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The tumor supressor function of STGC3 and its reduced expression in nasopharyngeal carcinoma

Abstract

STGC3 is a novel candidate tumor suppressor gene that was found to be associated with nasopharyngeal carcinoma (NPC) via the cDNA cloning and RACE processes. The biological function of the STGC3 protein and its expression level in nasopharyngeal carcinoma remain unknown. This study aimed to evaluate the STGC3 protein expression level in NPC and to investigate the inhibitory function of STGC3 as a candidate tumor suppressor gene. We assessed the expression of the STGC3 protein in NPC biopsies and normal control specimens via Western blot and immunohistochemical analysis. The expression of STGC3 as induced by doxycycline (Dox) via a tetracycline (Tet)-regulated system in human nasopharyngeal carcinoma cell line CNE2 was also established, and the effect of STGC3 restoration on the biological behavior of CNE2 was observed. A reduced level of STGC3 expression (0.978 ± 0.213 versus 0.324 ± 0.185, P < 0.05) was detected in NPC versus normal nasopharyngeal tissue by Western blot assay. Immunohistochemical assays for STGC3 detected positive staining in the nuclei and cytoplasm of epithelial cells, and the positive expression rate in NPC, 8 of 21 (38%), was lower than that in normal nasopharynx samples, 16 of 22 (72%). After STGC3 expression was restored, the growth capacity and clone formation potential of CNE2 cells in soft agar were significantly suppressed, and the cell percentage in G0/G1 phase increased, while the percentage of cells entering the S and G2 phases decreased. This indicates that an abnormality in STGC3 expression is associated with nasopharyngeal carcinogenesis and that it may play an important role in controlling cell growth and regulating the cell cycle.

Abbreviations

AMV RT:

AMV reverse transcriptase

ANOVA:

one-way analysis of variance

DAB:

diaminobenzidine

Dox:

doxycycline

FCM:

flow cytometry

IOD:

integral optical densities

NPC:

nasopharyngeal carcinoma

PBS:

phosphate-buffered saline

PI:

propidium iodide

PVDF:

polyvinylidene difluoride

RACE:

Rapid Amplification of cDNA End

rtTA:

reverse tetracycline/doxycycline-responsive transcriptional activator

S-P:

streptavidin-peroxidase

Tet:

tetracycline

References

  1. Ho, J.H.C. An epidemiologic and clinical study of nasopharyngeal carcinoma. Int. Radiat. Oncol. Biol. Phys. 4 (1978) 183–205.

    Google Scholar 

  2. Zheng, Y.M., Tuppin, P. and Hubert, A. The environmental and dietary risk factors for nasopharyngeal carcinoma: a case-control study in Zangwu County, Guangxi, China. Br. J. Cancer 69 (1994) 508–514.

    PubMed  CAS  Google Scholar 

  3. Ward, M.H., Pan, W.H., Cheng, Y.J., Li, F.H., Brinton, L.A., Chen, C.J., Hsu, M.M., Chen, I.H., Levine, P.H., Yang, C.S. and Hildesheim, A. Dietary exposure to nitrite and nitrosamines and risk of nasopharyngeal carcinoma in Taiwan. Int. J. Cancer 86 (2000) 603–609.

    Article  PubMed  CAS  Google Scholar 

  4. Krishna, S.M., James, S., Kattoor, J. and Balaram, P. Serum EBV DNA as a biomarker in primary nasopharyngeal carcinoma of Indian origin. Jpn. J._Clin. Oncol. 34 (2004) 307–311.

    Article  PubMed  Google Scholar 

  5. Yang, X.R., Diehl, S., Pfeiffer, R., Chen, C.J., Hsu, W.L., Dosemeci, M., Cheng, Y.J., Sun, B., Goldstein, A.M. and Hildesheim, A. Chinese and American Genetic Epidemiology of NPC Study Team. Evaluation of risk factors for nasopharyngeal carcinoma in high-risk nasopharyngeal carcinoma families in Taiwan. Cancer Epidemiol. Biomarkers Prev. 14 (2005) 900–905.

    Article  PubMed  CAS  Google Scholar 

  6. Thornburg, N.J., Pathmanathan, R. and Raab-Traub, N. Activation of nuclear factor-kappaB p50 homodimer/Bcl-3 complexes in nasopharyngeal carcinoma. Cancer Res. 63 (2003) 8293–8301.

    PubMed  CAS  Google Scholar 

  7. Horikawa, T., Sheen, T.S., Takeshita, H., Sato, H., Furukawa, M. and Yoshizaki, T. Induction of c-Met proto-oncogene by Epstein-Barr virus latent membrane protein-1 and the correlation with cervical lymph node metastasis of nasopharyngeal carcinoma. Am. J. Pathol. 159 (2001) 27–33.

    PubMed  CAS  Google Scholar 

  8. Sarac, S., Akyol, M.U., Kanbur, B., Poyraz, A., Akyol, G., Yilmaz, T. and Sungur, A. Bcl-2 and LMP1 expression in nasopharyngeal carcinomas. Am. J. Otolaryngol. 22 (2001) 377–382.

    Article  PubMed  CAS  Google Scholar 

  9. Qian, C.N., Guo, X., Cao, B., Kort, E.J., Lee, C.C., Chen, J., Wang, L.M., Mai, W.Y., Min, H.Q., Hong, M.H., Vande, Woude, G.F., Resau, J.H. and The, B.T. Met protein expression level correlates with survival in patients with late-stage nasopharyngea carcinoma. Cancer Res. 62 (2002) 589–596.

    PubMed  CAS  Google Scholar 

  10. Lo, K.W., Cheung, S.T., Leung, S.F., van Hasselt, A., Tsang, Y.S., Mak, K.F., Chung, Y.F., Woo, J.K., Lee, J.C. and Huang, D.P. Hypermethylation of the p16 gene in nasopharyngeal carcinoma. Cancer Res. 56 (1996) 2721–2725.

    PubMed  CAS  Google Scholar 

  11. Yan, J., Fang, Y., Huang, B.J., Liang, Q.W., Wu, Q.L. and Zeng, Y.X. Absence of evidence for HER2 amplication in nasopharyngeal carcinoma. Cancer Genet. Cytogenet. 132 (2002) 116–119.

    Article  PubMed  CAS  Google Scholar 

  12. Sheu, L.F., Chen, A., Tseng, H.H., Leu, F.J., Lin, J.K., Ho, K.C. and Meng, C.L. Assessment of p53 expression in nasopharyngeal carcinoma. Hum. Pathol. 26 (1995) 380–386.

    Article  PubMed  CAS  Google Scholar 

  13. Hui, A.B., Lo, K.W., Kwong, J., Lam, E.C., Chan, S.Y., Chow, L.S., Chan, A.S., Teo, P.M. and Huang, D.P. Epigenetic inactivation of TSLC1 gene in nasopharyngeal carcinoma. Mol. Carcinog. 38 (2003) 170–178.

    Article  PubMed  CAS  Google Scholar 

  14. Shao, J.Y., Wang, H.Y., Huang, X.M., Feng, Q.S., Huang, P., Feng, B.J., Huang, L.X., Yu, X.J., Li, J.T., Hu, L.F., Ernberg, I. and Yi, X.Z. Genomewide allelotype analysis of sporadic primary nasopharyngeal carcinoma from southern China. Int. J. Oncol. 17 (2000) 1267–1275.

    PubMed  CAS  Google Scholar 

  15. Deng, L., Jing, N., Tan, G., Zhou, M., Zhan, F., Xie, Y., Cao, L. and Li, G. A common region of allelic loss on chromosome region3p25.3-26.3 in nasopharyngeal carcinoma. Genes Chromosomes Cancer 23 (1998) 21–22.

    Article  PubMed  CAS  Google Scholar 

  16. Shao, J., Li, Y., Wu, Q., Liang, X., Yu, X., Huang, L., Hou, J., Huang, X., Ernberg, I., Hu, L.F. and Zeng, Y. High frequency loss of heterozygosity on the long arms of chromosomes 13 and 14 in nasopharyngeal carcinoma in Southern China. Chin. Med. J. 115 (2002) 571–575.

    PubMed  CAS  Google Scholar 

  17. Xiong, W., Zeng, Z.Y., Xia, J.H., Xia, K., Shen, S.R., Li, X.L., Hu, D.X., Tan, C., Xiang, J.J., Zhou, J., Deng, H., Fan, S.Q., Li, W.F., Wang J.R., Zhou, M., Zhu, S.G., Lu, H.B., Qian, J., Zhang, B.C., Wang, J.R., Ma, J., Xiao, B.Y., Huang, H., Zhang, Q.H., Zhou, Y.H., Luo, X.M., Zhou, H.D., Yang, Y.X., Dai, H.P., Feng, G.Y., Pan, Q., Wu, L.Q., He, L. and Li, G.Y. A susceptibility locus at chromosome 3p21 linked to familial nasopharyngeal carcinoma. Cancer Res. 64 (2004) 1972–1974.

    Article  PubMed  CAS  Google Scholar 

  18. He, X.S., Chen, Z.C., Tian, F., Xiao, Z.Q., He, Z.M., Guan, Y.J., Li, F., He, C.M. and Yuan, J.H. Identification of down-regulated expressed sequence tag at chromosome 3p21 in nasopharyngeal carcinoma. Ai Zheng. 22 (2003) 1–5.

    PubMed  Google Scholar 

  19. He, X.S., Xiao, Z.Q., Chen, Z.C., Zhao, S.P., Zhu, J.H., He, Z.M., Li, Y.J., Tian, F. and Yu, Y.H. Molecular cloning and functional analysis of STGC3-a novel gene on chromosome 3p21. Ai Zheng. 23 (2004) 1110–1115.

    PubMed  CAS  Google Scholar 

  20. Andac, Z., Sasaki, T., Mann, K., Bracaccio, A., Deutzman, R. and Timpl, R. Analysis of heparin, alpha-dystroglycan and sulfatide binding to the G domain of the laminin alpha1 chain by site-directed mutagenesis. J. Mol. Biol. 287 (1999) 253–264.

    Article  PubMed  CAS  Google Scholar 

  21. Talts, J.F., Andac, Z., Gohring, W., Brancaccio, A. and Timpl, R. Binding of the G domains of laminin alpha1 and alpha2 chains and perlecan to heparin, sulfatides, alpha-dystroglycan and several extracellular matrix proteins. EMBO. J. 18 (1999) 863–870.

    Article  PubMed  CAS  Google Scholar 

  22. Talts, J.F. and Timpl, R. Mutation of a basic sequence in the laminin alpha2LG3 module leads to a lack of proteolytic processing and has different effects on beta1 integrin-mediated cell adhesion and alphadystroglycan binding. FEBS. Lett. 458 (1999) 319–323.

    Article  PubMed  CAS  Google Scholar 

  23. Yurchenco, P.D., Sung, U., Ward, M.D., Yamada, Y. and O’Rear, J.J. Recombinant laminin G domain mediates myoblast adhesion and heparin binding. J. Biol. Chem. 268 (1993) 8356–8365.

    PubMed  CAS  Google Scholar 

  24. Sung, U., O’Rear, J.J. and Yurchenco, P.D. Localization of heparin binding activity in recombinant laminin G domain. Eur. J. Biochem. 250 (1997) 138–143.

    Article  PubMed  CAS  Google Scholar 

  25. Sambrook, J., Fritsch, E.F. and Maniatis, T. Molecular Cloning. A Laboratory Manual, Ed. I. Cold Spring Harbor Laboratory Press, New York, 1989.

    Google Scholar 

  26. Gossen, M. and Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. 89 (1992) 5547–5551.

    Article  PubMed  CAS  Google Scholar 

  27. Gossen, M., Freundlieb, S., Bender, G., Muller, G., Hillen, W. and Bujard, H. Transcriptional activation by tetracyclines in mammalian cells. Science 268 (1995) 1766–17699.

    Article  PubMed  CAS  Google Scholar 

  28. Kashima, Y., Minami, K. and Seino, S. Establishment of a Tet-on gene expression system in glucose-responsive and-unresponsive MIN6 cells. Diabetes 50 (2001) S133.

    Article  PubMed  CAS  Google Scholar 

  29. Wang, D., Grammer, J.R., Cobbs, C.S., Stewart, J.E. Jr., Liu Z., Rhoden, R., Hecker, T.P., Ding, Q. and Gladson, C.L. P125 focal adhesion kinase promotes malignant astrocytoma cell proliferation in vivo. J. Cell Sci. 113 (2000) 4221–4230.

    PubMed  CAS  Google Scholar 

  30. Kobayashi, T., Sawa, H., Morikawa, J., Zhang, W. and Shiku, H. Bax induction activates apoptotic cascade via mitochondrial cytochrome crelease and Bax overexpression enhances apoptosis induced by chemotherapeutic agents in DLD-1 colon cancer cells. JPN. J. Cancer Res. 91 (2000) 1264–1268.

    PubMed  CAS  Google Scholar 

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Correspondence to Xiu-Sheng He or Zhu-Chu Chen.

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He, XS., Deng, M., Yang, S. et al. The tumor supressor function of STGC3 and its reduced expression in nasopharyngeal carcinoma. Cell Mol Biol Lett 13, 339–352 (2008). https://doi.org/10.2478/s11658-008-0006-9

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  • DOI: https://doi.org/10.2478/s11658-008-0006-9

Keywords

  • Nasopharyngeal neoplasm
  • STGC3
  • Gene expression
  • Tet-on system