Dissertation
Dissertation > Medicine, health > Oncology > Genitourinary tumors > Male genitalia tumors > Prostate cancer

The Eukaryotic Expression of HIV-1 Rev, Its Effect on the Lytic Cycle Replication of KSHV and the Relationship between KSHV and Prostate Cancer Cells

Author LiJiuMing
Tutor HuaLiXin
School Nanjing Medical University
Course Urology
Keywords Human immunodeficiency virus type 1 Virus particle protein regulatory factors Human herpes virus type 8 Copy Infection
CLC R737.25
Type Master's thesis
Year 2009
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Human immunodeficiency virus type 1 ( human as immunodeficiency or virus type 1 , of HIV-1 ) infection significantly with the increase of Kaposi's sarcoma -related herpes virus ( Kaposi's 's sarcoma cells in - associated herpesvirus , KSHV ) infection by occurrence of Kaposi economic sarcoma ( Kaposi's 's sarcoma cells in , KS ) of risk. KSHV infection is a necessary condition of KS occurred , but enough to trigger there is no when other cofactors role is KS, promote KSHV triggered KS factors , however , are not completely understood . Before our research has confirmed HIV-1 - trans- activating protein (transactivative transcription protein, Tat) is activated to KSHV lytic cycle replication factors . This paper plasmid containing the HIV-1 virus particle protein expression regulatory factor (regulator of virion protein expression, Rev) gene encoding the prev - Flag recombinant plasmids and restriction enzyme digestion and sequence determination ; prev- Flag recombinant plasmids were transiently transfected primary exudative lymphoma [primary effusion lymphoma, PEL, also known as body cavity lymphomas (body cavity-based lymphoma, BCBL)] sources BCBL-1 cells and mouse embryonic fibroblasts NIH/3T3 by RT - PCR , Western blot were detected from the mRNA and protein levels of the Rev gene expression ; BCBL-1 cell total RNA extract transiently transfected with the recombinant plasmid of pRev-Flag , Real-time PCR detection of KSHV minor capsid protein coding genes ORF26 KSHV cycles the copy switch gene ORF50 mRNA and RT-PCR detection the ORF26 mRNA transcription level ; BCBL - 1 cells and PC-3 culture change in cell morphology observed cultured PC-3 cells , RT-PCR was used to detect the expression of KSHV ORF26 . The nucleic acid sequence analysis showed that the cloned the Rev gene sequences in GenBank registered Rev sequence 100% homologous RT-PCR and Western blot analysis detected a specific band of in the Rev expected position . Real-time PCR and RT-PCR analysis showed that the Rev gene encodes a protein can reduce KSHV transcriptional level . The cells were cultured PC-3 cell shape change , RT-PCR detected ORF26 expression . This study successfully constructed a recombinant plasmid containing the sequence of Rev gene and obtain the correct expression in eukaryotic cells BCBL-1 and NIH/3T3 cells ; preliminary exploration showed that the the Rev gene encoding protein can inhibit KSHV lytic cycle replication ; KSHV can infect prostate cancer .

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