细胞穿膜肽VP22增强抑癌蛋白PTEN在BT549细胞的表达和分布

余娴, 雷军, 胥正敏, 李婷婷

中国药学杂志 ›› 2015, Vol. 50 ›› Issue (11) : 960-964.

PDF(843 KB)
中国药学杂志
PDF(843 KB)
中国药学杂志 ›› 2015, Vol. 50 ›› Issue (11) : 960-964. DOI: 10.11669/cpj.2015.11.011
·论 著·

细胞穿膜肽VP22增强抑癌蛋白PTEN在BT549细胞的表达和分布

  • 余娴1, 雷军2 , 胥正敏2, 李婷婷2
作者信息 +

VP22 Enhances the Expression and Distribution of the Tumor Suppressor Protein PTEN in BT549

  • YU Xian1, LEI Jun2, XU Zheng-min2, LI Ting-ting2
Author information +
文章历史 +

摘要

目的 制备表达细胞穿膜肽融合蛋白PTEN-VP22 的真核表达质粒,验证细胞穿膜肽 VP22能否增强抑癌蛋白PTEN在BT549细胞的表达和分布。方法 构建重组真核表达质粒pcDNA3-PTEN与pcDNA3-PTEN-VP22,转染BT549细胞,Western blot与免疫荧光检测PTEN蛋白与PTEN-VP22融合蛋白的表达与分布。结果 PTEN蛋白与PTEN-VP22融合蛋白在BT549细胞中成功表达;PTEN-VP22在转染细胞中的表达与分布较之PTEN明显增多,且在细胞间的分布呈现典型的VP22分布特点,并主要分布于细胞核。结论 细胞穿膜肽VP22增强了抑癌蛋白PTEN在BT549细胞的表达和分布。

Abstract

OBJECTIVE To prepare a eukaryotic expression vector for cell penetrating peptide fusion protein PTEN-VP22, and to evaluate whether the cell penetrating peptide VP22 increase the expression and distribution of the tumor suppressor protein PTEN in BT549. METHODS The eukaryotic expression vectors pcDNA3-PTEN and pcDNA3-PTEN-VP22 were constructed and then transfected to BT549. The expression and distribution of PTEN protein and PTEN-VP22 fusion protein were identified by Western blot and immunofluorescence. RESULTS PTEN protein and PTEN-VP22 fusion protein were successfully expressed in BT549. The expression and distribution of PTEN-VP22 were significantly increased compared with those of PTEN. The distribution of PTEN-VP22 was in a typical VP22 pattern and mainly distributed in nucleus. CONCLUSION The cell penetrating peptide VP22 enhances the expression and distribution of the tumor suppressor protein PTEN in BT549.

关键词

PTEN / VP22 / BT549 / 细胞穿膜肽 / 基因治疗

Key words

PTEN / VP22 / BT549 / cell penetrating peptides / gene therapy

引用本文

EndNote

Ris (Procite)

Bibtex

导出引用
余娴, 雷军, 胥正敏, 李婷婷. 细胞穿膜肽VP22增强抑癌蛋白PTEN在BT549细胞的表达和分布[J]. 中国药学杂志, 2015, 50(11): 960-964 https://doi.org/10.11669/cpj.2015.11.011
YU Xian, LEI Jun, XU Zheng-min, LI Ting-ting. VP22 Enhances the Expression and Distribution of the Tumor Suppressor Protein PTEN in BT549[J]. Chinese Pharmaceutical Journal, 2015, 50(11): 960-964 https://doi.org/10.11669/cpj.2015.11.011
中图分类号： R965   

参考文献

[1] VERMA I M, SOMIA N. Gene therapy-promises, problems and prospects.Nature, 1997, 389(6648): 239-242.
[2] LI J, YEN C, LIAW D, et al. PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science,1997,275(5308):1943-1947.
[3] HEITZ F, MORRIS M C, DIVITA G. Twenty years of cell-penetrating peptides: From molecular mechanisms to therapeutics . Br J Pharmacol, 2009, 157(2): 195-206.
[4] WANG F, WANG Y, ZHANG X, et al. Recent progress of cell-penetrating peptides as new carriers for intracellular cargo delivery . J Controlled Release, 2014, 174(1):126-136.
[5] OLSON E S, JIANG T, AGUILERA T A, et al. Activatable cell penetrating peptides linked to nanoparticles as dual probes for in vivo fluorescence and MR imaging of proteases.Proc Natl Acad Sci USA, 2010, 107(9): 4311-4316.
[6] ELLIOTT G, O′HARE P. Intercellular trafficking and protein delivery by a herpesvirus structural protein.Cell,1997,88(2):223-233.
[7] NISHI K, SAIGO K. Cellular internalization of green fluorescent protein fused with herpes simplex virus protein VP22 via a lipid raft-mediated endocytic pathway independent of caveolae and Rho family GTPases but dependent on dynamin and Arf6 . J Biol Chem, 2007, 282(37):27503-27517.
[8] YU X , LIU L, WU L, et al. Herpes simplex virus type 1 tegument protein VP22 is capable of modulating the transcription of viral TK and gC genes via interaction with viral ICP0 . Biochimie, 2010,92(8):1024-1030.
[9] ZAVAGLIA D, FAVROT M C, EYMIN B, et al. Intercellular trafficking and enhanced in vivo antitumour activity of a non-virally delivered P27-VP22 fusion protein . Gene Ther, 2003, 10(4): 314-325.
SHERIDAN P J, LAWRIE A, CROSSMAN D C, et al. VP22-mediated intercellular transport correlates with enhanced biological activity of MybEngrailed but not (HSV-I) thymidine kinase fusion proteins in primary vascular cells following non-viral transfection . J Gene Med, 2005, 7(3): 375-385.
DING L, CHEN S, LIU P, et al. CBP loss cooperates with PTEN haploinsufficiency to drive prostate cancer: Implications for epigenetic therapy. Cancer Res, 2014, 74(7):2050-2061.
GONZALEZ-ANGULO A M, FERRER-LOZANO J, STEMKE-HALE K, et al. PI3K pathway mutations and PTEN levels in primary and metastatic breast cancer . Mol Cancer Ther, 2011, 10(6): 1093-1101.
CUMBERBATCH M, TANG X, BERAN G, et al. Identification of a subset of human non-small cell lung cancer patients with high PI3Kβ and low PTEN expression, more prevalent in squamous cell carcinoma. Clin Cancer Res, 2014, 20(3): 595-603.
SQUARIZE C H, CASTILHO R M, ABRAHAO A C, et al. PTEN deficiency contributes to the development and progression of head and neck cancer . Neoplasia, 2013,15(5): 461-471.
CHEN H L, CHEN Z Y. Expression of survivin and phosphatase and tensin homolog deleted on chromosome ten and their correlations in hepatocellular carcinoma . Chin J Exp Surg(中华实验外科杂志), 2013,30(6):1283-1285.
DING M, CAO X, XU H N, et al. Prostate cancer-specific and potent antitumor effect of a DD3-controlled oncolytic virus harboring the PTEN gene . PloS One, 2012,7(4):e35153.
ZHOU Y A, ZHANG T, ZHAO J B, et al. The adenovirus-mediated transfer of PTEN inhibits the growth of esophageal cancer cells in vitro and in vivo.Biosci Biotechnol Biochem, 2010,74(4):736-740.
TAKEI Y, SAGA Y, MIZUKAMI H, et al. Overexpression of PTEN in ovarian cancer cells suppresses ip dissemination and extends survival in mice . Mol Cancer Ther, 2008, 7(3):704-711.
NISHIKAWA M, OTSUKI T, OTA A, et al. Induction of tumor-specific immune response by gene transfer of Hsp70-cell-penetrating peptide fusion protein to tumors in mice. Mol Ther, 2010,18 (2): 421-428.
JIN G, ZHOU Y, CHAI Q, et al. VP22 and cytosine deaminase fusion gene modified tissue-engineered neural stem cells for glioma therapy . J Cancer Res Clin Oncol, 2013, 139(3):475-483.
AINTS A, GVEN H, GAHRTON G, et al. Mapping of herpes simplex virus-1 VP22 functional domains for inter-and subcellular protein targeting. Gene Ther, 2001, 8(14):1051-1056.

基金

国家自然科学基金资助项目(81102288)
PDF(843 KB)

Accesses

Citation

Detail
段落导航
相关文章

/

〈 〉