Establishment and its significance of the method for the detection of HPV16 early gene E6-related transcripts in cervical cancer
1.Department of Microbiology and Immunology, Institute of Molecular Virology and Immunology, Wenzhou Medical University, Wenzhou, 325035; 2.Department of Obstetrics and Gynecology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000; 3.School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, 325035
LU Xulian1,ZOU Ruanmin2,CHEN Xiangmin3, et al. Establishment and its significance of the method for the detection of HPV16 early gene E6-related transcripts in cervical cancer[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2014, 44(5): 324-.
Abstract：Objective: To establish the method for the detection of HPV16 early gene E6-related transcripts and analyze the HPV16 E6-associated transcription during the development of cervical cancer. Methods: Sixty- three HPV16 positive cervical tissues, including 8 LSIL (low-grade squamous intraepithelial lesion, 38 HSIL (high-grade squamous intraepithelial lesion) and 17 cervical cancer, were collected. The extracted RNAs were then reversed by a RT primer containing a conserved sequences at 5’ end. The cDNAs were then used as the templates to amplify the transcripts using the HPV16 E6 specific primers (P1) and the conserved sequence (P0). After separated by 2.5% agarose gel electrophoresis, the amplification products were detected by Southern blotting using an E6-specific probe to verify the HPV16 E6-related transcripts. Results: The method for the detection of HPV16 E6-related transcripts was established. Further analysis showed that HPV16 oncogenes transcription patterns were significantly different in the tissues of LSIL, HSIL, and cervical cancer. The types of HPV16 E6-associated transcripts increased during the development from LSIL to cervical cancer. Conclusion: The method for the detection of HPV16 E6-related transcripts is successfully established and the patterns of HPV16 E6-associated transcripts are closely associated with the malignant degree of cervical neoplasm and may be involved in the development of cervical cancer.
