Gene silencing of galectin-3 changes the biological behavior of Eca109 human esophageal cancer cells

Qiao,Lili; Liang,Ning; Xie,Jian; Luo,Hui; Zhang,Jingxin; Deng,Guodong; Li,Yupeng; Zhang,Jiandong

doi:10.3892/mmr.2015.4543

Gene silencing of galectin-3 changes the biological behavior of Eca109 human esophageal cancer cells

Authors:
- Lili Qiao
- Ning Liang
- Jian Xie
- Hui Luo
- Jingxin Zhang
- Guodong Deng
- Yupeng Li
- Jiandong Zhang
View Affiliations

Affiliations: Department of Radiation Oncology, Qianfoshan Hospital, Shandong University, Jinan, Shandong 250014, P.R. China, Graduate School, Weifang Medical College, Weifang, Shandong 261053, P.R. China, Graduate School, Taishan Medical College, Taian, Shandong 271021, P.R. China
Published online on: November 10, 2015 https://doi.org/10.3892/mmr.2015.4543
Pages: 160-166
Copyright: © Qiao 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

Galectin-3 is a multifunctional β-galactoside‑binding lectin that is involved in multiple biological functions which are upregulated in malignancies, including cell growth, adhesion, proliferation, progression and metastasis, as well as apoptosis. A previous study has confirmed the roles of galecin-3 overexpression in the biological behavior of Eca109 human esophageal cancer (EC) cells. In the present study, small interfering (si)RNA-mediated galectin-3 silencing was performed to analyze the effects of decreased galectin-3 expression on the biological behavior of EC cells. Western blot and quantitative polymerase chain reaction analyses were utilized to confirm galectin-3 knockdown at the protein and mRNA level (P<0.05 vs. siRNA-control and untransfected groups). Cell proliferation was assessed using the Cell Counting Kit-8 assay. At 72 and 96 h after transfection, the proliferation of Eca109 cells in the siRNA-Gal-3 group was decreased compared with that in the siRNA-Control and untransfected groups (P<0.001 and P=0.004, respectively). Furthermore, Transwell assays demonstrated that inhibition of galecin-3 significantly reduced the migration and invasion of Eca109 cells compared with that in the other groups (P<0.05). Finally, apoptosis of Eca109 cells was detected using Annexin V/7-amino‑actinomycin double-staining and flow cytometric analysis. Galectin-3 knockdown significantly enhanced the apoptotic rate of Eca109 cells compared with that in the siRNA-control and untreated groups (P=0.031 and P=0.047, respectively). In conclusion, following successful knockdown of galecin-3 expression in Eca109 cells, the cell proliferation, migration and invasion were reduced, while the apoptosis was enhanced, which indicates that galectin silencing may represent a therapeutic strategy for EC.

View Figures

View References

1	Dumic J, Dabelic S and Flögel M: Galectin-3: An open-ended story. Biochim Biophys Acta. 1760:616–635. 2006. View Article : Google Scholar : PubMed/NCBI
2	Barondes SH, Castronovo V, Cooper DN, Cummings RD, Drickamer K, Feizi T, Gitt MA, Hirabayashi J, Hughes C, Kasai K, et al: Galectins: A family of animal β-galactoside-binding lectins. Cell. 76:597–598. 1994. View Article : Google Scholar : PubMed/NCBI
3	Ochieng J, Fridman R, Nangia-Makker P, Kleiner DE, Liotta LA, Stetler-Stevenson WG and Raz A: Galectin-3 is a novel substrate for human matrix metalloproteinases-2 and -9. Biochemistry. 33:14109–14114. 1994. View Article : Google Scholar : PubMed/NCBI
4	Gong HC, Honjo Y, Nangia-Makker P, Hogan V, Mazurak N, Bresalier RS and Raz A: The NH2 terminus of galectin-3 governs cellular compartmentalization and functions in cancer cells. Cancer Res. 59:6239–6245. 1999.
5	Zhang HY, Jin L, Stilling GA, Ruebel KH, Coonse K, Tanizaki Y, Raz A and Lloyd RV: RUNX1 and RUNX2 upregulate Galectin-3 expression in human pituitary tumors. Endocrine. 35:101–111. 2009. View Article : Google Scholar
6	Sakaki M, Fukumori T, Fukawa T, Elsamman E, Shiirevnyamba A, Nakatsuji H and Kanayama HO: Clinical significance of Galectin-3 in clear cell renal cell carcinoma. J Med Invest. 57:152–157. 2010. View Article : Google Scholar : PubMed/NCBI
7	Jia W, Kidoya H, Yamakawa D, Naito H and Takakura N: Galectin-3 accelerates M2 macrophage infiltration and angio-genesis in tumors. Am J Pathol. 182:1821–1831. 2013. View Article : Google Scholar : PubMed/NCBI
8	Wu SW, Yu L, Zhou L, Cheng ZN and Tao YS: Expression of Gal-3 and CD82/KAI1 proteins in non-small cell lung cancer and their clinical significance. Chinese Journal of Oncology. 35:124–128. 2013.In Chinese.
9	Ochieng J, Furtak V and Lukyanov P: Extracellular functions of galectin-3. Glycoconj J. 19:527–535. 2004. View Article : Google Scholar : PubMed/NCBI
10	Yu F, Finley RL Jr, Raz A and Kim HR: Galectin-3 translocates to the perinuclear membranes and inhibits cytochrome c release from the mitochondria. A role for synexin in galectin-3 translocation. J Biol Chem. 277:15819–15827. 2002. View Article : Google Scholar : PubMed/NCBI
11	Takenaka Y, Fukumori T, Yoshii T, Oka N, Inohara H, Kim HR, Bresalier RS and Raz A: Nuclear export of phosphorylated galectin-3 regulates its antiapoptotic activity in response to chemotherapeutic drugs. Mol Cell Biol. 24:4395–4406. 2004. View Article : Google Scholar : PubMed/NCBI
12	Patterson RJ, Wang W and Wang JL: Understanding the biochemical activities of galectin-1 and galectin-3 in the nucleus. Glycoconj J. 19:499–506. 2004. View Article : Google Scholar : PubMed/NCBI
13	Shimura T, Takenaka Y, Tsutsumi S, Hogan V, Kikuchi A and Raz A: Galectin-3, a novel binding partner of beta-catenin. Cancer Res. 64:6363–6367. 2004. View Article : Google Scholar : PubMed/NCBI
14	Braeuer RR, Shoshan E, Kamiya T and Bar-Eli M: The sweet and bitter sides of galectins in melanoma progression. Pigment Cell Melanoma Res. 25:592–601. 2012. View Article : Google Scholar : PubMed/NCBI
15	Lotan R, Ito H, Yasui W, Yokozaki H, Lotan D and Tahara E: Expression of a 31-kDa lactose-binding lectin in normal human gastric mucosa and in primary and metastatic gastric carcinomas. Int J Cancer. 56:474–480. 1994. View Article : Google Scholar : PubMed/NCBI
16	Inohara H, Honjo Y, Yoshii T, Akahani S, Yoshida J, Hattori K, Okamoto S, Sawada T, Raz A and Kubo T: Expression of galectin-3 in fine-needle aspirates as a diagnostic marker differentiating benign from malignant thyroid neoplasms. Cancer. 85:2475–2484. 1999. View Article : Google Scholar : PubMed/NCBI
17	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
18	Rice TW, Rusch VW, Apperson-Hansen C, Allen MS, Chen LQ, Hunter JG, Kesler KA, Law S, Lerut TE, Reed CE, et al: Worldwide esophageal cancer collaboration. Dis Esophagus. 22:1–8. 2009. View Article : Google Scholar : PubMed/NCBI
19	Liang N, Song X, Xie J, Xu D, Liu F, Yu X, Tian Y, Liu Z, Qiao L and Zhang J: Effect of galectin-3 on the behavior of Eca-109 human esophageal cancer cells. Mol Med Rep. 11:896–902. 2015.
20	Du YY, Zhao LM, Chen L, Sang MX, Li J, Ma M and Liu JF: The tumor-suppressive function of miR-1 by targeting LASP1 and TAGLN2 in esophageal squamous cell carcinoma. J Gastroenterol Hepatol. Epub ahead of print. 2015. View Article : Google Scholar
21	Li S, Qin X, Li Y, Zhang X, Niu R, Zhang H, Cui A, An W and Wang X: MiR-133a suppresses the migration and invasion of esophageal cancer cells by targeting the EMT regulator SOX4. Am J Transl Res. 7:1390–1403. 2015.PubMed/NCBI
22	Zhou W, Yue H, Li C, Chen H and Yuan Y: Protein arginine methyltransferase 1 promoted the growth and migration of cancer cells in esophageal squamous cell carcinoma. Tumour Biol. 2015.
23	Pei Y, Wang P, Liu H, He F and Ming L: FOXQ1 promotes esophageal cancer proliferation and metastasis by negatively modulating CDH1. Biomed Pharmacother. 74:89–94. 2015. View Article : Google Scholar : PubMed/NCBI
24	Li J, Zhu SC, Li SG, Zhao Y, Xu JR and Song CY: TKTL1 promotes cell proliferation and metastasis in esophageal squamous cell carcinoma. Biomed Pharmacother. 74:71–76. 2015. View Article : Google Scholar : PubMed/NCBI
25	Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29:e452001. View Article : Google Scholar : PubMed/NCBI
26	Hood JD and Cheresh DA: Role of integrins in cell invasion and migration. Nat Rev Cancer. 2:91–100. 2002. View Article : Google Scholar
27	Liu FT and Rabinovich GA: Galectins as modulators of tumour progression. Nat Rev Cancer. 5:29–41. 2005. View Article : Google Scholar : PubMed/NCBI
28	Kobayashi T, Shimura T, Yajima T, Kubo N, Araki K, Tsutsumi S, Suzuki H, Kuwano H and Raz A: Transient gene silencing of galectin-3 suppresses pancreatic cancer cell migration and invasion through degradation of β-catenin. Int J Cancer. 129:2775–2786. 2011. View Article : Google Scholar : PubMed/NCBI
29	Honjo Y, Nangia-Makker P, Inohara H and Raz A: Down-regulation of Galectin-3 suppresses tumorigenicity of human breast carcinoma cells. Clin Cancer Res. 7:661–668. 2001.PubMed/NCBI
30	Bresalier RS, Mazurek N, Sternberg LR, Byrd JC, Yunker CK, Nangia-Makker P and Raz A: Metastasis of human colon cancer is altered by modifying expression of the beta-galactoside-binding protein galectin-3. Gastroenterology. 115:287–296. 1998. View Article : Google Scholar : PubMed/NCBI
31	Hittelet A, Camby I, Nagy N, Legendre H, Bronckart Y, Decaestecker C, Kaltner H, Nifant'ev NE, Bovin NV, Pector JC, et al: Binding sites for lewis antigens are expressed by human colon cancer cells and negatively affect their migration. Lab Invest. 83:777–787. 2003. View Article : Google Scholar : PubMed/NCBI
32	Debray C, Vereecken P, Belot N, Teillard P, Brion JP, Pandolfo M and Pochet R: Multifaceted role of galectin-3 on human glioblastoma cell motility. Biochem Biophys Res Commun. 325:1393–1398. 2004. View Article : Google Scholar : PubMed/NCBI
33	Glinsky VV, Glinsky GV, Glinskii OV, Huxley VH, Turk JR, Mossine VV, Deutscher SL, Pienta KJ and Quinn TP: Intravascular metastatic cancer cell homotypic aggregation at the sites of primary attachment to the endothelium. Cancer Res. 63:3805–3811. 2003.PubMed/NCBI
34	O'Driscoll L, Linehan R, Liang YH, Joyce H, Oglesby I and Clynes M: Galectin-3 expression alters adhesion, motility and invasion in a lung cell line (DLKP), in vitro. Anticancer Res. 22:3117–3125. 2002.
35	Junking M, Wongkham C, Sripa B, Sawanyawisuth K, Araki N and Wongkham S: Decreased expression of galectin-3 is associated with metastatic potential of liver fluke-associated cholangiocarcinoma. Eur J Cancer. 44:619–626. 2008. View Article : Google Scholar : PubMed/NCBI
36	Endo K, Kohnoe S, Tsujita E, Watanabe A, Nakashima H, Baba H and Maehara Y: Galectin-3 expression is a potent prognostic marker in colorectal cancer. Anticancer Res. 25:3117–3121. 2005.PubMed/NCBI
37	Shi Y, He B, Kuchenbecker KM, You L, Xu Z, Mikami I, Yagui-Beltran A, Clement G, Lin YC, Okamoto J, et al: Inhibition of Wnt-2 and galectin-3 synergistically destabilizes beta-catenin and induces apoptosis in human colorectal cancer cells. Int J Cancer. 121:1175–1181. 2007. View Article : Google Scholar : PubMed/NCBI
38	Zaia Povegliano L, Oshima CT, de Oliveira Lima F, Andrade Scherholz PL and Manoukian Forones N: Immunoexpression of galectin-3 in colorectal cancer and its relationship with survival. J Gastrointest Cancer. 42:217–221. 2011. View Article : Google Scholar
39	Lotz MM, Andrews CW Jr, Korzelius CA, Lee EC, Steele GD Jr, Clarke A and Mercurio AM: Decreased expression of Mac-2 (carbohydrate binding protein 35) and loss of its nuclear localization are associated with the neoplastic progression of colon carcinoma. Proc Natl Acad Sci USA. 90:3466–3470. 1993. View Article : Google Scholar : PubMed/NCBI
40	Liu L, Sakai T, Sano N and Fukui K: Nucling mediates apoptosis by inhibiting expression of galectin-3 through interference with nuclear factor kappaB signaling. Biochem J. 380:31–41. 2004. View Article : Google Scholar : PubMed/NCBI
41	Dumic J, Lauc G and Flögel M: Expression of galectin-3 in cells exposed to stress-roles of jun and NF-kappaB. Cell Physiol Biochem. 10:149–158. 2000. View Article : Google Scholar : PubMed/NCBI
42	Baichwal VR and Baeuerle PA: Activate NF-kappaB or die? Curr Biol. 7:R94–R96. 1997. View Article : Google Scholar : PubMed/NCBI
43	Baeuerle PA and Henkel T: Function and activation of NF-kappaB in the immune system. Annu Rev Immunol. 12:141–179. 1994. View Article : Google Scholar
44	Lin CI, Whang EE, Abramson MA, Donner DB, Bertagnolli MM, Moore FD Jr and Ruan DT: Galectin-3 regulates apoptosis and doxorubicin chemoresistance in papillary thyroid cancer cells. Biochem Biophys Res Commun. 379:626–631. 2009. View Article : Google Scholar : PubMed/NCBI
45	Oka N, Nakahara S, Takenaka Y, Fukumori T, Hogan V, Kanayama HO, Yanagawa T and Raz A: Galectin-3 inhibits tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by activating Akt in human bladder carcinoma cells. Cancer Res. 65:7546–7553. 2005.PubMed/NCBI
46	Choi JH, Chun KH, Raz A and Lotan R: Inhibition of N-(4-hydroxyphenyl) retinamide-induced apoptosis in breast cancer cells by galectin-3. Cancer Biol Ther. 3:447–452. 2004. View Article : Google Scholar : PubMed/NCBI
47	Tepsiri N, Chaturat L, Sripa B, Namwat W, Wongkham S, Bhudhisawasdi V and Tassaneeyakul W: Drug sensitivity and drug resistance profiles of human intrahepatic cholangiocar-cinoma cell lines. World J Gastroenterol. 11:2748–2753. 2005. View Article : Google Scholar : PubMed/NCBI