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目的探讨Kaposi肉瘤相关疱疹病毒(kaposi’s sarcoma associated herpesvirus,KSHV)感染对v-yes-1Yamaguchi肉瘤病毒相关同源癌基因(v-yes-1 Yamaguchi sarcoma viral related oncogene homolog,LYN)表达的影响,及其在卡波氏肉瘤(kaposi’s sarcoma,KS)发病机制中的作用。方法采用酶灌注法原代培养脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),运用免疫细胞化学检测CD34和CD31以验证HUVECs。从BCBL-1中提取KSHV病毒感染HUVECs,显微镜下观察细胞形态并拍照,PCR扩增KS330_(233)证实感染成功。采用Western blot检测LYN表达及p-PI3K和p-AKT水平。结果HUVECs中CD 31胞膜阳性和CD 34胞浆阳性,说明酶灌注法取得的细胞为HUVECs。KSHV感染HUVECs后细胞变为长梭形,LYN表达增高,同时p-PI3K和p-AKT水平也增高。结论 KSHV感染HUVECs促进LYN表达,且可能通过促进p-PI3K和p-AKT水平增高而活化PI3K/AKT信号通路。
Objective To investigate the effect of Kaposi’s sarcoma associated herpesvirus (KSHV) infection on the expression of v-yes-1 Yamaguchi sarcoma viral related oncogene homolog (LYN) In the pathogenesis of Kaposi’s sarcoma (KS). Methods Human umbilical vein endothelial cells (HUVECs) were primarily cultured by enzyme perfusion. CD34 and CD31 were detected by immunocytochemistry to verify HUVECs. HUVECs were infected with KSHV virus from BCBL-1, the morphology of the cells was observed under a microscope and photographed. KS330_ (233) PCR was used to confirm the infection. The expression of LYN and the levels of p-PI3K and p-AKT were detected by Western blot. Results HUVECs were positive for CD 31 and cytoplasm positive for CD 34, indicating that HUVECs were obtained by enzyme perfusion. After HUVECs were infected with KSHV, the cells became long fusiform, LYN expression increased, and the levels of p-PI3K and p-AKT also increased. Conclusion KSHV infected HUVECs can promote the expression of LYN, and may activate PI3K / AKT signaling pathway by increasing the levels of p-PI3K and p-AKT.