Adeno-associated virus type 2-mediated gene transfer of a short hairpin-RNA targeting human IGFBP-2 suppresses the proliferation and invasion of MDA-MB-468 cells

Gao,Chao; Zhang,Ru‑Song; Zheng,Nan; Wang,Chen

doi:10.3892/mmr.2018.8434

Adeno-associated virus type 2-mediated gene transfer of a short hairpin-RNA targeting human IGFBP-2 suppresses the proliferation and invasion of MDA-MB-468 cells

Authors:
- Chao Gao
- Ru‑Song Zhang
- Nan Zheng
- Chen Wang
View Affiliations

Affiliations: The Center for Clinical Reproductive Medicine, Jiangsu Hospital, Nanjing, Jiangsu 210029, P.R. China, The Pathology Department, Nanjing General Hospital of Nanjing Military Command, Nanjing, Jiangsu 210002, P.R. China, State Key Laboratory of Chemistry for Life Science & Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, Jiangsu 210093, P.R. China
Published online on: January 16, 2018 https://doi.org/10.3892/mmr.2018.8434
Pages: 4383-4391
Copyright: © Gao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Adeno-associated virus 2 (AAV2) is prepotent in the biological treatment of breast tumor because of its low pathogenicity and immunogenicity. Our previous study demonstrated that insulin‑like growth factor‑binding protein 2 (IGFBP‑2) was highly expressed in patients with breast metastasis. In the present study, the effects of recombinant AAV2 on the growth and metastasis of breast cancer cells were determined in vitro, and in vivo. rAAV2-ZsGreen-shRNA-scramble and rAAV2‑ZsGreen‑shRNA‑hIGFBP‑2 were used to transfect MDA‑MB‑468, and MCF‑10A cells respectively, and observed that these virus could not penetrate the normal human breast epithelia MCF‑10A cell line. To investigate the effect of the recombinant virus on chemotherapeutics, paclitaxel was added to MDA‑MB‑468 cells and it was demonstrated that rAAV2‑ZsGreen‑shRNA‑hIGFBP-2-infected MDA-MB-468 cells were highly chemosensitive to paclitaxel compared with rAAV2‑ZsGreen‑shRNA‑scramble‑injected cells. In addition, it was demonstrated that the invasive ability of rAAV2‑ZsGreen‑shRNA‑hIGFBP‑2‑infected MDA-MB-468 cells was highly impaired compared with the rAAV2‑ZsGreen‑shRNA‑scramble group. In the nude mice xenografts, the rAAV2‑ZsGreen‑shRNA‑hIGFBP‑2 injection inhibited tumor growth and Ki‑67 expression was significantly downregulated compared with the scramble group. Following IGFBP‑2 knockdown using rAAV2-ZsGreen-shRNA-hIGFBP‑2, matrix metalloproteinase‑2 expression was significantly reduced in tumor tissues compared with that in rAAV2‑ZsGreen‑shRNA‑scramble treated tumor tissues. These findings have provided a direction for the application of novel AAV2‑based therapeutics for treating aggressive triple‑negative breast cancer types.

View Figures

View References

1	El Atiq F, Garrouste F, Remacle-Bonnet M, Sastre B and Pommier G: Alterations in serum levels of insulin-like growth factors and insulin-like growth-factor-binding proteins in patients with colorectal cancer. Int J Cancer. 57:491–497. 1994. View Article : Google Scholar : PubMed/NCBI
2	Karasik A, Menczer J, Pariente C and Kanety H: Insulin-like growth factor-I (IGF-I) and IGF-binding protein-2 are increased in cyst fluids of epithelial ovarian cancer. J Clin Endocrinol Metab. 78:271–276. 1994. View Article : Google Scholar : PubMed/NCBI
3	Baron-Hay S, Boyle F, Ferrier A and Scott C: Elevated serum insulin-like growth factor binding protein-2 as a prognostic marker in patients with ovarian cancer. Clin Cancer Res. 10:1796–1806. 2004. View Article : Google Scholar : PubMed/NCBI
4	Lee DY, Kim SJ and Lee YC: Serum insulin-like growth factor (IGF)-I and IGF-binding proteins in lung cancer patients. J Kor Med Sci. 14:401–404. 1999. View Article : Google Scholar
5	Cohen P, Peehl DM, Stamey TA, Wilson KF, Clemmons DR and Rosenfeld RG: Elevated levels of insulin-like growth factor-binding protein-2 in the serum of prostate cancer patients. J Clin Endocrinol Metab. 76:1031–1035. 1993. View Article : Google Scholar : PubMed/NCBI
6	Shariat SF, Lamb DJ, Kattan MW, Nguyen C, Kim J, Beck J, Wheeler TM and Slawin KM: Association of preoperative plasma levels of insulin-like growth factor I and insulin-like growth factor binding proteins-2 and −3 with prostate cancer invasion, progression and metastasis. J Clin Oncol. 20:833–841. 2002. View Article : Google Scholar : PubMed/NCBI
7	Busund LT, Richardsen E, Busund R, Ukkonen T, Bjørnsen T, Busch C and Stalsberg H: Significant expression of IGFBP2 in breast cancer compared with benign lesions. J Clin Pathol. 58:361–366. 2005. View Article : Google Scholar : PubMed/NCBI
8	So AI, Levitt RJ, Eigl B, Fazli L, Muramaki M, Leung S, Cheang MC, Nielsen TO, Gleave M and Pollak M: Insulin-like growth factor binding protein-2 is a novel therapeutic target associated with breast cancer. Clin Cancer Res. 14:6944–6954. 2008. View Article : Google Scholar : PubMed/NCBI
9	Schütt BS, Langkamp M, Rauschnabel U, Ranke MB and Elmlinger MW: Integrin-mediated action of insulin-like growth factor binding protein-2 in tumor cells. J Mol Endocrinol. 32:859–868. 2004. View Article : Google Scholar : PubMed/NCBI
10	Pereira JJ, Meyer T, Docherty SE, Reid HH, Marshall J, Thompson EW, Rossjohn J and Price JT: Bimolecular interaction of insulin-like growth factor (IGF) binding protein-2 with alphavbeta3 negatively modulates IGF-I-mediated migration and tumor growth. Cancer Res. 64:977–984. 2004. View Article : Google Scholar : PubMed/NCBI
11	Wang C, Gao C, Meng K, Qiao H and Wang Y: Human adipocytes stimulate invasion of breast cancer MCF-7 cells by secreting IGFBP-2. PLoS One. 10:e01193482015. View Article : Google Scholar : PubMed/NCBI
12	Zaiss AK and Muruve DA: Immunity to adeno-associated virus vectors in animals and humans: A continued challenge. Gene Ther. 15:808–816. 2008. View Article : Google Scholar : PubMed/NCBI
13	Herzog RW, Yang EY, Couto LB, Hagstrom JN, Elwell D, Fields PA, Burton M, Bellinger DA, Read MS, Brinkhous KM, et al: Long-term correction of canine hemophilia B by gene transfer of blood coagulation factor IX mediated by adeno-associated viral vector. Nat Med. 5:56–63. 1999. View Article : Google Scholar : PubMed/NCBI
14	Wang L, Takabe K, Bidlingmaier SM, Ill CR and Verma IM: Sustained correction of bleeding disorder in hemophilia B mice by gene therapy. Proc Natl Acad Sci USA. 96:pp. 3906–3910. 1999; View Article : Google Scholar : PubMed/NCBI
15	Ponnazhagan S, Mahendra G, Kumar S, Shaw DR, Stockard CR, Grizzle WE and Meleth S: Adeno-associated virus 2-mediated antiangiogenic cancer gene therapy: Long-term efficacy of a vector encoding angiostatin and endostatin over vectors encoding a single factor. Cancer Res. 64:1781–1787. 2004. View Article : Google Scholar : PubMed/NCBI
16	Collins SA, Buhles A, Scallan MF, Harrison PT, O'Hanlon DM, O'Sullivan GC and Tangney M: AAV2-mediated in vivo immune gene therapy of solid tumours. Genet Vaccines Ther. 8:82010. View Article : Google Scholar : PubMed/NCBI
17	Alam S, Bowser BS, Conway MJ, Israr M, Tandon A and Meyers C: Adeno-associated virus type 2 infection activates caspase dependent and independent apoptosis in multiple breast cancer lines but not in normal mammary epithelial cells. Mol Cancer. 10:972011. View Article : Google Scholar : PubMed/NCBI
18	Foulstone EJ, Zeng L, Perks CM and Holly JM: Insulin-like growth factor binding protein 2 (IGFBP-2) promotes growth and survival of breast epithelial cells: Novel regulation of the estrogen receptor. Endocrinology. 154:1780–1793. 2013. View Article : Google Scholar : PubMed/NCBI
19	Wang C, Bian Z, Wei D and Zhang JG: MiR-29b regulates migration of human breast cancer cells. Mol Cell Biochem. 352:197–207. 2011. View Article : Google Scholar : PubMed/NCBI
20	Uzoh CC, Holly JM, Biernacka KM, Persad RA, Bahl A, Gillatt D and Perks CM: Insulin-like growth factor-binding protein-2 promotes prostate cancer cell growth via IGF-dependent or -independent mechanisms and reduces the efficacy of docetaxel. Br J Cancer. 104:1587–1593. 2011. View Article : Google Scholar : PubMed/NCBI
21	Holmes KM, Annala M, Chua CY, Dunlap SM, Liu Y, Hugen N, Moore LM, Cogdell D, Hu L, Nykter M, et al: Insulin-like growth factor-binding protein 2-driven glioma progression is prevented by blocking a clinically significant integrin, integrin-linked kinase, and NF-κB network. Proc Natl Acad Sci USA. 109:pp. 3475–3480. 2012; View Article : Google Scholar : PubMed/NCBI
22	Kiepe D, Van Der A, Ciarmatori S, Ständker L, Schütt B, Hoeflich A, Hügel U, Oh J and Tönshoff B: Defined carboxy-terminal fragments of insulin-like growth factor (IGF) binding protein-2 exert similar mitogenic activity on cultured rat growth plate chondrocytes as IGF-I. Endocrinology. 149:4901–4911. 2008. View Article : Google Scholar : PubMed/NCBI
23	Wang GK, Hu L, Fuller GN and Zhang W: An interaction between insulin-like growth factor-binding protein 2 (IGFBP2) and integrin alpha5 is essential for IGFBP2-induced cell mobility. J Biol Chem. 281:14085–14091. 2006. View Article : Google Scholar : PubMed/NCBI
24	Sehgal P, Kumar N, Praveen Kumar VR, Patil S, Bhattacharya A, Vijaya Kumar M, Mukherjee G and Kondaiah P: Regulation of protumorigenic pathways by insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis. Mol Cancer. 12:632013. View Article : Google Scholar : PubMed/NCBI
25	Chakrabarty S and Kondratick L: Insulin-like growth factor binding protein-2 stimulates proliferation and activates multiple cascades of the mitogen-activated protein kinase pathways in NIH-OVCAR3 human epithelial ovarian cancer cells. Cancer Biol Ther. 5:189–197. 2006. View Article : Google Scholar : PubMed/NCBI
26	Chatterjee S, Park ES and Soloff MS: Proliferation of DU145 prostate cancer cells is inhibited by suppressing insulin-like growth factor binding protein-2. Int J Urol. 11:876–884. 2004. View Article : Google Scholar : PubMed/NCBI
27	Miyake H, Hara I, Yamanaka K, Muramaki M, Gleave M and Eto H: Introduction of insulin-like growth factor binding protein-2 gene into human bladder cancer cells enhances their metastatic potential. Oncol Rep. 13:341–345. 2005.PubMed/NCBI
28	Fukushima T, Tezuka T, Shimomura T, Nakano S and Kataoka H: Silencing of insulin-like growth factor-binding protein-2 in human glioblastoma cells reduces both invasiveness and expression of progression-associated gene CD24. J Biol Chem. 282:18634–18644. 2007. View Article : Google Scholar : PubMed/NCBI
29	Alam S, Bowser BS, Israr M, Conway MJ and Meyers C: Adeno-associated virus type 2 infection of nude mouse human breast cancer xenograft induces necrotic death and inhibits tumor growth. Cancer Biol Ther. 15:1013–1028. 2014. View Article : Google Scholar : PubMed/NCBI