ADAM17-siRNA inhibits MCF-7 breast cancer through EGFR-PI3K-AKT activation

Meng,Xiangchao; Hu,Baoshan; Hossain,Mohammad Monir; Chen,Guofu; Sun,Ying; Zhang,Xuepeng

doi:10.3892/ijo.2016.3536

ADAM17-siRNA inhibits MCF-7 breast cancer through EGFR-PI3K-AKT activation

Authors:
- Xiangchao Meng
- Baoshan Hu
- Mohammad Monir Hossain
- Guofu Chen
- Ying Sun
- Xuepeng Zhang
View Affiliations

Affiliations: Department of Surgical Oncology, Affiliated Hospital, North China University of Science and Technology, Tangshan, Hebei 063000, P.R. China, International Education College, North China University of Science and Technology, Tangshan, Hebei 063000, P.R. China, Department of Pathology, Basic Scientific Medical College, North China University of Science and Technology, Tangshan, Hebei 063000, P.R. China
Published online on: May 24, 2016 https://doi.org/10.3892/ijo.2016.3536
Pages: 682-690

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

A disintegrin and metalloproteinase-17 (ADAM17) can cut and release a wide variety of epidermal growth factor receptor (EGFR) ligands to promote survival, invasion and proliferation of cancer cell, and therefore, is considered to be a potential therapeutic target for cancer. The main goal of the present study was to observe the effects of ADAM17 small interfering RNA (ADAM17-siRNA) on human MCF-7 breast cancer and investigate its activation pathway. In vitro, MCF-7 cells were divided into ADAM17-siRNA groups, nonsense siRNA groups, AG1478 (selective EGFR blocker) groups, LY294002 [phosphatidylinositol 3-kinase (PI3K) phosphorylation inhibitor] groups, PD0325901 [mitogen extracellular kinase (MEK) inhibitor] groups and control groups. In vivo, MCF-7 cells were implanted subcutaneously into nude mice and then these mice were randomly divided into ADAM17-siRNA groups, vector groups and control groups. Our data showed that compared with the control groups, ADAM17-siRNA, AG1478 and LY294002 could inhibit the migration and proliferation of MCF-7 cells, but PD0325901 and nonsense siRNA did not show this effect. Except that specific ADAM17-siRNA could inhibit the expression of ADAM17 mRNA, others did not change it. Western blot analysis further confirmed that EGFR-PI3K-AKT signaling pathway is involved in ADAM17-siRNA inhibiting migration and proliferation of MCF-7 cells. Similarly to the former, the growth of MCF-7 breast cancer in nude mice was significantly inhibited by ADAM17-siRNA. Compared with the control group and the vector group, the tumor volume was smaller in the ADAM17-siRNA group, the tissues developed large areas of necrosis, immunohistochemistry showed low expressions of ADAM17 and Ki-67 and western blot analysis proved that the expression of ADAM17 protein in the tissue was also reduced. The present study suggests that ADAM17-siRNA inhibits MCF-7 breast cancer and is activated through the EGFR-PI3K-AKT signaling pathway.

View Figures

View References

1	Krieger N, Bassett MT and Gomez SL: Breast and cervical cancer in 187 countries between 1980 and 2010. Lancet. 379:1391–1392. 2012. View Article : Google Scholar
2	Caiazza F, McGowan PM, Mullooly M, Murray A, Synnott N, O'Donovan N, Flanagan L, Tape CJ, Murphy G, Crown J, et al: Targeting ADAM-17 with an inhibitory monoclonal antibody has antitumour effects in triple-negative breast cancer cells. Br J Cancer. 112:1895–1903. 2015. View Article : Google Scholar : PubMed/NCBI
3	Ocaña A, Amir E, Seruga B, Martin M and Pandiella A: The evolving landscape of protein kinases in breast cancer: Clinical implications. Cancer Treat Rev. 39:68–76. 2013. View Article : Google Scholar
4	Zhang P, Shen M, Fernandez-Patron C and Kassiri Z: ADAMs family and relatives in cardiovascular physiology and pathology. J Mol Cell Cardiol. 93:186–199. 2015. View Article : Google Scholar : PubMed/NCBI
5	Bolger JC and Young LS: ADAM22 as a prognostic and therapeutic drug target in the treatment of endocrine-resistant breast cancer. Vitam Horm. 93:307–321. 2013. View Article : Google Scholar
6	Rose-John S: ADAM17, shedding, TACE as therapeutic targets. Pharmacol Res. 71:19–22. 2013. View Article : Google Scholar : PubMed/NCBI
7	Lu Y, Jiang F, Zheng X, Katakowski M, Buller B, To SS and Chopp M: TGF-β1 promotes motility and invasiveness of glioma cells through activation of ADAM17. Oncol Rep. 25:1329–1335. 2011.PubMed/NCBI
8	Santiago-Josefat B, Esselens C, Bech-Serra JJ and Arribas J: Post-transcriptional up-regulation of ADAM17 upon epidermal growth factor receptor activation and in breast tumors. J Biol Chem. 282:8325–8331. 2007. View Article : Google Scholar : PubMed/NCBI
9	Murphy G: The ADAMs: Signalling scissors in the tumour microenvironment. Nat Rev Cancer. 8:929–941. 2008. View Article : Google Scholar : PubMed/NCBI
10	Rego SL, Helms RS and Dréau D: Tumor necrosis factor-alpha-converting enzyme activities and tumor-associated macrophages in breast cancer. Immunol Res. 58:87–100. 2014. View Article : Google Scholar
11	Maretzky T, Zhou W, Huang XY and Blobel CP: A transforming Src mutant increases the bioavailability of EGFR ligands via stimulation of the cell-surface metalloproteinase ADAM17. Oncogene. 30:611–618. 2011. View Article : Google Scholar
12	Zheng X, Jiang F, Katakowski M, Lu Y and Chopp M: ADAM17 promotes glioma cell malignant phenotype. Mol Carcinog. 51:150–164. 2012. View Article : Google Scholar
13	Xiao LJ, Lin P, Lin F, Liu X, Qin W, Zou HF, Guo L, Liu W, Wang SJ and Yu XG: ADAM17 targets MMP-2 and MMP-9 via EGFR-MEK-ERK pathway activation to promote prostate cancer cell invasion. Int J Oncol. 40:1714–1724. 2012.
14	Zheng X, Jiang F, Katakowski M, Zhang ZG, Lu QE and Chopp M: ADAM17 promotes breast cancer cell malignant phenotype through EGFR-PI3K-AKT activation. Cancer Biol Ther. 8:1045–1054. 2009. View Article : Google Scholar : PubMed/NCBI
15	McGowan PM, McKiernan E, Bolster F, Ryan BM, Hill AD, McDermott EW, Evoy D, O'Higgins N, Crown J and Duffy MJ: ADAM-17 predicts adverse outcome in patients with breast cancer. Ann Oncol. 19:1075–1081. 2008. View Article : Google Scholar : PubMed/NCBI
16	Han X, Yang XF, Zhang XP, Sun Y, Zhao JH and Zhao GM: Expression of ADAM17 in tissue of breast cancer and its clinical significance. Modern Journal of Integrated Traditional Chinese and Western Medicine. 19:2486–2488. 2010.
17	Yang XF, Zhang XP, Zhao GM, Sun Y and Han X: Clinical significance of ADAM-17 protein expression in invasive breast cancer. Shandong Med J. 51:84–85. 2011.
18	Yang WJ, Zhang XP, Jiao JM, Zhao GM, Lu YQ and Hu BS: Inhibitory effects of ADAM17-siRNA on the invasion of human MCF-7 breast cancer. Tianjin Med J. 39:1045–1407. 2011.
19	Lu YQ, Zhao GM and Zhang XP: Effects of ADAM17-siRNA on the proliferation of human MCF-7 breast cancer. Modern J Integrated Traditional Chinese and Western Med. 20:3003–3005. 2011.
20	Brand TM, Iida M, Luthar N, Starr MM, Huppert EJ and Wheeler DL: Nuclear EGFR as a molecular target in cancer. Radiother Oncol. 108:370–377. 2013. View Article : Google Scholar
21	Dienstmann R, Braña I, Rodon J and Tabernero J: Toxicity as a biomarker of efficacy of molecular targeted therapies: Focus on EGFR and VEGF inhibiting anticancer drugs. Oncologist. 16:1729–1740. 2011. View Article : Google Scholar : PubMed/NCBI
22	Azuaje F, Tiemann K and Niclou SP: Therapeutic control and resistance of the EGFR-driven signaling network in glioblastoma. Cell Commun Signal. 13:232015. View Article : Google Scholar : PubMed/NCBI
23	Lee CC, Shiao HY, Wang WC and Hsieh HP: Small-molecule EGFR tyrosine kinase inhibitors for the treatment of cancer. Expert Opin Investig Drugs. 23:1333–1348. 2014. View Article : Google Scholar : PubMed/NCBI
24	Lorusso V, Forcignano R, Cinieri S, Tinelli A, Porcelli L, Quatrale AE and Chiuri VE: Which role for EGFR therapy in breast cancer? Front Biosci (Schol Ed). 4:31–42. 2012. View Article : Google Scholar
25	Lluch A, Eroles P and Perez-Fidalgo JA: Emerging EGFR antagonists for breast cancer. Expert Opin Emerg Drugs. 19:165–181. 2014. View Article : Google Scholar : PubMed/NCBI
26	Howe LR and Brown PH: Targeting the HER/EGFR/ErbB family to prevent breast cancer. Cancer Prev Res (Phila). 4:1149–1157. 2011. View Article : Google Scholar
27	Davis NM, Sokolosky M, Stadelman K, Abrams SL, Libra M, Candido S, Nicoletti F, Polesel J, Maestro R, D'Assoro A, et al: Deregulation of the EGFR/PI3K/PTEN/Akt/mTORC1 pathway in breast cancer: Possibilities for therapeutic intervention. Oncotarget. 5:4603–4650. 2014. View Article : Google Scholar : PubMed/NCBI
28	Li P, Torossian A, Zhang Q, Xu WC and Fu S: Inhibition of phosphoinositide 3-kinase enhances the cytotoxicity of AG1478, an epidermal growth factor receptor inhibitor, in breast cancer cells. Med Oncol. 29:3258–3264. 2012. View Article : Google Scholar : PubMed/NCBI
29	Caja L, Sancho P, Bertran E, Ortiz C, Campbell JS, Fausto N and Fabregat I: The tyrphostin AG1478 inhibits proliferation and induces death of liver tumor cells through EGF receptor-dependent and independent mechanisms. Biochem Pharmacol. 82:1583–1592. 2011. View Article : Google Scholar
30	Zhang YG, Du Q, Fang WG, Jin ML and Tian XX: Tyrphostin AG1478 suppresses proliferation and invasion of human breast cancer cells. Int J Oncol. 33:595–602. 2008.PubMed/NCBI
31	Ke XY, Wang Y, Xie ZQ, Liu ZQ, Zhang CF, Zhao Q and Yang DL: LY294002 enhances inhibitory effect of gemcitabine on proliferation of human pancreatic carcinoma PANC-1 cells. J Huazhong Univ Sci Technolog Med Sci. 33:57–62. 2013. View Article : Google Scholar : PubMed/NCBI
32	Tian S, Chang W, Du H, Bai J, Sun Z, Zhang Q, Wang H, Zhu G, Tao K and Long Y: The interplay between GRP78 expression and Akt activation in human colon cancer cells under celecoxib treatment. Anticancer Drugs. 26:964–973. 2015. View Article : Google Scholar : PubMed/NCBI
33	Zhang Y, Zheng L, Ding Y, Li Q, Wang R, Liu T, Sun Q, Yang H, Peng S, Wang W, et al: MiR-20a induces cell radioresistance by activating the PTEN/PI3K/Akt signaling pathway in hepatocellular carcinoma. Int J Radiat Oncol Biol Phys. 92:1132–1140. 2015. View Article : Google Scholar : PubMed/NCBI
34	Chen P, Wu J, Yuan Q, Jiang X and Huang H: The synergistic killing of AML cells co-cultured with HS-5 bone marrow stromal cells by As2O3 and the PI3K/Akt signaling pathway inhibitor LY294002. Pharmazie. 70:322–327. 2015.PubMed/NCBI
35	Wang Y and Lazo JS: Metastasis-associated phosphatase PRL-2 regulates tumor cell migration and invasion. Oncogene. 31:818–827. 2012. View Article : Google Scholar
36	Whitsett TG, Cheng E, Inge L, Asrani K, Jameson NM, Hostetter G, Weiss GJ, Kingsley CB, Loftus JC, Bremner R, et al: Elevated expression of Fn14 in non-small cell lung cancer correlates with activated EGFR and promotes tumor cell migration and invasion. Am J Pathol. 181:111–120. 2012. View Article : Google Scholar : PubMed/NCBI
37	Han J, Xie Y, Lan F, Yu Y, Liu W, Chen J, Zheng F, Ouyang X, Lin X, Lin Y, et al: Additive effects of EGF and IL-1β regulate tumor cell migration and invasion in gastric adenocarcinoma via activation of ERK1/2. Int J Oncol. 45:291–301. 2014.PubMed/NCBI
38	Kenny PA and Bissell MJ: Targeting TACE-dependent EGFR ligand shedding in breast cancer. J Clin Invest. 117:337–345. 2007. View Article : Google Scholar : PubMed/NCBI
39	Tanei T, Shimomura A, Shimazu K, Nakayama T, Kim SJ, Iwamoto T, Tamaki Y and Noguchi S: Prognostic significance of Ki67 index after neoadjuvant chemotherapy in breast cancer. Eur J Surg Oncol. 37:155–161. 2011. View Article : Google Scholar
40	Yoshioka T, Hosoda M, Yamamoto M, Taguchi K, Hatanaka KC, Takakuwa E, Hatanaka Y, Matsuno Y and Yamashita H: Prognostic significance of pathologic complete response and Ki67 expression after neoadjuvant chemotherapy in breast cancer. Breast Cancer. 22:185–191. 2015. View Article : Google Scholar