Lentiviral-mediated gene silencing of Notch-4 inhibits in vitro proliferation and perineural invasion of ACC-M cells

Chen,Wei; Zhang,Huali; Wang,Jiandong; Cao,Gang; Dong,Zhen; Su,Han; Zhou,Xiaojun; Zhang,Senlin

doi:10.3892/or.2013.2317

Lentiviral-mediated gene silencing of Notch-4 inhibits in vitro proliferation and perineural invasion of ACC-M cells

Authors:
- Wei Chen
- Huali Zhang
- Jiandong Wang
- Gang Cao
- Zhen Dong
- Han Su
- Xiaojun Zhou
- Senlin Zhang
View Affiliations

Affiliations: Department of Stomatology, Nanjing Jinling Hospital, Nanjing University, School of Medicine, Nanjing, Jiangsu, P.R. China, Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, Shaanxi, P.R. China, Department of Pathology, Nanjing Jinling Hospital, Nanjing University, School of Medicine, Nanjing, Jiangsu, P.R. China
Published online on: February 28, 2013 https://doi.org/10.3892/or.2013.2317
Pages: 1797-1804

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

Salivary adenoid cystic carcinoma (SACC) is a common type of salivary gland cancer. The poor long-term prognosis for patients with SACC is mainly due to local recurrence, perineural invasion (PNI) and distant metastasis. Notch signaling plays a critical role in determining cell fate such as proliferation, differentiation and apoptosis. Accumulating evidence indicates that aberrant Notch-4 expression has a tumor-promoting function in SACC. In the present study, we used lentiviral-mediated RNA interference (RNAi) targeted against Notch-4 to determine the effects of decreased levels of this protein in the human highly metastatic adenoid cystic carcinoma cell line ACC-M. Furthermore, the proliferative capability as well as the PNI potential of the treated cells were observed in vitro. Our studies demonstrated that RNAi directed against Notch-4 markedly decreased Notch-4 gene expression, resulting in the inhibition of cell proliferation, and G0/G1 to S phase arrest in ACC-M cells. Knockdown of Notch-4 also resulted in a decrease in the in vitro PNI activity in ACC-M cells. To conclude, RNAi targeting against Notch-4 induces the suppression of cell growth and inhibition of PNI in vitro in ACC-M cells. Notch-4 may play an important role in regulating proliferation and PNI activity of SACC.

View Figures

View References

1	Renehan A, Gleave EN, Hancock BD, Smith P and McGurk M: Long-term follow-up of over 1000 patients with salivary gland tumours treated in a single centre. Br J Surg. 83:1750–1754. 1996. View Article : Google Scholar : PubMed/NCBI
2	Anderson JN Jr, Beenken SW, Crowe R, Soong SJ, Peters G, Maddox WA and Urist MM: Prognostic factors in minor salivary gland cancer. Head Neck. 17:480–486. 1995. View Article : Google Scholar : PubMed/NCBI
3	Van der Wal JE, Becking AG, Snow GB and van der Waal I: Distant metastases of adenoid cystic carcinoma of the salivary glands and the value of diagnostic examinations during follow-up. Head Neck. 24:779–783. 2002.PubMed/NCBI
4	Ramer N, Wu H, Sabo E, Ramer Y, Emanuel P, Orta L and Burstein DE: Prognostic value of quantitative p63 immuno- staining in adenoid cystic carcinoma of salivary gland assessed by computerized image analysis. Cancer. 116:77–83. 2010.PubMed/NCBI
5	Van der Wal JE, Snow GB and van der Waal I: Intraoral adenoid cystic carcinoma. The presence of perineural spread in relation to site, size, local extension, and metastatic spread in 22 cases. Cancer. 66:2031–2033. 1990.PubMed/NCBI
6	Vrielinck L, Ostyn F, van Damme B, van den Boqaert W and Fossion E: The significance of perineural spread in adenoid cystic carcinoma of the major and minor salivary glands. Int J Oral Maxillofac Surg. 17:190–193. 1988. View Article : Google Scholar : PubMed/NCBI
7	Newlin HE, Morris CG, Amdur RJ and Mendenhall WM: Neurotropic melanoma of the head and neck with clinical perineural invasion. Am J Clin Oncol. 28:399–402. 2005. View Article : Google Scholar : PubMed/NCBI
8	Ayala GE, Dai H, Ittmann M, Li R, Powell M, Frolov A, Wheeler TM, Thompson TC and Rowley D: Growth and survival mechanisms associated with perineural invasion in prostate cancer. Cancer Res. 64:6082–6090. 2004. View Article : Google Scholar : PubMed/NCBI
9	Pour PM, Bell RH and Batra SK: Neural invasion in the staging of pancreatic cancer. Pancreas. 26:322–325. 2003. View Article : Google Scholar : PubMed/NCBI
10	Faqan JJ, Collins B, Barnes L, D'Amico F, Myers EN and Johnson JT: Perineural invasion in squamous cell carcinoma of the head and neck. Arch Otolaryngol Head Neck Surg. 124:637–640. 1998. View Article : Google Scholar : PubMed/NCBI
11	Rahima B, Shingaki S, Nagata M and Saito C: Prognostic significance of perineural invasion in oral and oropharyngeal carcinoma. Oral Surg Oral Med Oral Pathol Radiol Endod. 97:423–431. 2004. View Article : Google Scholar : PubMed/NCBI
12	Go MJ, Eastman DS and Artavanis-Tsakonas S: Cell proliferation control by Notch signaling in Drosophila development. Development. 125:2031–2040. 1998.PubMed/NCBI
13	Shelly LL, Fuchs C and Miele L: Notch-1 inhibits apoptosis in murine erythroleukemia cells and is necessary for differentiation induced by hybrid polar compounds. J Cell Biochem. 73:164–175. 1999. View Article : Google Scholar : PubMed/NCBI
14	Bigas A, Martin DI and Milner LA: Notch1 and Notch2 inhibit myeloid differentiation in response to different cytokines. Mol Cell Biol. 18:2324–2333. 1998.PubMed/NCBI
15	Mumm JS and Kopan R: Notch signaling: from the outside in. Dev Biol. 228:151–165. 2000. View Article : Google Scholar : PubMed/NCBI
16	Stockhausen MT, Sjölund J and Axelson H: Regulation of the Notch target gene Hes-1 by TGFalpha induced Ras/MAPK signaling in human neuroblastoma cells. Exp Cell Res. 310:218–228. 2005. View Article : Google Scholar : PubMed/NCBI
17	Yoon JH, Kim SA, Kwon SM, Park JH, Park HS, Kim YC, Yoon JH and Ahn SG: 5′-Nitro-indirubinoxime induces G1 cell cycle arrest and apoptosis in salivary gland adenocarcinoma cells through the inhibition of Notch-1 signaling. Biochim Biophys Acta. 1800:352–358. 2010.
18	Yao J, Duan L, Fan M, Yuan J and Wu X: Notch1 induces cell cycle arrest and apoptosis in human cervical cancer cells: involvement of nuclear factor kappa B inhibition. Int J Gynecol Cancer. 17:502–510. 2007. View Article : Google Scholar : PubMed/NCBI
19	Duan L, Yao J, Wu X and Fan M: Growth suppression induced by Notch1 activation involves Wnt-β-catenin down-regulation in human tongue carcinoma cells. Biol Cell. 98:479–490. 2006.PubMed/NCBI
20	Leethanakul C, Patel V, Gillespie J, Pallente M, Ensley JF, Koontongkaew S, Liotta LA, Emmert-Buck M and Gutkind JS: Distinct pattern of expression of differentiation and growth-related genes in squamous cell carcinomas of the head and neck revealed by the use of laser capture microdissection and cDNA arrays. Oncogene. 19:3220–3224. 2000. View Article : Google Scholar : PubMed/NCBI
21	Nefedova Y, Sullivan DM, Bolick SC, Dalton WS and Gabrilovich DI: Inhibition of Notch signaling induces apoptosis of myeloma cells and enhances sensitivity to chemotherapy. Blood. 111:2220–2229. 2008. View Article : Google Scholar : PubMed/NCBI
22	Zang S, Chen F, Dai J, Guo D, Tse W, Qu X, Ma D and Ji C: RNAi-mediated knockdown of Notch-1 leads to cell growth inhibition and enhanced chemosensitivity in human breast cancer. Oncol Rep. 23:893–899. 2010.PubMed/NCBI
23	Gu F, Ma Y, Zhang Z, Zhao J, Kobayashi H, Zhang L and Fu L: Expression of Stat3 and Notch1 is associated with cisplatin resistance in head and neck squamous cell carcinoma. Oncol Rep. 23:671–676. 2010.PubMed/NCBI
24	Ding LC, She L, Zheng DL, Huang QL, Wang JF, Zheng FF and Lu YG: Notch-4 contributes to the metastasis of salivary adenoid cystic carcinoma. Oncol Rep. 24:363–368. 2010.PubMed/NCBI
25	Chen W, Zhang HL, Shao XJ, Jiang YG, Zhao XG, Gao X, Li JH, Yang J, Zhang YF, Liu BL and Sun MY: Gene expression profile of salivary adenoid cystic carcinoma associated with perineural invasion. Tohoku J Exp Med. 212:319–334. 2007. View Article : Google Scholar : PubMed/NCBI
26	Guan XF, Qiu WL, He RG and Zhou XJ: Selection of adenoid cystic carcinoma cell clone highly metastatic to the lung: an experimental study. Int J Oral Maxillofac Surg. 26:116–119. 1997. View Article : Google Scholar : PubMed/NCBI
27	Tuschl T: Expanding small RNA interference. Nat Biotechnol. 20:446–448. 2002. View Article : Google Scholar : PubMed/NCBI
28	Reynolds A, Leake D, Boese Q, Scaringe S, Marshall WS and Khvorova A: Rational siRNA design for RNA interference. Nat Biotechnol. 22:326–330. 2004. View Article : Google Scholar : PubMed/NCBI
29	Wang L, Sun M, Jiang Y, Yang L, Lei D, Lu C, Zhao Y, Zhang P, Yang Y and Li J: Nerve growth factor and tyrosine kinase A in human salivary adenoid cystic carcinoma: expression patterns and effects on in vitro invasive behavior. J Oral Maxillofac Surg. 64:636–641. 2006. View Article : Google Scholar
30	Chen W, Zhang HL, Jiang YG, Li JH, Liu BL and Sun MY: Inhibition of CD146 gene expression via RNA interference reduces in vitro perineural invasion on ACC-M cell. J Oral Pathol Med. 38:198–205. 2009. View Article : Google Scholar : PubMed/NCBI
31	Mohr OL: Character changes caused by mutation of an entire region of a chromosome in Drosophila. Genetics. 4:275–282. 1919.PubMed/NCBI
32	Miele L and Osborne B: Arbiter of differentiation and death: Notch signaling meets apoptosis. J Cell Physiol. 181:393–409. 1999. View Article : Google Scholar : PubMed/NCBI
33	Cheng P and Gabrilovich D: Notch signaling in differentiation and function of dendritic cells. Immunol Res. 41:1–14. 2008. View Article : Google Scholar : PubMed/NCBI
34	Baker NE: Notch signaling in the nervous system. Pieces still missing from the puzzle. Bioessays. 22:264–273. 2000. View Article : Google Scholar : PubMed/NCBI
35	Artavanis-Tsakonas S, Rand MD and Lake RJ: Notch signaling: cell fate control and signal integration in development. Science. 284:770–776. 1999. View Article : Google Scholar : PubMed/NCBI
36	Milner LA and Bigas A: Notch as a mediator of cell fate determination in hematopoiesis: evidence and speculation. Blood. 93:2431–2448. 1999.PubMed/NCBI
37	Kopan R and Ilagan MX: The canonical Notch signaling pathway: unfolding the activation mechanism. Cell. 137:216–233. 2009. View Article : Google Scholar : PubMed/NCBI
38	Clementz AG, Rogowski A, Pandya K, Miele L and Osipo C: Notch-1 and Notch-4 are novel gene targets of PEA3 in breast cancer: novel therapeutic implications. Breast Cancer Res. 13:R632011. View Article : Google Scholar : PubMed/NCBI
39	Zamore PD, Tuschl T, Sharp PA and Bartel DP: RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell. 101:25–33. 2000. View Article : Google Scholar : PubMed/NCBI
40	Yamamoto T, Miyoshi H, Yamamoto N, Yamamoto N, Inoue J and Tsunetsugu-Yokota Y: Lentivirus vectors expressing short hairpin RNAs against the U3-overlapping region of HIV nef inhibit HIV replication and infectivity in primary macrophages. Blood. 108:3305–3312. 2006. View Article : Google Scholar : PubMed/NCBI
41	Desclaux M, Teigell M, Amar L, Vogel R, Gimenez Y, Ribotta M, Privat A and Mallet J: A novel and efficient gene transfer strategy reduces glial reactivity and improves neuronal survival and axonal growth in vitro. PLoS One. 4:e62272009. View Article : Google Scholar : PubMed/NCBI