HBX蛋白对正常肝细胞增殖及端粒酶活性的影响

doi:10.3969/j.issn.1004-616x.2012.03.005

›› 2012, Vol. 24 ›› Issue (3): 183-187.doi: 10.3969/j.issn.1004-616x.2012.03.005

HBX蛋白对正常肝细胞增殖及端粒酶活性的影响

贺丹丹,邓志华,杨倩

山西省太原市山西医科大学第二医院消化科，030001

收稿日期:2012-01-17 修回日期:2012-03-22 出版日期:2012-05-30 发布日期:2012-05-30
通讯作者: 邓志华

The effect of HBX protein on the proliferation of normal hepatic cells and on telomerase activity

HE Dan-dan，DENG Zhi-hua，YANG Qian

Department of Gastroenterology, Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China

Received:2012-01-17 Revised:2012-03-22 Online:2012-05-30 Published:2012-05-30
Contact: DENG Zhi-hua

摘要/Abstract

摘要： 目的：观察乙型肝炎病毒X (hepatitis B virus X，HBX)蛋白对L02肝细胞的转化作用及其对端粒酶活性的影响。方法：构建PEGFP-N1-HBX质粒，脂质体介导转染L02细胞，倒置荧光显微镜观察EGFP的表达。稳定转染后Western blotting检测L02细胞中HBX蛋白的表达，电子显微镜观察L02细胞的形态，MTT法检测细胞的增殖，RT-PCR检测L02细胞端粒酶逆转录酶 (hTERT)mRNA的表达，TRAP-PCR银染法检测L02细胞端粒酶活性的变化。结果：成功构建了PEGFP-N1-HBX质粒，稳定转染后L02细胞中有HBX蛋白的表达，且细胞形态明显幼稚化，有恶性变趋势，细胞增殖能力明显增加 (P<0.05)，细胞hTERT mRNA表达明显增加，端粒酶被激活。结论：HBX蛋白可诱导正常肝细胞的恶性转化，上调hTERT mRNA的表达并能激活端粒酶。

关键词: 乙型肝炎病毒X蛋白, 肝肿瘤, 端粒酶, 基因治疗

Abstract: OBJECTIVE: To observe the transformation in normal hepatic cell line L02 by the hepatitis B virus X (HBX) protein and the effect of HBX protein on the activity of telomerase. METHODS：PEGFP-N1-HBX plasmids were constructed and transfected into L02 normal liver cells，the expression of EGFP was assessed by fluorescence microscope. The expression of HBX protein in L02 cells was measured by Western blotting analysis. Cellular morphology was studied by electron microscope. The proliferation of L02 cells after stably transfected with plasmids was measured by MTT assay. The expressions of hTERT mRNA was detected by RT-PCR in L02 cells after stably transfected with plasmids. The activity of telomerase was evaluated by TRAP-PCR argentation in L02 cells after stably transfected with plasmids. RESULTS：PEGFP-N1-HBX plasmid was successfully constructed. The expression of HBX protein was observed in normal liver L02 cells. The cell morphology after transfection with PEGFP-N1-HBX plasmid appeared immature and with the trend of malignant transformation. Compared with the L02 cells after transfection with PEGFP-N1 plasmid，PEGFP-N1-HBX transfection promoted the proliferation of L02 cells (P<0.05).The hTERT mRNA expression increased in L02 cells after transfection with PEGFP-N1-HBX plasmid and telomerase was activated. CONCLUSION： HBX protein could induce normal liver L02 cells torword malignant transformation，enhance the expression of hTERT mRNA and activate telomerase.

Key words: hepatitis B virus X protein, hepatocellular carcinoma, telomerase, gene therapy

中图分类号:

R735.7

贺丹丹,邓志华,杨倩. HBX蛋白对正常肝细胞增殖及端粒酶活性的影响[J]. , 2012, 24(3): 183-187.

HE Dan-dan，DENG Zhi-hua，YANG Qian. The effect of HBX protein on the proliferation of normal hepatic cells and on telomerase activity[J]. , 2012, 24(3): 183-187.

[1]	肖月, 李格非, 刘丹, 陈婷, 林桂淼. 多功能非病毒载体在肿瘤基因治疗中的应用[J]. 癌变·畸变·突变, 2019, 31(3): 249-252,256.
[2]	何洁, 万经海, 李学记, 钱海鹏, 孟肖利, 郝佳洁, 徐昕, 王明荣. 胶质瘤中端粒酶逆转录酶启动子区突变分析及其预后意义[J]. 癌变·畸变·突变, 2015, 27(5): 361-365.
[3]	梁朝辉(综述), 焦保华*(审校). 以Wip1为靶点的抗肿瘤研究进展[J]. , 2011, 23(4): 312-314.
[4]	孙国贵1(综述)，王雅棣2,*，郑明民2(审校). 锰超氧化物歧化酶与肿瘤[J]. , 2011, 23(2): 158-.
[5]	陈进军1，赵　路2，董　玫3,王思明4，秦琪瑜1，陈　虎1，史清文3,. 土木香根中5种倍半萜化合物抗肝癌活性的研究[J]. , 2010, 22(6): 440-443.
[6]	周建农1,马国建2,何晓松2,沈宗丽2,薛开先2,. 循环肿瘤细胞p53基因表达和端粒酶活性在大肠癌临床病理学中的意义[J]. , 2009, 21(2): 145-148.
[7]	卓志红(综述)，龚朝辉(审校). DNA甲基化与乳腺癌[J]. , 2008, 20(2): 163-165.
[8]	高　敏 /周福祥 /谢丛华 /肖创映 /周云峰. 端粒酶抑制剂叠氮胸苷对HeLa细胞放射性DNA损伤修复的影响[J]. , 2007, 19(6): 448-452.
[9]	何迎春(综述)/ 田道法(审校). 基因表达异常与癌前病变演化[J]. , 2007, 19(5): 421-423.
[10]	李　军张爱华岑笃才黄晓欣蒋宪瑶. 砷中毒患者病变皮肤中端粒酶活性与PCNA表达及其意义[J]. , 2007, 19(4): 276-279.
[11]	陈琳琳;朱玉萍;宋章法;黄学锋;毛伟芳. 结肠癌TRAIL耐药细胞株的建立及其耐药机制的初步研究[J]. , 2006, 18(5): 340-343.
[12]	王　丽;陈治文;夏　俊. 三氧化二砷抑制恶性黑色素瘤B16细胞生长及对端粒酶活性的影响[J]. , 2005, 17(5): 287-290.
[13]	邢　岩/综述;田庆伟/审校. 端粒酶与P53在细胞癌变中关系的研究进展[J]. , 2005, 17(1): 62-64.
[14]	张　敏;奚耕思;戴　璐. 端粒酶与癌症的关系研究进展[J]. , 2005, 17(1): 59-61.
[15]	张红宇，孙　强，王亚东，张维祥，杨卫军，张锦瑜. 野生型P53、P16基因联合抑制人胃癌细胞HGC27生长的实验研究[J]. , 2004, 16(6): 344-346.

HBX蛋白对正常肝细胞增殖及端粒酶活性的影响

The effect of HBX protein on the proliferation of normal hepatic cells and on telomerase activity

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价