VEGF-D-enhanced lymph node metastasis of ovarian cancer is reversed by vesicular stomatitis virus matrix protein

Qi,Xiaorong; Du,Licheng; Chen,Xiancheng; Chen,Lijuan; Yi,Tao; Chen,Xiang; Wen,Yanjun; Wei,Yuquan; Zhao,Xia

doi:10.3892/ijo.2016.3527

VEGF-D-enhanced lymph node metastasis of ovarian cancer is reversed by vesicular stomatitis virus matrix protein

Authors:
- Xiaorong Qi
- Licheng Du
- Xiancheng Chen
- Lijuan Chen
- Tao Yi
- Xiang Chen
- Yanjun Wen
- Yuquan Wei
- Xia Zhao
View Affiliations

Affiliations: Department of Gynecology and Obstetrics, Key Laboratory of Obstetric and Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second Hospital, State Key Laboratory of Biotherapy, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Provincial Hospital Affiliated to Shandong University, Jinan, P.R. China, State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: May 17, 2016 https://doi.org/10.3892/ijo.2016.3527
Pages: 123-132
Copyright: © Qi 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

Lymphatic metastasis is a poor prognostic factor in ovarian cancer, which correlates to the majority of cancer deaths. Matrix protein (MP) of vesicular stomatitis virus (VSV) exhibits potent antitumor and antiangiogenic activities through inducing apoptosis and inhibiting angiogenesis. In this study, the antitumor and antimetastatic effects of MP were further investigated. Wild-type SKOV3 (WT-SK) cells were successfully transfected with empty vector pcDNA3.1 plasmid, or pcDNA3.1-VEGF-D recombinant plasmid to construct cell lines named EV-SK, and VEGFD-SK, respectively. Inhibition of VEGFD-SK cell migration and invasion was detected by Transwell and wound healing assay. Then, lymphogenous metastatic model of ovarian cancer was established by injecting VEGFD-SK cells subcutaneously into the left hindlimb claw pad of nude mice. The inducted apoptotic effect of MP on VEGFD-SK cells were assessed by flow analysis and Hoechst-33258 staining, respectively, in vitro. The in vivo antitumor and antiangiogenic activities of MP gene were evaluated with lymphogenous metastatic model of ovarian cancer. Tumor volume and lymphatic metastasis rates were measured. Lymphatic vessels were delineated using Evan's blue and LYVE-1 staining. Expression of VEGF-D and MMP-2 were evaluated by immunostaining. Apoptosis of tumor cells was analyzed by Hoechst-33258 staining. Mice bearing VEGFD-SK tumor cells displayed more rapid tumorigenesis, higher lymphogenous metastatic tendency and increased lymphatic vessel density compared with the mice bearing WT-SK or EV-SK cells. However, VEGF-D-enhanced metastasis was evidently reversed by MP. MP significantly reduced the invasion of VEGFD-SK cells, tumor volume, lymphatic metastasis rates and lymphatic vessel density compared with control groups (P<0.05), accompanied with down-expression of VEGF-D and MMP-2 and increased apoptosis. Our data indicate that MP has strong antitumor and antimetastatic abilities, and it may be a promising therapeutic strategy against the lymphatic metastasis of human ovarian cancer.

View Figures

View References

1	Edwards BK, Brown ML, Wingo PA, Howe HL, Ward E, Ries LA, Schrag D, Jamison PM, Jemal A, Wu XC, et al: Annual report to the nation on the status of cancer, 1975–2002 featuring population-based trends in cancer treatment. J Natl Cancer Inst. 97:1407–1427. 2005. View Article : Google Scholar : PubMed/NCBI
2	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
3	Concin N, Hefler L, van Bavel J, Mueller-Holzner E, Zeimet A, Daxenbichler G, Speiser P, Hacker N and Marth C: Biological markers in pT1 and pT2 ovarian cancer with lymph node metastases. Gynecol Oncol. 89:9–15. 2003. View Article : Google Scholar : PubMed/NCBI
4	Rao G, Crispens M and Rothenberg ML: Intraperitoneal chemotherapy for ovarian cancer: Overview and perspective. J Clin Oncol. 25:2867–2872. 2007. View Article : Google Scholar : PubMed/NCBI
5	Ayhan A, Gultekin M, Taskiran C, Celik NY, Usubutun A, Kucukali T and Yuce K: Lymphatic metastasis in epithelial ovarian carcinoma with respect to clinicopathological variables. Gynecol Oncol. 97:400–404. 2005. View Article : Google Scholar : PubMed/NCBI
6	di Re F, Baiocchi G, Fontanelli R, Grosso G, Cobellis L, Raspagliesi F and di Re E: Systematic pelvic and paraaortic lymphadenectomy for advanced ovarian cancer: Prognostic significance of node metastases. Gynecol Oncol. 62:360–365. 1996. View Article : Google Scholar : PubMed/NCBI
7	Millauer B, Wizigmann-Voos S, Schnürch H, Martinez R, Møller NP, Risau W and Ullrich A: High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell. 72:835–846. 1993. View Article : Google Scholar : PubMed/NCBI
8	Kaipainen A, Korhonen J, Mustonen T, van Hinsbergh VW, Fang GH, Dumont D, Breitman M and Alitalo K: Expression of the fms-like tyrosine kinase 4 gene becomes restricted to lymphatic endothelium during development. Proc Natl Acad Sci USA. 92:3566–3570. 1995. View Article : Google Scholar : PubMed/NCBI
9	Stacker SA, Caesar C, Baldwin ME, Thornton GE, Williams RA, Prevo R, Jackson DG, Nishikawa S, Kubo H and Achen MG: VEGF-D promotes the metastatic spread of tumor cells via the lymphatics. Nat Med. 7:186–191. 2001. View Article : Google Scholar : PubMed/NCBI
10	Wang J, Guo Y, Wang B, Bi J, Li K, Liang X, Chu H and Jiang H: Lymphatic microvessel density and vascular endothelial growth factor-C and -D as prognostic factors in breast cancer: A systematic review and meta-analysis of the literature. Mol Biol Rep. 39:11153–11165. 2012. View Article : Google Scholar : PubMed/NCBI
11	Black BL and Lyles DS: Vesicular stomatitis virus matrix protein inhibits host cell-directed transcription of target genes in vivo. J Virol. 66:4058–4064. 1992.PubMed/NCBI
12	Gerlier D and Lyles DS: Interplay between innate immunity and negative-strand RNA viruses: Towards a rational model. Microbiol Mol Biol Rev. 75:468–490. 2011. View Article : Google Scholar : PubMed/NCBI
13	Ahmed M and Lyles DS: Effect of vesicular stomatitis virus matrix protein on transcription directed by host RNA polymerases I, II, and III. J Virol. 72:8413–8419. 1998.PubMed/NCBI
14	Yuan H, Puckett S and Lyles DS: Inhibition of host transcription by vesicular stomatitis virus involves a novel mechanism that is independent of phosphorylation of TATA-binding protein (TBP) or association of TBP with TBP-associated factor subunits. J Virol. 75:4453–4458. 2001. View Article : Google Scholar : PubMed/NCBI
15	Ahmed M, McKenzie MO, Puckett S, Hojnacki M, Poliquin L and Lyles DS: Ability of the matrix protein of vesicular stomatitis virus to suppress beta interferon gene expression is genetically correlated with the inhibition of host RNA and protein synthesis. J Virol. 77:4646–4657. 2003. View Article : Google Scholar : PubMed/NCBI
16	Petersen JM, Her LS, Varvel V, Lund E and Dahlberg JE: The matrix protein of vesicular stomatitis virus inhibits nucleocytoplasmic transport when it is in the nucleus and associated with nuclear pore complexes. Mol Cell Biol. 20:8590–8601. 2000. View Article : Google Scholar : PubMed/NCBI
17	Dunn EF and Connor JH: Dominant inhibition of Akt/protein kinase B signaling by the matrix protein of a negative-strand RNA virus. J Virol. 85:422–431. 2011. View Article : Google Scholar :
18	Zhong Q, Wen YJ, Yang HS, Luo H, Fu AF, Yang F, Chen LJ, Chen X, Qi XR, Lin HG, et al: Efficient inhibition of cisplatin-resistant human ovarian cancer growth and prolonged survival by gene transferred vesicular stomatitis virus matrix protein in nude mice. Ann Oncol. 19:1584–1591. 2008. View Article : Google Scholar : PubMed/NCBI
19	Gou M, Men K, Zhang J, Li Y, Song J, Luo S, Shi H, Wen Y, Guo G, Huang M, et al: Efficient inhibition of C-26 colon carcinoma by VSVMP gene delivered by biodegradable cationic nanogel derived from polyethyleneimine. ACS Nano. 4:5573–5584. 2010. View Article : Google Scholar : PubMed/NCBI
20	Ahmed M, Puckett S and Lyles DS: Susceptibility of breast cancer cells to an oncolytic matrix (M) protein mutant of vesicular stomatitis virus. Cancer Gene Ther. 17:883–892. 2010. View Article : Google Scholar : PubMed/NCBI
21	Zhou Y, Wen F, Zhang P, Tang R and Li Q: Matrix protein of vesicular stomatitis virus: A potent inhibitor of vascular endothelial growth factor and malignant ascites formation. Cancer Gene Ther. 20:178–185. 2013. View Article : Google Scholar : PubMed/NCBI
22	Wen J, Fu AF, Chen LJ, Xie XJ, Yang GL, Chen XC, Wang YS, Li J, Chen P, Tang MH, et al: Liposomal honokiol inhibits VEGF-D-induced lymphangiogenesis and metastasis in xenograft tumor model. Int J Cancer. 124:2709–2718. 2009. View Article : Google Scholar : PubMed/NCBI
23	Lin X, Chen X, Wei Y, Zhao J, Fan L, Wen Y, Wu H and Zhao X: Efficient inhibition of intraperitoneal human ovarian cancer growth and prolonged survival by gene transfer of vesicular stomatitis virus matrix protein in nude mice. Gynecol Oncol. 104:540–546. 2007. View Article : Google Scholar
24	Chen X, Wang X, Wang Y, Yang L, Hu J, Xiao W, Fu A, Cai L, Li X and Ye X: Improved tumor-targeting drug delivery and therapeutic efficacy by cationic liposome modified with truncated bFGF peptide. J Control Release. 145:17–25. 2010. View Article : Google Scholar : PubMed/NCBI
25	Zhou L, Du L, Chen X, Li X, Li Z, Wen Y, Li Z, He X, Wei Y, Zhao X, et al: The antitumor and antimetastatic effects of N-trimethyl chitosan-encapsulated camptothecin on ovarian cancer with minimal side effects. Oncol Rep. 24:941–948. 2010.PubMed/NCBI
26	Jussila L and Alitalo K: Vascular growth factors and lymphangiogenesis. Physiol Rev. 82:673–700. 2002. View Article : Google Scholar : PubMed/NCBI
27	Kabashima A, Maehara Y, Kakeji Y, Baba H, Koga T and Sugimachi K: Clinicopathological features and overexpression of matrix metalloproteinases in intramucosal gastric carcinoma with lymph node metastasis. Clin Cancer Res. 6:3581–3584. 2000.PubMed/NCBI
28	Mitra A, Chakrabarti J, Banerji A and Chatterjee A: Cell membrane-associated MT1-MMP dependent activation of MMP-2 in SiHa (human cervical cancer) cells. J Environ Pathol Toxicol Oncol. 25:655–666. 2006. View Article : Google Scholar
29	Griffioen AW: Lymphangiogenesis factors: a target for therapy? Blood. 113:4135–4136. 2009. View Article : Google Scholar : PubMed/NCBI
30	Kubo H, Fujiwara T, Jussila L, Hashi H, Ogawa M, Shimizu K, Awane M, Sakai Y, Takabayashi A, Alitalo K, et al: Involvement of vascular endothelial growth factor receptor-3 in maintenance of integrity of endothelial cell lining during tumor angiogenesis. Blood. 96:546–553. 2000.PubMed/NCBI
31	Pytowski B, Goldman J, Persaud K, Wu Y, Witte L, Hicklin DJ, Skobe M, Boardman KC and Swartz MA: Complete and specific inhibition of adult lymphatic regeneration by a novel VEGFR-3 neutralizing antibody. J Natl Cancer Inst. 97:14–21. 2005. View Article : Google Scholar : PubMed/NCBI
32	Majumder M, Tutunea-Fatan E, Xin X, Rodriguez-Torres M, Torres-Garcia J, Wiebe R, Timoshenko AV, Bhattacharjee RN, Chambers AF and Lala PK: Co-expression of α9β1 integrin and VEGF-D confers lymphatic metastatic ability to a human breast cancer cell line MDA-MB-468LN. PLoS One. 7:e350942012. View Article : Google Scholar
33	Renyi-Vamos F, Tovari J, Fillinger J, Timar J, Paku S, Kenessey I, Ostoros G, Agocs L, Soltesz I and Dome B: Lymphangiogenesis correlates with lymph node metastasis, prognosis, and angiogenic phenotype in human non-small cell lung cancer. Clin Cancer Res. 11:7344–7353. 2005. View Article : Google Scholar : PubMed/NCBI
34	Hall FT, Freeman JL, Asa SL, Jackson DG and Beasley NJ: Intratumoral lymphatics and lymph node metastases in papillary thyroid carcinoma. Arch Otolaryngol Head Neck Surg. 129:716–719. 2003. View Article : Google Scholar : PubMed/NCBI
35	Dadras SS, Paul T, Bertoncini J, Brown LF, Muzikansky A, Jackson DG, Ellwanger U, Garbe C, Mihm MC and Detmar M: Tumor lymphangiogenesis: A novel prognostic indicator for cutaneous melanoma metastasis and survival. Am J Pathol. 162:1951–1960. 2003. View Article : Google Scholar : PubMed/NCBI
36	Bono P, Wasenius VM, Heikkilä P, Lundin J, Jackson DG and Joensuu H: High LYVE-1-positive lymphatic vessel numbers are associated with poor outcome in breast cancer. Clin Cancer Res. 10:7144–7149. 2004. View Article : Google Scholar : PubMed/NCBI
37	O'Grady A, Dunne C, O'Kelly P, Murphy GM, Leader M and Kay E: Differential expression of matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in non-melanoma skin cancer: Implications for tumour progression. Histopathology. 51:793–804. 2007. View Article : Google Scholar : PubMed/NCBI
38	Taniwaki K, Fukamachi H, Komori K, Ohtake Y, Nonaka T, Sakamoto T, Shiomi T, Okada Y, Itoh T, Itohara S, et al: Stroma-derived matrix metalloproteinase (MMP)-2 promotes membrane type 1-MMP-dependent tumor growth in mice. Cancer Res. 67:4311–4319. 2007. View Article : Google Scholar : PubMed/NCBI
39	Egeblad M and Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2:161–174. 2002. View Article : Google Scholar : PubMed/NCBI
40	Nathanson SD, Anaya P, Avery M, Hetzel FW, Sarantou T and Havstad S: Sentinel lymph node metastasis in experimental melanoma: Relationships among primary tumor size, lymphatic vessel diameter and 99mTc-labeled human serum albumin clearance. Ann Surg Oncol. 4:161–168. 1997. View Article : Google Scholar : PubMed/NCBI
41	Carter CL, Allen C and Henson DE: Relation of tumor size, lymph node status, and survival in 24,740 breast cancer cases. Cancer. 63:181–187. 1989. View Article : Google Scholar : PubMed/NCBI
42	Nagamoto N, Saito Y, Ohta S, Sato M, Kanma K, Sagawa M, Takahashi S, Usuda K, Nakada T and Hashimoto K: Relationship of lymph node metastasis to primary tumor size and microscopic appearance of roentgenographically occult lung cancer. Am J Surg Pathol. 13:1009–1013. 1989. View Article : Google Scholar : PubMed/NCBI