J Gastric Cancer.  2012 Jun;12(2):73-80.

Amplification of the UQCRFS1 Gene in Gastric Cancers

• Jun KH
• Kim SY
• Yoon JH
• Song JH
• Park WS

Abstract

PURPOSE
The specific aim of this study is to unravel a DNA copy number alterations, and to search for novel genes that are associated with the development of Korean gastric cancer.
MATERIALS AND METHODS
We investigated a DNA copy number changes in 23 gastric adenocarcinomas by array-comparative genomic hybridization and quantitative real-time polymerase chain reaction analyses. Besides, the expression of UQCRFS1, which shows amplification in array-CGH, was examined in 186 gastric cancer tissues by an immunohistochemistry, and in 9 gastric cancer cell lines, as well as 24 gastric cancer tissues by immunoblotting.
RESULTS
We found common gains at 48 different loci, and a common loss at 19 different loci. Amplification of UQCRFS1 gene at 19q12 was found in 5 (21.7%) of the 23 gastric cancers in an array-comparative genomic hybridization and DNA copy number were increased in 5 (20.0%) out of the 25 gastric cancer in quantitative real-time polymerase chain reaction. In immunohistochemistry, the overexpression of the protein was detected in 105 (56.5%) out of the 186 gastric cancer tissues. Statistically, there was no significant relationship between the overexpression of UQCRFS1 and clinicopathologic parameters (P>0.05). In parallel, the overexpression of UQCRFS1 protein was confirmed in 6 (66.7%) of the 9 gastric cancer cell lines, and 12 (50.0%) of the 24 gastric cancer tissues by immunoblotting.
CONCLUSIONS
These results suggest that the overexpression of UQCRFS1 gene may contribute to the development and/or progression of gastric cancer, and further supported that mitochondrial change may serve as a potential cancer biomarker.

Keyword

Stomach neoplasms; ArrayCGH; Immunohistochemistry; Amplification; Overexpression

MeSH Terms

Adenocarcinoma
Cell Line
Coat Protein Complex I
DNA
DNA Copy Number Variations
Immunohistochemistry
Nucleic Acid Hybridization
Real-Time Polymerase Chain Reaction
Stomach Neoplasms
Coat Protein Complex I
DNA

Figure

• 

  Fig. 1 Array-comparative genome hybridization result showing amplification of chromosome 19q13 locus, which the UQCRFS1 gene is located, in gastric cancers.

• 

  Fig. 2 Copy number change of UQCRFS1 in gastric cancers. In real time polymerase chain reaction, copy number ratio was calculated based on the corresponding normal value of each case. UQCRFS1 copy number was increased more than 1.5 fold in 5 (20%) of 25 gastric cancer DNAs compared to the surrounding gastric mucosal tissue DNAs. Cont = control; T = tumor.

• 

  Fig. 3 Immunostaining of the UQCRFS1 protein in gastric cancer. Expression of the UQCRFS1 protein in gastric cancers. Low-power view (×2, magnification) of tissue cores stained for UQCRFS1. Immunostaining for UQCRFS1 was detected in cytoplasm and nucleus of non-cancerous gastric mucosal epithelial cells (×200, magnification) (A, B). Overexpression of UQCRFS1 protein was seen in well differentiated adenocarcinoma (C) and moderate differentiated adenocarcinoma (D) (×200, magnification).

• 

  Fig. 4 Immunoblotting analysis of UQCRFS1 protein expression in gastric cancer cell lines and tissues. Total cell lysates prepared from different cell lines and tissues were subjected to immunoblotting analysis with UQCRFS1, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and actin antibodies. Compared with UQCRFS1 expression in corresponding con-cancerous gastric mucosa, expression of the protein was increased in 6 (66.7%) of 9 gastric cancer cell lines and 12 (50.0%) of 24 gastric cancer tissues, respectively.

Reference

1. Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, et al. Cancer statistics, 2005. CA Cancer J Clin. 2005. 55:10–30.

2. Jung KW, Park S, Kong HJ, Won YJ, Lee JY, Park EC, et al. Cancer statistics in Korea: incidence, mortality, survival, and prevalence in 2008. Cancer Res Treat. 2011. 43:1–11.

3. Snijders AM, Nowak N, Segraves R, Blackwood S, Brown N, Conroy J, et al. Assembly of microarrays for genome-wide measurement of DNA copy number. Nat Genet. 2001. 29:263–264.

4. Buffart TE, Carvalho B, Hopmans E, Brehm V, Kranenbarg EK, Schaaij-Visser TB, et al. Gastric cancers in young and elderly patients show different genomic profiles. J Pathol. 2007. 211:45–51.

5. Weiss MM, Kuipers EJ, Postma C, Snijders AM, Pinkel D, Meuwissen SG, et al. Genomic alterations in primary gastric adenocarcinomas correlate with clinicopathological characteristics and survival. Cell Oncol. 2004. 26:307–317.

6. Weiss MM, Kuipers EJ, Postma C, Snijders AM, Siccama I, Pinkel D, et al. Genomic profiling of gastric cancer predicts lymph node status and survival. Oncogene. 2003. 22:1872–1879.

7. Sugihara H, Hattori T, Fujita S, Hirose K, Fukuda M. Regional ploidy variations in signet ring cell carcinomas of the stomach. Cancer. 1990. 65:122–129.

8. Hattori T, Sugihara H, Fukuda M, Hamada S, Fujita S. DNA ploidy patterns of minute carcinomas in the stomach. Jpn J Cancer Res. 1986. 77:276–281.

9. Rozen S, Skaletsky H. Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 2000. 132:365–386.

10. Kononen J, Bubendorf L, Kallioniemi A, Bärlund M, Schraml P, Leighton S, et al. Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med. 1998. 4:844–847.

11. Yoon JH, Song JH, Zhang C, Jin M, Kang YH, Nam SW, et al. Inactivation of the Gastrokine 1 gene in gastric adenomas and carcinomas. J Pathol. 2011. 223:618–625.

12. Tsukamoto Y, Uchida T, Karnan S, Noguchi T, Nguyen LT, Tanigawa M, et al. Genome-wide analysis of DNA copy number alterations and gene expression in gastric cancer. J Pathol. 2008. 216:471–482.

13. Rossi E, Klersy C, Manca R, Zuffardi O, Solcia E. Correlation between genomic alterations assessed by array comparative genomic hybridization, prognostically informative histologic subtype, stage, and patient survival in gastric cancer. Hum Pathol. 2011. 42:1937–1945.

14. Bizari L, Borim AA, Leite KR, Gonçalves Fde T, Cury PM, Tajara EH, et al. Alterations of the CCND1 and HER-2/neu (ERBB2) proteins in esophageal and gastric cancers. Cancer Genet Cytogenet. 2006. 165:41–50.

15. Calcagno DQ, Leal MF, Seabra AD, Khayat AS, Chen ES, Demachki S, et al. Interrelationship between chromosome 8 aneuploidy, C-MYC amplification and increased expression in individuals from northern Brazil with gastric adenocarcinoma. World J Gastroenterol. 2006. 12:6207–6211.

16. Wolf M, Mousses S, Hautaniemi S, Karhu R, Huusko P, Allinen M, et al. High-resolution analysis of gene copy number alterations in human prostate cancer using CGH on cDNA microarrays: impact of copy number on gene expression. Neoplasia. 2004. 6:240–247.

17. Hyman E, Kauraniemi P, Hautaniemi S, Wolf M, Mousses S, Rozenblum E, et al. Impact of DNA amplification on gene expression patterns in breast cancer. Cancer Res. 2002. 62:6240–6245.

18. Järvinen AK, Autio R, Haapa-Paananen S, Wolf M, Saarela M, Grénman R, et al. Identification of target genes in laryngeal squamous cell carcinoma by high-resolution copy number and gene expression microarray analyses. Oncogene. 2006. 25:6997–7008.

19. Kim HK, Park WS, Kang SH, Warda M, Kim N, Ko JH, et al. Mitochondrial alterations in human gastric carcinoma cell line. Am J Physiol Cell Physiol. 2007. 293:C761–C771.

20. Ohashi Y, Kaneko SJ, Cupples TE, Young SR. Ubiquinol cytochrome c reductase (UQCRFS1) gene amplification in primary breast cancer core biopsy samples. Gynecol Oncol. 2004. 93:54–58.

21. Sait SN, Qadir MU, Conroy JM, Matsui S, Nowak NJ, Baer MR. Double minute chromosomes in acute myeloid leukemia and myelodysplastic syndrome: identification of new amplification regions by fluorescence in situ hybridization and spectral karyotyping. Genes Chromosomes Cancer. 2002. 34:42–47.

22. Kaneko SJ, Gerasimova T, Smith ST, Lloyd KO, Suzumori K, Young SR. CA125 and UQCRFS1 FISH studies of ovarian carcinoma. Gynecol Oncol. 2003. 90:29–36.

23. Rieske JS. Composition, structure, and function of complex III of the respiratory chain. Biochim Biophys Acta. 1976. 456:195–247.

24. Trumpower BL, Edwards CA. Purification of a reconstitutively active iron-sulfur protein (oxidation factor) from succinate. cytochrome c reductase complex of bovine heart mitochondria. J Biol Chem. 1979. 254:8697–8706.

25. Fiskum G, Starkov A, Polster BM, Chinopoulos C. Mitochondrial mechanisms of neural cell death and neuroprotective interventions in Parkinson's disease. Ann N Y Acad Sci. 2003. 991:111–119.

26. Owens KM, Kulawiec M, Desouki MM, Vanniarajan A, Singh KK. Impaired OXPHOS complex III in breast cancer. PLoS One. 2011. 6:e23846.