP融合的真核表达载体,建立过表达PHP14的NIH-3T3细胞系,并观察其对NIH-3T3细胞体外增殖和非锚定依赖性生长的影响。方法:以HeLa cDNA为模板,PCR扩增PHP14的全长编码基因,分别克隆到pEGFP-N2和pEGFP-C3载体中,构建pEGFP-N2-PHP14和pEGFP-C3-PHP14真核表达载体,利用脂质体将构建的载体转染到NIH-3T3细胞中,MTT法检测细胞增殖,软琼脂成集落法检测体外非锚定依赖性生长能力。结果:成功构建了过表达PHP14的真核表达载体pEGFP-N2-PHP14和pEGFP-C3-PHP14,并在NIH-3T3细胞中检测到了目的基因的过表达,PHP14在NIH-3T3细胞中过表达并不影响NIH-3T3细胞的体外增殖,但赋予了NIH-3T3细胞非锚定依赖性生长的能力。结论:成功构建了过表达PHP14的NIH-3T3细胞模型,在NIH-3T3细胞中过表达PHP14并不影响NIH-3T3细胞的体外增殖,但赋予了NIH-3T3细胞非锚定依赖性生长能力。
关键词： PHP14;  过表达;  增殖;  非锚定依赖性生长    
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.
Key words: PHP14    Overexpression    Cell proliferation    Anchor independent growth
收稿日期: 2013-02-15 出版日期: 2013-06-25
ZTFLH:  Q819  
通讯作者: 徐安健     E-mail: xuanjian1981@yahoo.com.cn
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引用本文:

徐安健, 崔永, 赵高潮, 乌姗娜, 吕志, 靳庆娥, 谷俊朝. 人组氨酸磷酸酶蛋白PHP14的过表达及其对NIH-3T3细胞体外增殖和非锚定依赖性生长的影响[J]. 中国生物工程杂志, 2013, 33(6): 7-11.

XU An-jian, CUI Yong, ZHAO Gao-chao, WU Shan-na, LÜ Zhi, JIN Qing-e, GU Jun-chao. The Overexpression of PHP14 and Its Effect on NIH-3T3 Cells Proliferation in vitro and Anchor Independent Growth. China Biotechnology, 2013, 33(6): 7-11.

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https://manu60.magtech.com.cn/biotech/CN/    或    https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I6/7

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