Targeted CDX2 expression inhibits aggressive phenotypes of colon cancer cells in vitro and in vivo

Zheng,Jianbao; He,Sai; Qi,Jie; Wang,Xiaolong; Yu,Junhui; Wu,Yunhua; Gao,Qi; Wang,Kai; Sun,Xuejun

doi:10.3892/ijo.2017.4040

Targeted CDX2 expression inhibits aggressive phenotypes of colon cancer cells in vitro and in vivo

Authors:
- Jianbao Zheng
- Sai He
- Jie Qi
- Xiaolong Wang
- Junhui Yu
- Yunhua Wu
- Qi Gao
- Kai Wang
- Xuejun Sun
View Affiliations

Affiliations: Department of General Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Department of Breast Surgery, Shaanxi Provincial Tumor Hospital, Xi'an, Shaanxi 710061, P.R. China, Second Department of Cardiovascular Medicine, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710068, P.R. China, Department of Tumor Surgery, The Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China
Published online on: June 13, 2017 https://doi.org/10.3892/ijo.2017.4040
Pages: 478-488
Copyright: © Zheng 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

Loss of caudal type homeobox 2 (CDX2) is associated with the development of human colorectal cancer, while human telomerase reverse transcriptase (hTERT) frequently occurs in variety of human cancers. We investigated the effects of restoration of CDX2 expression using a hypoxia-inducible hTERT promoter-driven vector (pLVX-5HRE-hTERTp-CDX2-3FLAG) on colon cancer cell viability, cell cycle distribution, apoptosis, colony formation, invasion ability and xenograft tumor growth in nude mice. CDX2 overexpression significantly inhibited viability, colony formation, and the invasion and migration ability of LoVo cells, and induced cell cycle arrest and apoptosis in vitro, especially under hypoxic culture conditions. Overexpression of CDX2 under normoxic conditions significantly suppressed the expression of TGF-β, cyclin D1, uPA, MMP-9, MMP-2, and Bcl-2, and stimulated the expression of collagen IV, laminin-1, and Bax. Overexpression of CDX2 reduced colon cancer xenograft tumor formation in nude mice which was associated with downregulation of Ki-67. In conclusion, overexpression of CDX2 using a hypoxia-inducible hTERT promoter-driven vector suppressed malignant progression of colon cancer cells in vitro and in vivo. These results suggest that pLVX-5HRE-hTERTp-CDX2-3FLAG gene therapy may be a promising novel approach to treat colon cancer.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 66:7–30. 2016. View Article : Google Scholar : PubMed/NCBI
2	Shayakhmetov DM, Di Paolo NC and Mossman KL: Recognition of virus infection and innate host responses to viral gene therapy vectors. Mol Ther. 18:1422–1429. 2010. View Article : Google Scholar : PubMed/NCBI
3	Chen EQ, Song XQ, Wang YL, Zhou TY, Bai L, Liu L, Liu C, Cheng X and Tang H: Construction of a highly-active, liver-specific transcriptional regulatory element through combination of the albumin promoter and α-fetoprotein enhancer. Plasmid. 65:125–131. 2011. View Article : Google Scholar
4	Dong K, Wang R, Wang X, Lin F, Shen JJ, Gao P and Zhang HZ: Tumor-specific RNAi targeting eIF4E suppresses tumor growth, induces apoptosis and enhances cisplatin cytotoxicity in human breast carcinoma cells. Breast Cancer Res Treat. 113:443–456. 2009. View Article : Google Scholar
5	Akhavan-Niaki H and Samadani AA: Molecular insight in gastric cancer induction: An overview of cancer stemness genes. Cell Biochem Biophys. 68:463–473. 2014. View Article : Google Scholar
6	Natoli M, Christensen J, El-Gebali S, Felsani A and Anderle P: The role of CDX2 in Caco-2 cell differentiation. Eur J Pharm Biopharm. 85:20–25. 2013. View Article : Google Scholar : PubMed/NCBI
7	Lin ME, Huang D, Deng BH, Lv YS, Rong L and Yao YS: Expression and functional role of Cdx2 in intestinal metaplasia of cystitis glandularis. J Urol. 190:1083–1089. 2013. View Article : Google Scholar : PubMed/NCBI
8	Gross I, Duluc I, Benameur T, Calon A, Martin E, Brabletz T, Kedinger M, Domon-Dell C and Freund JN: The intestine-specific homeobox gene Cdx2 decreases mobility and antagonizes dissemination of colon cancer cells. Oncogene. 27:107–115. 2008. View Article : Google Scholar
9	Aoki K, Tamai Y, Horiike S, Oshima M and Taketo MM: Colonic polyposis caused by mTOR-mediated chromosomal instability in Apc⁺/Delta716 Cdx2⁺/⁻ compound mutant mice. Nat Genet. 35:323–330. 2003. View Article : Google Scholar : PubMed/NCBI
10	Chawengsaksophak K, James R, Hammond VE, Köntgen F and Beck F: Homeosis and intestinal tumours in Cdx2 mutant mice. Nature. 386:84–87. 1997. View Article : Google Scholar : PubMed/NCBI
11	Olsen AK, Coskun M, Bzorek M, Kristensen MH, Danielsen ET, Jørgensen S, Olsen J, Engel U, Holck S and Troelsen JT: Regulation of APC and AXIN2 expression by intestinal tumor suppressor CDX2 in colon cancer cells. Carcinogenesis. 34:1361–1369. 2013. View Article : Google Scholar : PubMed/NCBI
12	Hong KD, Lee D, Lee Y, Lee SI and Moon HY: Reduced CDX2 expression predicts poor overall survival in patients with colorectal cancer. Am Surg. 79:353–360. 2013.PubMed/NCBI
13	Zheng JB, Sun XJ, Qi J, Li SS, Wang W, Ren HL, Tian Y, Lu SY and Du JK: Effects of homeodomain protein CDX2 expression on the proliferation and migration of lovo colon cancer cells. Pathol Oncol Res. 17:743–751. 2011. View Article : Google Scholar : PubMed/NCBI
14	Wang W, Jin B, Li W, Xu CX, Cui FA, Liu B, Yan YF, Liu XX and Wang XL: Targeted antitumor effect induced by hTERT promoter mediated ODC antisense adenovirus. Mol Biol Rep. 37:3239–3247. 2010. View Article : Google Scholar
15	Bougel S, Renaud S, Braunschweig R, Loukinov D, Morse HC III, Bosman FT, Lobanenkov V and Benhattar J: PAX5 activates the transcription of the human telomerase reverse transcriptase gene in B cells. J Pathol. 220:87–96. 2010. View Article : Google Scholar
16	Zhang P, Tan J, Yang DB, Luo ZC, Luo S, Chen P, Sun P, Zhou Y, Chen XC, Wei YQ, et al: Gene therapy using the human telomerase catalytic subunit gene promoter enables targeting of the therapeutic effects of vesicular stomatitis virus matrix protein against human lung adenocarcinoma. Exp Ther Med. 4:859–864. 2012.PubMed/NCBI
17	Hioki M, Kagawa S and Fujiwara T, Sakai R, Kojima T, Watanabe Y, Hashimoto Y, Uno F, Tanaka N and Fujiwara T: Combination of oncolytic adenovirotherapy and Bax gene therapy in human cancer xenografted models. Potential merits and hurdles for combination therapy. Int J Cancer. 122:2628–2633. 2008. View Article : Google Scholar : PubMed/NCBI
18	Gout S and Huot J: Role of cancer microenvironment in metastasis: Focus on colon cancer. Cancer Microenviron. 1:69–83. 2008. View Article : Google Scholar
19	Jubb AM, Buffa FM and Harris AL: Assessment of tumour hypoxia for prediction of response to therapy and cancer prognosis. J Cell Mol Med. 14:18–29. 2010. View Article : Google Scholar
20	Law AY, Ching LY, Lai KP and Wong CK: Identification and characterization of the hypoxia-responsive element in human stanniocalcin-1 gene. Mol Cell Endocrinol. 314:118–127. 2010. View Article : Google Scholar
21	Zhang J, Shi Q, Chen X, Yang P, Qi C, Zhang J, Lu H, Liu J, Jiao Q, Zhao L, et al: Hypoxia-regulated neurotrophin-3 expression by multicopy hypoxia response elements reduces apoptosis in PC12 cells. Int J Mol Med. 30:1173–1179. 2012.PubMed/NCBI
22	Hu J, Stiehl DP, Setzer C, Wichmann D, Shinde DA, Rehrauer H, Hradecky P, Gassmann M and Gorr TA: Interaction of HIF and USF signaling pathways in human genes flanked by hypoxia-response elements and E-box palindromes. Mol Cancer Res. 9:1520–1536. 2011. View Article : Google Scholar : PubMed/NCBI
23	Zheng J, Sun X, Wang W and Lu S: Hypoxia-inducible factor-1α modulates the down-regulation of the homeodomain protein CDX2 in colorectal cancer. Oncol Rep. 24:97–104. 2010.PubMed/NCBI
24	Shibata T, Giaccia AJ and Brown JM: Development of a hypoxia-responsive vector for tumor-specific gene therapy. Gene Ther. 7:493–498. 2000. View Article : Google Scholar : PubMed/NCBI
25	Zhou PH, Zheng JB, Wei GB, Wang XL, Wang W, Chen NZ, Yu JH, Yao JF, Wang H, Lu SY, et al: Lentivirus-mediated RASSF1A expression suppresses aggressive phenotypes of gastric cancer cells in vitro and in vivo. Gene Ther. 22:793–801. 2015. View Article : Google Scholar : PubMed/NCBI
26	He S, Sun XJ, Zheng JB, Qi J, Chen NZ, Wang W, Wei GB, Liu D, Yu JH, Lu SY, et al: Recombinant lentivirus with enhanced expression of caudal-related homeobox protein 2 inhibits human colorectal cancer cell proliferation in vitro. Mol Med Rep. 12:1838–1844. 2015.PubMed/NCBI
27	Ji J and Zheng PS: Expression of Sox2 in human cervical carcinogenesis. Hum Pathol. 41:1438–1447. 2010. View Article : Google Scholar : PubMed/NCBI
28	Li X, Wang K, Ren Y, Zhang L, Tang XJ, Zhang HM, Zhao CQ, Liu PJ, Zhang JM and He JJ: MAPK signaling mediates sino-menine hydrochloride-induced human breast cancer cell death via both reactive oxygen species-dependent and -independent pathways: An in vitro and in vivo study. Cell Death Dis. 5:e13562014. View Article : Google Scholar
29	Witek ME, Snook AE, Lin JE, Blomain ES, Xiang B, Magee MS and Waldman SA: A novel CDX2 isoform regulates alternative splicing. PLoS One. 9:e1042932014. View Article : Google Scholar : PubMed/NCBI
30	Bae JM, Lee TH, Cho NY, Kim TY and Kang GH: Loss of CDX2 expression is associated with poor prognosis in colorectal cancer patients. World J Gastroenterol. 21:1457–1467. 2015. View Article : Google Scholar : PubMed/NCBI
31	Olsen J, Eiholm S, Kirkeby LT, Espersen ML, Jess P, Gögenür I, Olsen J and Troelsen JT: CDX2 downregulation is associated with poor differentiation and MMR deficiency in colon cancer. Exp Mol Pathol. 100:59–66. 2016. View Article : Google Scholar
32	Dawson H, Koelzer VH, Lukesch AC, Mallaev M, Inderbitzin D, Lugli A and Zlobec I: Loss of Cdx2 expression in primary tumors and lymph node metastases is specific for mismatch repair-deficiency in colorectal cancer. Front Oncol. 3:2652013. View Article : Google Scholar : PubMed/NCBI
33	Olsen J, Espersen ML, Jess P, Kirkeby LT and Troelsen JT: The clinical perspectives of CDX2 expression in colorectal cancer: A qualitative systematic review. Surg Oncol. 23:167–176. 2014. View Article : Google Scholar : PubMed/NCBI
34	Zheng JB, Qiao LN, Sun XJ, Qi J, Ren HL, Wei GB, Zhou PH, Yao JF, Zhang L and Jia PB: Overexpression of caudal-related homeobox transcription factor 2 inhibits the growth of transplanted colorectal tumors in nude mice. Mol Med Rep. 12:3409–3415. 2015.PubMed/NCBI
35	Xu Y, Hou J, Liu Z, Yu H, Sun W, Xiong J, Liao Z, Zhou F, Xie C and Zhou Y: Gene therapy with tumor-specific promoter mediated suicide gene plus IL-12 gene enhanced tumor inhibition and prolonged host survival in a murine model of Lewis lung carcinoma. J Transl Med. 9:392011. View Article : Google Scholar : PubMed/NCBI
36	Zhang Y, Toh L, Lau P and Wang X: Human telomerase reverse transcriptase (hTERT) is a novel target of the Wnt/β-catenin pathway in human cancer. J Biol Chem. 287:32494–32511. 2012. View Article : Google Scholar : PubMed/NCBI
37	Shepelev MV, Kopantzev EP, Vinogradova TV, Sverdlov ED and Korobko IV: hTERT and BIRC5 gene promoters for cancer gene therapy: A comparative study. Oncol Lett. 12:1204–1210. 2016.PubMed/NCBI
38	Paolicchi E, Gemignani F, Krstic-Demonacos M, Dedhar S, Mutti L and Landi S: Targeting hypoxic response for cancer therapy. Oncotarget. 7:13464–13478. 2016.PubMed/NCBI
39	Zhang H, Liang C, Hou X, Wang L and Zhang D: Study of the combined treatment of lung cancer using gene-loaded immunomagnetic albumin nanospheres in vitro and in vivo. Int J Nanomed. 11:1039–1050. 2016. View Article : Google Scholar
40	Harvey TJ, Hennig IM, Shnyder SD, Cooper PA, Ingram N, Hall GD, Selby PJ and Chester JD: Adenovirus-mediated hypoxia-targeted gene therapy using HSV thymidine kinase and bacterial nitroreductase prodrug-activating genes in vitro and in vivo. Cancer Gene Ther. 18:773–784. 2011. View Article : Google Scholar : PubMed/NCBI
41	Wang W, Sun X, Lu L, Zheng JB, Tian Y and Wang W: Cytotoxicity of lymphocytes activated by superantigen toxic-shock-syndrome toxin-1 against colorectal cancer LoVo cells. Mol Cell Biochem. 376:1–9. 2013. View Article : Google Scholar : PubMed/NCBI
42	Brabletz T, Spaderna S, Kolb J, Hlubek F, Faller G, Bruns CJ, Jung A, Nentwich J, Duluc I, Domon-Dell C, et al: Down-regulation of the homeodomain factor Cdx2 in colorectal cancer by collagen type I: An active role for the tumor environment in malignant tumor progression. Cancer Res. 64:6973–6977. 2004. View Article : Google Scholar : PubMed/NCBI
43	Turck N, Gross I, Gendry P, Stutzmann J, Freund JN, Kedinger M, Simon-Assmann P and Launay JF: Laminin isoforms: Biological roles and effects on the intracellular distribution of nuclear proteins in intestinal epithelial cells. Exp Cell Res. 303:494–503. 2005. View Article : Google Scholar : PubMed/NCBI
44	Yusra, Semba S and Yokozaki H: Biological significance of tumor budding at the invasive front of human colorectal carcinoma cells. Int J Oncol. 41:201–210. 2012.PubMed/NCBI
45	Wei W, Li L, Wang X, Yan L, Cao W, Zhan Z, Zhang X, Yu H, Xie Y and Xiao Q: Overexpression of caudal type homeobox transcription factor 2 inhibits the growth of the MGC-803 human gastric cancer cell line in vivo. Mol Med Rep. 12:905–912. 2015.PubMed/NCBI
46	Seno H, Oshima M, Taniguchi MA, Usami K, Ishikawa TO, Chiba T and Taketo MM: CDX2 expression in the stomach with intestinal metaplasia and intestinal-type cancer: Prognostic implications. Int J Oncol. 21:769–774. 2002.PubMed/NCBI