Increased expression of 70 kD heat shock protein in cultured primary human keratinocytes induced by human papillomavirus 16 E6/E7 gene
- Author:
- No author available
- Journal Title:
- CHINESE MEDICAL JOURNAL
- Issue:
- 24
- DOI:
- No doi available
- Key Word:
- heat shock protein 70;keratinocytes;human papillomavirus 16 E6/E7
Abstract： Background Heat shock protein 70 (HSP70) is expressed highly in epithelial tumours associated closely with human papillomavirus 16 (HPV16) infections. However, evidence about the direct relationship between HSP70 expression and HPVs infections are still lacking. In the present study, we examined the expression of HSP70 in keratinocytes introduced with HPV16 E6/E7 oncogenes. Methods Stable transfected cells were established by transfection of the plasmids pLXSN16E6/E7 into cultured primary keratinocytes and subsequently selected by plasmid specific selection antibiotic (G418) at the required concentration. The expression of HSP70 in pLXSN16E6/E7 transfected keratinocytes was determined by Western blot. The correlation of HSP70 expression and E6/E7 transfection was further confirmed by doubly labelled immunofluorescent staining.Results Compared to non-transfected keratinocytes, there was a significant trend for higher levels of HSP70 in pLXSN16E6/E7 transfected keratinocytes. Doubly labelled immunofluorescent staining experiment showed that the co-localization of HPV16 E6/E7 and HSP70 in transfected keratinocytes was observed and increased expression of HSP70 was strongly associated with the transfection of HPV16 E6/E7.Conclusions Our studies demonstrated increased levels of HSP70 proteins in keratinocytes stably transfected by HPV16 E6/E7 oncogenes. It suggests that the expression of HSP70 is modulated by HPV16 E6/E7 proteins, which may be involved in HPV16 E6/E7 induced immortalization.
- [1]E J, Androphy.Molecular biology of human papillomavirus infection and oncogenesis.[J].1994,103(2).
- [2]X, Liu,L, Ye,J, Wang,etc.Expression of heat shock protein 90 beta in human gastric cancer tissue and SGC7901/VCR of MDR-type gastric cancer cell line.[J].1999,112.
- [3]Shimada H,Ochiai T,Kobayashi S,etc.Immortalization of human esophageal keratinocytes by E6 and E7 of human papillomavirus type 16.[J].2001,19(1).
- [4]Frauke, Fehrmann,Laimonis A, Laimins.Human papillomaviruses: targeting differentiating epithelial cells for malignant transformation.[J].2003,22(33).
- [5]李清泉,胡克,潘显光,等.Detection of Human Papillomavirus Types 16, 18 Dna Related Sequences in Bronchogenic Carcinoma by Polymerase Chain Reaction[J].1995,(8).
- [6]M, J??ttel?.Over-expression of hsp70 confers tumorigenicity to mouse fibrosarcoma cells.[J].1995,60.
- [7]X F, Pei.The human papillomavirus E6/E7 genes induce discordant changes in the expression of cell growth regulatory proteins.[J].1996,17.
- [8]Tsokos GC,Kiang JG.Heat shock protein 70 kDa: molecular biology, biochemistry, and physiology.[J].1998,80(2).
- [9]K K, Kim,T J, Jang,J R, Kim.HSP70 and ER expression in cervical intraepithelial neoplasia and cervical cancer.[J].1998,13.
- [10]C A, Finlay,P W, Hinds,T H, Tan,etc.Activating mutations for transformation by p53 produce a gene product that forms an hsc70-p53 complex with an altered half-life.[J].1988,8.
- [11]J, Kr?ll.Molecular chaperones and the process of cellular immortalization in vitro.[J].2002,3(3).
- [12]Kensyu Kawanishi M.D.,Hitoshi Shiozaki M.D.,Yuichiro Doki M.D.,etc.Prognostic significance of heat shock proteins 27 and 70 in patients with squamous cell carcinoma of the esophagus[J].1999,85(8).
- [13]M, Ferrarini,S, Heltai,M R, Zocchi,etc.Unusual expression and localization of heat-shock proteins in human tumor cells.[J].1992,51.
- [14]Tewari D,Hagmar B,Barrera Saldana HA,etc.Conserved methylation patterns of human papillomavirus type 16 DNA in asymptomatic infection and cervical neoplasia.[J].2004,78(23).
- [15].Specific interaction between the p53 cellular tumour antigen and major heat shock proteins[J].1986,320(6058).
- [16]Matsunami N,Meloni Ehrig-A,Jorgensen CM,etc.Persistent transgene expression and normal differentiation of immortalized human keratinocytes in vivo.[J].1999,112(2).
- [17]B, Phillips,K, Abravaya,R I, Morimoto.Analysis of the specificity and mechanism of transcriptional activation of the human hsp70 gene during infection by DNA viruses.[J].1991,65.
- [18]Quentin T,Dressel R,Walter L,etc.Heat shock protein 70 is able to prevent heat shock-induced resistance of target cells to CTL.[J].2000,164(5).