A non-radioactive PCR-SSCP analysis allows to distinguish between HPV 16 European and Asian-American variants in squamous cell carcinomas of the uterine cervix in Colombia.
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.


CONCLUSIONS
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.
Grupo de Investigación en Biología del Cáncer, Instituto Nacional de Cancerología, Calle 1a. No. 9-85, Bogota, D C, Colombia. dajup63@yahoo.com
Pablo
P
Moreno-Acosta
Mónica
M
Molano
Antonio
A
Huertas
Myriam Sánchez
MS
de Gómez
Alfredo
A
Romero
Mauricio
M
González
María
M
Mercedes Bravo
Alejandro
A
García-Carrancá
D000328:Adult; D000368:Aged; D001213:Asian / Q000706:statistics & numerical data; D002294:Carcinoma, Squamous Cell / Q000453:epidemiology / Q000208:ethnology / Q000821:virology*; D003105:Colombia / Q000453:epidemiology; D004742:Enhancer Elements, Genetic; D005260:Female; D052162:Human papillomavirus 16 / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008875:Middle Aged; D009856:Oncogene Proteins, Viral / Q000235:genetics; D030361:Papillomavirus Infections / Q000453:epidemiology / Q000208:ethnology / Q000821:virology*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012097:Repressor Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000453:epidemiology / Q000208:ethnology / Q000821:virology*; D044465:White People / Q000706:statistics & numerical data
C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins