Magnetic gold nanoparticle-mediated small interference RNA silencing Bag-1 gene for colon cancer therapy

Huang,Wenbai; Liu,Zhan'ao; Zhou,Guanzhou; Tian,Ailing; Sun,Nianfeng

doi:10.3892/or.2015.4453

Magnetic gold nanoparticle-mediated small interference RNA silencing Bag-1 gene for colon cancer therapy

Authors:
- Wenbai Huang
- Zhan'ao Liu
- Guanzhou Zhou
- Ailing Tian
- Nianfeng Sun
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Affiliations: Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 200012, P.R. China
Published online on: November 26, 2015 https://doi.org/10.3892/or.2015.4453
Pages: 978-984

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Abstract

Bcl-2-associated athanogene 1 (Bag-1) is a positive regulator of Bcl-2 which is an anti-apoptotic gene. Bag-1 was very slightly expressed in normal tissues, but often highly expressed in many tumor tissues, particularly in colon cancer, which can promote metastasis, poor prognosis and anti-apoptotic function of colon cancer. We prepared and evaluated magnetic gold nanoparticle/Bag-1 siRNA recombinant plasmid complex, a gene therapy system, which can transfect cells efficiently, for both therapeutic effect and safety in vitro mainly by electrophoretic mobility shift assays, flow cytometric analyses, cell viability assays, western blot analyses and RT-PCR (real-time) assays. Magnetic gold nanoparticle/Bag-1 siRNA recombinant plasmid complex was successfully transfected into LoVo colon cancer cells and the exogenous gene was expressed in the cells. Flow cytometric results showed apoptosis rate was significantly increased. In MTT assays, magnetic gold nanoparticles revealed lower cytotoxicity than Lipofectamine 2000 transfection reagents (P<0.05). Both in western blot analyses and RT-PCR assays, magnetic gold nanoparticle/Bag-1 siRNA recombinant plasmid complex transfected cells demonstrated expression of Bag-1 mRNA (P<0.05) and protein (P<0.05) was decreased. In further study, c-myc and β-catenin which are main molecules of Wnt/β‑catenin pathway were decreased when Bag-1 were silenced in nanoparticle plasmid complex transfected LoVo cells. These results suggest that magnetic gold nanoparticle mediated siRNA silencing Bag-1 is an effective gene therapy method for colon cancer.

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1	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
2	Qiu K, Yu B, Huang H, Zhang P, Huang J, Zou S, Chen Y, Ji L and Chao H: A dendritic nano-sized hexanuclear ruthenium(II) complex as a one- and two-photon luminescent tracking non-viral gene vector. Sci Rep. 5:107072015. View Article : Google Scholar : PubMed/NCBI
3	Şalva E, Turan SO, Ekentok C and Akbuğa J: Generation of stable cell line by using chitosan as gene delivery system. Cytotechnology. Jul 2–2015.Epub ahead of print. View Article : Google Scholar
4	Hassan M, Watari H, AbuAlmaaty A, Ohba Y and Sakuragi N: Apoptosis and molecular targeting therapy in cancer. Biomed Res Int. 2014:1508452014. View Article : Google Scholar : PubMed/NCBI
5	van der Zee JA, Ten Hagen TL, Hop WC, van Dekken H, Dicheva BM, Seynhaeve AL, Koning GA, Eggermont AM and van Eijck CH: Bcl-2 associated anthanogen-1 (Bag-1) expression and prognostic value in pancreatic head and periampullary cancer. Eur J Cancer. 49:323–328. 2013. View Article : Google Scholar
6	Sun N, Meng Q and Tian A: Expressions of the anti-apoptotic genes Bag-1 and Bcl-2 in colon cancer and their relationship. Am J Surg. 200:341–345. 2010. View Article : Google Scholar : PubMed/NCBI
7	Clemo NK, Collard TJ, Southern SL, Edwards KD, Moorghen M, Packham G, Hague A, Paraskeva C and Williams AC: BAG-1 is up-regulated in colorectal tumour progression and promotes colorectal tumour cell survival through increased NF-kappaB activity. Carcinogenesis. 29:849–857. 2008. View Article : Google Scholar : PubMed/NCBI
8	Kievit FM and Zhang M: Surface engineering of iron oxide nanoparticles for targeted cancer therapy. Acc Chem Res. 44:853–862. 2011. View Article : Google Scholar : PubMed/NCBI
9	Liu M, Wang Z, Zong S, Chen H, Zhu D, Zhong Y and Cui Y: Remote-controlled DNA release from Fe₃O₄ @Au nanoparticles using an alternating electromagnetic field. J Biomed Nanotechnol. 11:979–987. 2015. View Article : Google Scholar : PubMed/NCBI
10	Raji MA, Amara M, Amoabediny G, Tajik P, Barin A, Magierowski S and Ghafar-Zadeh E: Cytotoxicity of synthesized iron oxide nanoparticles: Toward novel biomarkers of colon cancer. Conf Proc IEEE Eng Med Biol Soc. 2014:6179–6182. 2014.
11	Wan Q, Xie L, Gao L, Wang Z, Nan X, Lei H, Long X, Chen ZY, He CY, Liu G, et al: Self-assembled magnetic theranostic nanoparticles for highly sensitive MRI of minicircle DNA delivery. Nanoscale. 5:744–752. 2013. View Article : Google Scholar
12	Sun NF, Liu ZA, Huang WB, Tian AL and Hu SY: The research of nanoparticles as gene vector for tumor gene therapy. Crit Rev Oncol Hematol. 89:352–357. 2014. View Article : Google Scholar
13	Singh D, McMillan JM, Kabanov AV, Sokolsky-Papkov M and Gendelman HE: Bench-to-bedside translation of magnetic nanoparticles. Nanomedicine. 9:501–516. 2014. View Article : Google Scholar : PubMed/NCBI
14	Ben Q, An W, Fei J, Xu M, Li G, Li Z and Yuan Y: Downregulation of L1CAM inhibits proliferation, invasion and arrests cell cycle progression in pancreatic cancer cells in vitro. Exp Ther Med. 7:785–790. 2014.PubMed/NCBI
15	Zhou Y, Xiong M, Niu J, Sun Q, Su W, Zen K, Dai C and Yang J: Secreted fibroblast-derived miR-34a induces tubular cell apoptosis in fibrotic kidney. J Cell Sci. 127:4494–4506. 2014. View Article : Google Scholar : PubMed/NCBI
16	Maddalo D, Neeb A, Jehle K, Schmitz K, Muhle-Goll C, Shatkina L, Walther TV, Bruchmann A, Gopal SM, Wenzel W, et al: A peptidic unconjugated GRP78/BiP ligand modulates the unfolded protein response and induces prostate cancer cell death. PLoS One. 7:e456902012. View Article : Google Scholar : PubMed/NCBI
17	Roth W, Grimmel C, Rieger L, Strik H, Takayama S, Krajewski S, Meyermann R, Dichgans J, Reed JC and Weller M: Bag-1 and Bcl-2 gene transfer in malignant glioma: Modulation of cell cycle regulation and apoptosis. Brain Pathol. 10:223–234. 2000. View Article : Google Scholar : PubMed/NCBI
18	Wood J, Lee SS and Hague A: Bag-1 proteins in oral squamous cell carcinoma. Oral Oncol. 45:94–102. 2009. View Article : Google Scholar
19	Aust S, Pils S, Polterauer S, Horvat R, Cacsire Castillo-Tong D, Pils D, Dudek G, Schmid B, Speiser P, Reinthaller A, et al: Expression of Bcl-2 and the antiapoptotic BAG family proteins in ovarian cancer. Appl Immunohistochem Mol Morphol. 21:518–524. 2013. View Article : Google Scholar : PubMed/NCBI
20	Liu H, Lu S, Gu L, Gao Y, Wang T, Zhao J, Rao J, Chen J, Hao X and Tang SC: Modulation of BAG-1 expression alters the sensitivity of breast cancer cells to tamoxifen. Cell Physiol Biochem. 33:365–374. 2014. View Article : Google Scholar : PubMed/NCBI
21	Mashukova A, Kozhekbaeva Z, Forteza R, Dulam V, Figueroa Y, Warren R and Salas PJ: The BAG-1 isoform BAG-1M regulates keratin-associated Hsp70 chaperoning of aPKC in intestinal cells during activation of inflammatory signaling. J Cell Sci. 127:3568–3577. 2014. View Article : Google Scholar : PubMed/NCBI
22	Li P, Wang YD, Cheng J, Chen JC and Ha MW: Association between polymorphisms of BAG-1 and XPD and chemotherapy sensitivity in advanced non-small-cell lung cancer patients treated with vinorelbine combined cisplatin regimen. Tumour Biol. Jun 30–2015.Epub ahead of print.
23	Bai YX, Yi JL, Li JF and Sui H: Clinicopathologic significance of BAG1 and TIMP3 expression in colon carcinoma. World J Gastroenterol. 13:3883–3885. 2007. View Article : Google Scholar : PubMed/NCBI
24	Knee DA, Froesch BA, Nuber U, Takayama S and Reed JC: Structure-function analysis of Bag1 proteins. Effects on androgen receptor transcriptional activity. J Biol Chem. 276:12718–12724. 2001. View Article : Google Scholar : PubMed/NCBI
25	Schmidt U, Holsboer F and Rein T: Role of the hsp70 cochaperone BAG1 in glucocorticoid receptor function and stress-related diseases. Proc Natl Acad Sci USA. 105:E101author reply E102. 2008. View Article : Google Scholar : PubMed/NCBI
26	Brendel A, Felzen V, Morawe T, Manthey D and Behl C: Differential regulation of apoptosis-associated genes by estrogen receptor alpha in human neuroblastoma cells. Restor Neurol Neurosci. 31:199–211. 2013.
27	Brive L, Takayama S, Briknarová K, Homma S, Ishida SK, Reed JC and Ely KR: The carboxyl-terminal lobe of Hsc70 ATPase domain is sufficient for binding to BAG1. Biochem Biophys Res Commun. 289:1099–1105. 2001. View Article : Google Scholar : PubMed/NCBI
28	Rauch JN and Gestwicki JE: Binding of human nucleotide exchange factors to heat shock protein 70 (Hsp70) generates functionally distinct complexes in vitro. J Biol Chem. 289:1402–1414. 2014. View Article : Google Scholar :
29	Jang GB, Kim JY, Cho SD, Park KS, Jung JY, Lee HY, Hong IS and Nam JS: Blockade of Wnt/β-catenin signaling suppresses breast cancer metastasis by inhibiting CSC-like phenotype. Sci Rep. 5:124652015. View Article : Google Scholar
30	Li F, Wang T and Tang S: SOX14 promotes proliferation and invasion of cervical cancer cells through Wnt/β-catenin pathway. Int J Clin Exp Pathol. 8:1698–1704. 2015.
31	Akaboshi S, Watanabe S, Hino Y, Sekita Y, Xi Y, Araki K, Yamamura K, Oshima M, Ito T, Baba H, et al: HMGA1 is induced by Wnt/beta-catenin pathway and maintains cell proliferation in gastric cancer. Am J Pathol. 175:1675–1685. 2009. View Article : Google Scholar : PubMed/NCBI
32	Wang Y, He L, Du Y, Zhu P, Huang G, Luo J, Yan X, Ye B, Li C, Xia P, et al: The long noncoding RNA lncTCF7 promotes self-renewal of human liver cancer stem cells through activation of Wnt signaling. Cell Stem Cell. 16:413–425. 2015. View Article : Google Scholar : PubMed/NCBI
33	Atkinson JM, Rank KB, Zeng Y, Capen A, Yadav V, Manro JR, Engler TA and Chedid M: Activating the Wnt/β-Catenin pathway for the treatment of melanoma - Application of LY2090314, a novel selective inhibitor of glycogen synthase kinase-3. PLoS One. 10:e01250282015. View Article : Google Scholar
34	Hu T, Xie N, Qin C, Wang J and You Y: Glucose-regulated protein 94 is a novel glioma biomarker and promotes the aggressiveness of glioma via Wnt/β-catenin signaling pathway. Tumour Biol. Jun 25–2015.Epub ahead of print. View Article : Google Scholar
35	Cai J, Maitra A, Anders RA, Taketo MM and Pan D: β-Catenin destruction complex-independent regulation of Hippo-YAP signaling by APC in intestinal tumorigenesis. Genes Dev. 29:1493–1506. 2015. View Article : Google Scholar : PubMed/NCBI
36	Mir R, Pradhan SJ, Patil P, Mulherkar R and Galande S: Wnt/β-catenin signaling regulated SATB1 promotes colorectal cancer tumorigenesis and progression. Oncogene. Jul 13–2015.Epub ahead of print. View Article : Google Scholar
37	Zhao R, Peng X, Chu H and Song W: Phosphorylatable short peptide conjugated chitosan mediated gene therapy for repair of articular cartilage defect in rabbits. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 28:1346–1352. 2014.In Chinese.
38	Son S, Namgung R, Kim J, Singha K and Kim WJ: Bioreducible polymers for gene silencing and delivery. Acc Chem Res. 45:1100–1112. 2012. View Article : Google Scholar
39	Whitehouse A: Herpesvirus saimiri: A potential gene delivery vector (Review). Int J Mol Med. 11:139–148. 2003.PubMed/NCBI
40	Vijayanathan V, Thomas T and Thomas TJ: DNA nanoparticles and development of DNA delivery vehicles for gene therapy. Biochemistry. 41:14085–14094. 2002. View Article : Google Scholar : PubMed/NCBI
41	Jingting C, Huining L and Yi Z: Preparation and characterization of magnetic nanoparticles containing Fe₃O₄-dextran-anti-β-human chorionic gonadotropin, a new generation choriocarcinomaspecific gene vector. Int J Nanomed. 6:285–294. 2011.