Moreno-Acosta Pablo, Molano Mónica, Huertas Antonio, de Gómez Myriam Sánchez, Romero Alfredo, González Mauricio, Mercedes Bravo María, García-Carrancá Alejandro
Grupo de Investigación en Biología del Cáncer, Instituto Nacional de Cancerología, Calle 1a. No. 9-85, Bogota, D C, Colombia.
Virus Genes. 2008 Aug;37(1):22-30. doi: 10.1007/s11262-008-0239-8. Epub 2008 May 30.
Human Papillomavirus type 16 (HPV 16) DNA is regularly found in around 50% of all cervical carcinomas. Variants of this type have been found associated with different risks for cervical cancer development. Presence of HPV 16 variants in Colombia has not been previously reported. The aims of this study were to assess the feasibility of non-radioactive PCR-SSCP (polymerase chain reaction single-strand conformation polymorphism) analysis for determination of variability of ORF of E6, variability in the enhancer sequence of the LCR, and for establishment of the distribution of HPV 16 variants in invasive squamous cell carcinoma of the uterine cervix in Colombian women. Biopsies from 59 patients at the Instituto Nacional de Cancerología (INC) in Bogotá (Colombia) were collected. HPV detection was performed using universal primers. HPV 16 variants were detected by non-radioactive single-stranded conformational polymorphism (SSCP) analysis and direct sequencing. HPV 16 was detected in 57.6% of the tumors. The European branch was identified in 88.2% of the samples with the E-G350 class being the most prevalent variant (41.1%). The Asian-American branch was identified in 8.8% of the samples. Within this group it was possible to distinguish between c and a classes. It was not possible to determine the branch in 2.9% of the cases. A nucleotide transition (G to A) at position 7521 was the most prevalent variation (80%) found in the enhancer sequence of the LCR region.
A non-radioactive PCR-SSCP analysis allowed us to distinguish between European and Asian-American branches of HPV 16, and to distinguish among classes in squamous cell carcinomas of the uterine cervix in Colombia. This method is an excellent alternative that can be used as a screening tool for identification of HPV 16 variants.
在所有宫颈癌中,约50% 经常可检测到16型人乳头瘤病毒(HPV 16)DNA。已发现该类型的变体与宫颈癌发生的不同风险相关。此前尚未报道过哥伦比亚存在HPV 16变体。本研究的目的是评估非放射性聚合酶链反应 - 单链构象多态性(PCR - SSCP)分析用于确定E6开放阅读框变异性、长控制区(LCR)增强子序列变异性以及确定哥伦比亚女性子宫颈浸润性鳞状细胞癌中HPV 16变体分布的可行性。收集了来自哥伦比亚波哥大国家癌症研究所(INC)的59例患者的活检样本。使用通用引物进行HPV检测。通过非放射性单链构象多态性(SSCP)分析和直接测序检测HPV 16变体。在57.6%的肿瘤中检测到HPV 16。在88.2%的样本中鉴定出欧洲分支,其中E - G350类别是最常见的变体(41.1%)。在8.8%的样本中鉴定出亚美分支。在该组中可以区分c类和a类。在2.9%的病例中无法确定分支。在LCR区域增强子序列中发现的最常见变异(80%)是第7521位核苷酸的转换(G到A)。
非放射性PCR - SSCP分析使我们能够区分HPV 16的欧洲和亚美分支,并区分哥伦比亚子宫颈鳞状细胞癌中的不同类别。该方法是一种优秀的替代方法,可作为鉴定HPV 16变体的筛查工具。