Kuglik Petr, Smetana Jan, Vallova Vladimira, Moukova Lucie, Kasikova Katerina, Cvanova Michaela, Brozova Lucie
Laboratory of Molecular Cytogenetics, Institute of Experimental Biology, Faculty of Science, Masaryk University Brno, Czech Republic ; Department of Medical Genetics, University Hospital Brno, Czech Republic.
Department of Gynecological Oncology, Masaryk Memorial Cancer Institute Brno, Czech Republic.
Int J Clin Exp Pathol. 2014 Jul 15;7(8):5071-82. eCollection 2014.
Alterations in the genome that lead to changes in DNA sequence copy number are characteristic features of solid tumors. We used CGH+SNP microarray and HPV-FISH techniques for detailed screening of copy number alterations (CNAs) in a cohort of 26 patients with cervical carcinoma (CC). This approach identified CNAs in 96.2% (25/26) of tumors. Array-CGH discovered CNAs in 73.1% (19/26) of samples, HPV-FISH experiments revealed CNAs in additional 23.1% (6/26) of samples. Common gains of genetic sequences were observed in 3q (50.0%), 1q (42.4%), 19q (23.1%), while losses were frequently found in 11q (30.8%), 4q (23.1%) and 13q (19.2%). Chromosomal regions involved in loss of heterozygosity were observed in 15.4% of samples in 8q21, 11q23, 14q21 and 18q12.2. Incidence of gain 3q was associated with HPV 16 and HPV 18 positive samples and simultaneous presence of gain 1q (P = 0.033). We did not found a correlation between incidence of CNAs identified by array-CGH and HPV strain infection and incidence of lymph node metastases. Subsequently, HPV-FISH was used for validation of array-CGH results in 23 patients for incidence of hTERC (3q26) and MYC (8q24) amplification. Using HPV-FISH, we found chromosomal lesions of hTERC in 87.0% and MYC in 65.2% of specimens. Our findings confirmed the important role of HPV infection and specific genomic alterations in the development of invasive cervical cancer. This study also indicates that CGH+SNP microarrays allow detecting genome-wide CNAs and copy-neutral loss of heterozygosity more precisely, however, it may be less sensitive than FISH in samples with low level clonal CNAs.
基因组中导致DNA序列拷贝数改变的变化是实体瘤的特征性表现。我们使用比较基因组杂交联合单核苷酸多态性(CGH+SNP)微阵列和人乳头瘤病毒荧光原位杂交(HPV-FISH)技术,对26例宫颈癌(CC)患者的队列进行拷贝数改变(CNA)的详细筛查。该方法在96.2%(25/26)的肿瘤中检测到CNA。阵列比较基因组杂交(Array-CGH)在73.1%(19/26)的样本中发现了CNA,HPV-FISH实验在另外23.1%(6/26)的样本中发现了CNA。常见的基因序列增益出现在3q(50.0%)、1q(42.4%)、19q(23.1%),而缺失则常见于11q(30.8%)、4q(23.1%)和13q(19.2%)。在8q21、11q23、14q21和18q12.2的15.4%的样本中观察到杂合性缺失所涉及的染色体区域。3q增益的发生率与HPV 16和HPV 18阳性样本以及1q增益的同时存在相关(P = 0.033)。我们未发现Array-CGH鉴定的CNA发生率与HPV毒株感染及淋巴结转移发生率之间存在相关性。随后,使用HPV-FISH对23例患者中hTERC(3q26)和MYC(8q24)扩增的发生率进行验证,以确认Array-CGH结果。使用HPV-FISH,我们在87.0%的标本中发现了hTERC的染色体病变,在65.2%的标本中发现了MYC的染色体病变。我们的研究结果证实了HPV感染和特定基因组改变在浸润性宫颈癌发生发展中的重要作用。本研究还表明,CGH+SNP微阵列能够更精确地检测全基因组CNA和拷贝数中性杂合性缺失,然而,在低水平克隆CNA的样本中,其敏感性可能低于FISH。