Abstract
Real-time PCR was used to quantitate structural and functional aberrations in the tumor suppressor gene RBSP3 (3p.21) in papillomavirus-positive cervical carcinoma (CC). RBSP3 deletions were found in 19 of 45 tumor specimens (42%). The frequency of deletions was reliably higher in tumors with metastases to regional lymph nodes (64%) than in tumors without metastases (32%, P < 0.05). In several cases, RBSP3 amplification was detected. Changes in RBSP3 copy number were observed in 51% (23/45) of the tumor specimens. The RBSP3 transcription level was decreased in 21 of 33 tumors (64%), this event being detected more frequently in tumors with (83%) than without metastases (52%, P < 0.05). In some tumors, RBSP3 mRNA levels were increased. Changes in RBSP3 mRNA level were observed in 79% (26/33) of the CCs. Comparative analysis of the data demonstrated that a decrease in RBSP3 mRNA level was a result of deletions only in 23% of the specimens, while it was not associated with any changes in the gene copy number in 36% of the cases. More unusual combinations of events were an unchanged or even increased mRNA level in the presence of deletions or an increased mRNA level in the absence of changes in gene copy number. It was assumed that deregulation of RBSP3 expression is associated with the progression of CC, that there is more than one mechanism of RBSP3 inactivation, and that RBSP3 has other functions in addition to pRb dephosphorylation.
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References
Zur Hausen H. 2002. Papillomaviruses and cancer: From basic studies to clinical application. Nature Rev. Cancer. 2, 342–350.
Zabarovsky E.R., Lerman M.I., Minna J.D. 2002. Tumor suppressor genes on chromosome 3p involved in the pathogenesis of lung and other cancers. Oncogene. 21, 6915–6935.
Senchenko V., Liu J., Braga E., Mazurenko N., Loginov W., Seryogin Y., Bazov I., Protopopov A., Kisseljov F.L., Kashuba V., Lerman M.I., Klein G., Zabarovsky E.R. 2003. Deletion mapping of cervical carcinomas using quantitative real-time PCR identifies two frequently affected regions in 3p21.3. Oncogene. 22, 2984–2992.
Chatterjee A., Pulido H., Koul S., Beleno N., Perilla A., Posso H., Manusukhani M., Murty V.S. 2001. Mapping the sites of putative suppressor genes at 6p25 and 6p21.3 in cervical carcinoma: Occurrence of allelic deletions in precancerous lesions. Cancer Res. 61, 2119–2123.
Mazurenko N.N., Beliakov I.S., Bliyev A.Yu., Guo Z., Hu X., Vinokourova S.V., Bidzieva B.A., Pavlova L.S., Ponten J., Kisseljov F.L. 2003. Genetic alterations at chromosome 6 associated with cervical cancer progression. Mol. Biol. 37, 472–481.
Senchenko V.N., Liu J., Loginov W., Bazov I., Angeloni D., Seryogin Y., Ermilova V., Kazubskaya T., Garkavtseva R., Zabarovska V.I., Kashuba V.I., Kisselev L.L., Minna J.D., Lerman M.I., Klein G., Braga E.A., Zabarovsky E.R. 2004. Discovery of frequent homozygous deletions in chromosome 3p21.3 LUCA and AP20 regions in renal, lung, and breast carcinomas. Oncogene. 23, 5719–5728.
Kashuba V.I., Li J., Wang F., Senchenko V.N., Protopopov A., Malyukova A., Kutsenko A.S., Kadyrova E., Zabarovska V.I., Muravenko O.V., Zelenin A.V., Kisselev L.L., Kuzmin I., Minna J.D., Winberg G., Ernberg I., Braga E., Lerman M.I., Klein G., Zabarovsky E.R. 2004. RBSP3 (HYA22) is a tumor suppressor gene implicated in major epithelial malignancies. Proc. Natl. Acad. Sci. USA. 101, 4906–4911.
Fieder M., Muller-Holzner E., Viertler H.P., Widschwend ter A., Laich A., Pfister G., Spoden G.A., Jansen-Durr P., Zwerschke W. 2004. High level HPV-16 E7 oncoprotein expression correlates with reduced pRb-levels in cervical biopsies. FASEB J. 10, 1120–1122.
Tringler B., Gup C.J., Singh M., Groshong S., Shroyer A.L., Heinz D.E., Shroyer K.R. 2004. Evaluation of p16 and Rb expression in cervical squamous and grandular neoplasia. Human Pathol. 35, 689–696.
Helt A.M., Galloway D.A. 2003. Mechanisms by which DNA tumor virus oncoproteins target the Rb family of pocket proteins. Carcinogenesis. 24, 159–169.
Maniatis T., Fritsch E.F., Sambrook J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Lab. Press, pp. 186–196.
Boulay J.-L., Reuter J., Ristchard R., Terracciano L., Herrmann R., Rochlist C. 1999. Gene dosage quantitative real-time PCR. BioTechniques. 27, 228–232.
Livak K.J., Flood S.J., Marmaro J., Giusti W., Deets K. 1995. Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization. PCR Methods Appl. 4, 357–362.
Radonic A., Thulke S., Mackay I.M., Landt O., Siegert W., Nitsche A. 2004. Guideline to reference gene selection for quantitative real-time PCR. Biochem. Biophys. Res. Commun. 313, 856–862.
Huggett J., Dheda K., Bustin S., Zumla A. 2005. Real-time RT-PCR normalization: Strategies and considerations. Genes Immun. 6, 279–284.
Wong M.L., Medrano J.F. 2005. Real-time PCR for mRNA quantification. BioTechniques. 39, 1–11.
Kisselev L.L., Senchenko V.N., Oparina N.Yu., Braga E.A., Zabarovsky E.R. 2005. Tumor suppressor genes localized on the short arm of human chromosome 3. Mol. Meditsina. 3, 17–28.
Yeo M., Lin P.S., Dahmus M.E., Gill G.N. 2003. A novel RNA polymerase II C-terminal domain phosphatase that preferentially dephosphorylates serine 5. J. Biol. Chem. 11, 26,078–26,085.
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Original Russian Text © E.A. Anedchenko, N.P. Kisseljova, A.A. Dmitriev, F.L. Kisseljov, E.R. Zabarovsky, V.N. Senchenko, 2007, published in Molekulyarnaya Biologiya, 2007, Vol. 41, No. 1, pp. 86–95.
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Anedchenko, E.A., Kisseljova, N.P., Dmitriev, A.A. et al. Tumor suppressor gene RBSP3 in cervical carcinoma: Copy number and transcription level. Mol Biol 41, 77–85 (2007). https://doi.org/10.1134/S0026893307010116
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DOI: https://doi.org/10.1134/S0026893307010116