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

Human immunodeficiency virus type 1 (HIV-1) infection significantly increases the risk of Kaposi's sarcoma (KS) occurrence in individuals infected with Kaposi's sarcoma-associated herpesvirus (KSHV). KSHV infection appears to be necessary but not sufficient for KS development without other cofactors. However, factors that facilitate KSHV to cause KS have not been well defined. Previously, we determined that HIV-1 transactivative transcription protein (Tat) was one of the cofactors that activated lytic cycle replication of KSHV. Here, after identification with enzyme digestion and nucleotide sequences analysis, the recombinant plasmid of regulator of HIV-1 virion protein expression (Rev) (pRev-Flag) was transiently transfected into BCBL-1 cells and NIH/3T3 cells. The expression of pRev-Flag mRNA and protein in both cells was detected by RT-PCR and Western blot, respectively. Subsequently, RNA was obtained from BCBL-1 cells transfected with pRev-Flag. Real-time PCR and RT-PCR was carried out to evaluate the expression of ORF26 (encoding the minor capsid protein)/ORF50 (switch gene of KSHV cycle replication) mRNA of KSHV. At last, we describe our attempts to infect PC-3 cells (prostate cancer cell lines) with KSHV by coculture with BCBL-1 cells (KSHV-positive cell lines). Nucleotide sequences analysis indicated that the cloned Rev sequence was 100% homology with Rev gene previously registered in GenBank. The specific band was detected at the expected place by both RT-PCR and Western blot. In addition, the expression of ORF26/ORF50 mRNA in BCBL-1 cells transfected with pRev-Flag was weaker than the control. We detected the expression of ORF26 mRNA in coculture PC-3 cells. These findings suggest that Rev gene was correctly expressed in BCBL-1 cells and NIH/3T3 cells, and Rev gene might inhibit KSHV lytic cycle replication. KSHV might infect PC-3 cells.