Expression of the H19 Oncofetal Gene in Premalignant Lesions of Cervical Cancer: A Potential Targeting Approach for Development of Nonsurgical Treatment of High-Risk Lesions
Background. Recent data suggest a role for H19 gene in promoting cancer transformation and progression. Cervical cancer, progresses from high-grade lesions (CIN3). At present, it is unclear if CIN lesions express H19. Objectives. To determine H19 expression in patient samples of CIN3 as well as the ability of a construct in which the promoter from the H19 gene drives expression of the diphtheria toxin A chain (DTA) to inhibit cervical cancer cell growth in vitro. Methods. H19 transcript levels were evaluated on 10 biopsies of CIN3 using in situ hybridization. PCR was used to examine H19 expression in cervical cancer cell lines and in two samples from a patient with cervical carcinoma. Cell lines were transfected with H19-DTA to determine its impact on cell number. Results. H19 gene was expressed in the area of CIN3 in 9 out of 10 samples. RT-PCR indicated expression of H19 in cervical cancer samples and in one of the three cell lines examined. Transfection of all cell lines with H19-DTA vector resulted in inhibited cell growth. Conclusions. H19 is expressed in the majority of CIN3 samples. These results suggest that most CIN3 lesions could be targeted by H19-DTA. Further in vivo preclinical studies are thus warranted. 1. Introduction Carcinoma of the uterine cervix is the second most common cancer worldwide. Every year, cervical cancer is diagnosed in about 500,000 women globally and is responsible for more than 280,000 deaths. While most cases of cervical cancer are diagnosed in developing countries, the USA National Institute of Health estimated that in the USA during 2010, 12,200 new cases of cervical cancer would be diagnosed and over 4000 women would die from this disease. The primary events leading to cervical dysplasia and carcinogenesis are most commonly related to infection with the human papilloma virus (HPV). Infection with HPV is diagnosed in as many as 99% of the women diagnosed with squamous cervical carcinoma cancer of the uterus. HPV is a double-stranded DNA virus that affects the growth and differentiation of cervical epithelial cells by interaction between the envelope proteins E6, E7 and tumor suppressor genes such as P53 and the retinoblastoma gene (Rb). Infection of cervical epithelial cells with the HPV causes the formation of early premalignant lesions, referred to as cervical intraepithelial neoplasia 1 (CIN1). In most cases, the cellular immune system eliminates these cells, resulting in a return to a normal epithelium. However, in some cases, these low-grade lesions progress to high-grade lesions (CIN3) and frank cervical
References
[1]
K. Middleton, W. Peh, S. Southern et al., “Organization of human papillomavirus productive cycle during neoplastic progression provides a basis for selection of diagnostic markers,” Journal of Virology, vol. 77, no. 19, pp. 10186–10201, 2003.
[2]
N. P. Caraway, A. Khanna, M. Dawlett et al., “Gain of the 3q26 region in cervicovaginal liquid-based pap preparations is associated with squamous intraepithelial lesions and squamous cell carcinoma,” Gynecologic Oncology, vol. 110, no. 1, pp. 37–42, 2008.
[3]
M. A. Whiteside, E. M. Siegel, and E. R. Unger, “Human papillomavirus and molecular considerations for cancer risk,” Cancer, vol. 113, no. 10, pp. 2981–2994, 2008.
[4]
S. J. Kim, S. E. Park, C. Lee et al., “Alterations in promoter usage and expression levels of insulin-like growth factor-II and H19 genes in cervical carcinoma exhibiting biallelic expression of IGF-II,” Biochimica et Biophysica Acta, vol. 1586, no. 3, pp. 307–315, 2002.
[5]
S. Douc-Rasy, M. Barrois, S. Fogel et al., “High incidence of loss of heterozygosity and abnormal imprinting of H19 and IGF2 genes in invasive cervical carcinomas. Uncoupling of H19 and IGF2 expression and biallelic hypomethylation of H19,” Oncogene, vol. 12, no. 2, pp. 423–430, 1996.
[6]
V. A. Erdmann, M. Z. Barciszewska, M. Szymanski, A. Hochberg, N. de Groot, and J. Barciszewski, “The non-coding RNAs as riboregulators,” Nucleic Acids Research, vol. 29, no. 1, pp. 189–193, 2001.
[7]
I. Ariel, H.-Q. Miao, X.-R. Ji et al., “Imprinted H19 oncofetal RNA is a candidate tumour marker for hepatocellular carcinoma,” Journal of Clinical Pathology—Molecular Pathology, vol. 51, no. 1, pp. 21–25, 1998.
[8]
I. Ariel, M. Sughayer, Y. Fellig et al., “The imprinted H19 gene is a marker of early recurrence in human bladder carcinoma,” Journal of Clinical Pathology—Molecular Pathology, vol. 53, no. 6, pp. 320–323, 2000.
[9]
O. Lustig-Yariv, E. Schulze, D. Komitowski et al., “The expression of the imprinted genes H19 and IGF-2 in choriocarcinoma cell lines. Is H19 a tumor suppressor gene?” Oncogene, vol. 15, no. 2, pp. 169–177, 1997.
[10]
H. Cui, P. Onyango, S. Brandenburg, Y. Wu, C.-L. Hsieh, and A. P. Feinberg, “Loss of imprinting in colorectal cancer linked to hypomethylation of H19 and IGF2,” Cancer Research, vol. 62, no. 22, pp. 6442–6446, 2002.
[11]
I. Ariel, O. Lustig, T. Schneider et al., “The imprinted H19 gene as a tumor marker in bladder carcinoma,” Urology, vol. 45, no. 2, pp. 335–338, 1995.
[12]
S. Ayesh, I. Matouk, T. Schneider et al., “Possible physiological role of H19 RNA,” Molecular Carcinogenesis, vol. 35, no. 2, pp. 63–74, 2002.
[13]
I. J. Matouk, N. DeGroot, S. Mezan et al., “The H19 non-coding RNA is essential for human tumor growth,” Plos One, vol. 2, no. 9, article e845, 2007.
[14]
P. A. Leighton, R. S. Ingram, J. Eggenschwiler, A. Efstratiadis, and S. M. Tilghman, “Disruption of imprinting caused by deletion of the H19 gene region in mice,” Nature, vol. 375, no. 6526, pp. 34–39, 1995.
[15]
M. L. Breitman, S. Clapoff, J. Rossant et al., “Genetic ablation: targeted expression of a toxin gene causes microphthalmia in transgenic mice,” Science, vol. 238, no. 4833, pp. 1563–1565, 1987.
[16]
I. Pastan, V. Chaudhary, and D. J. Fitzgerald, “Recombinant toxins as novel therapeutic agents,” Annual Review of Biochemistry, vol. 61, pp. 331–354, 1992.
[17]
P. Ohana, P. Schachter, B. Ayesh et al., “Regulatory sequences of H19 and IGF2 genes in DNA-based therapy of colorectal rat liver metastases,” Journal of Gene Medicine, vol. 7, no. 3, pp. 366–374, 2005.
[18]
A. Mizrahi, A. Czerniak, T. Levy et al., “Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences,” Journal of Translational Medicine, vol. 7, article 69, 2009.
[19]
P. Ohana, O. Bibi, I. Matouk et al., “Use of H19 regulatory sequences for targeted gene therapy in cancer,” International Journal of Cancer, vol. 98, no. 5, pp. 645–650, 2002.
[20]
P. Ohana, O. Gofrit, S. Ayesh et al., “Regulatory sequences of the H19 gene in DNA based therapy of bladder cancer,” Gene Therapy Molecular Biology, vol. 8, pp. 181–191, 2004.
[21]
L. Milligan, E. Antoine, C. Bisbal et al., “H19 gene expression is up-regulated exclusively by stabilization of the RNA during muscle cell differentiation,” Oncogene, vol. 19, no. 50, pp. 5810–5816, 2000.
[22]
B. Ayesh, I. Matouk, P. Ohana et al., “Inhibition of tumor growth by DT-A expressed under the control of IGF2 P3 and P4 promoter sequences,” Molecular Therapy, vol. 7, no. 4, pp. 535–541, 2003.
[23]
A. A. Sidi, P. Ohana, S. Benjamin et al., “Phase I/II marker lesion study of intravesical BC-819 DNA plasmid in H19 over expressing superficial bladder cancer refractory to bacillus calmette-guerin,” Journal of Urology, vol. 180, no. 6, pp. 2379–2383, 2008.
