利托那韦对卡波氏肉瘤相关疱疹病毒感染细胞LYN表达的影响

张芳; 陈颖; 郭峰; 徐益明; 谭晓华; 杨磊; 李锋; 姬玉; 李冬妹

doi:10.3971/j.issn.1000-8578.2017.03.003

利托那韦对卡波氏肉瘤相关疱疹病毒感染细胞LYN表达的影响

Effects of Ritonavir on Expression of LYN in Kaposi's Sarcoma-associated Herpesvirus-infected Cells

摘要

摘要:
目的探讨蛋白酶抑制剂利托那韦对卡波氏肉瘤相关疱疹病毒（Kaposi’s sarcoma associated herpesvirus, KSHV）感染细胞LYN表达的影响。
方法原代培养人脐静脉内皮细胞HUVECs，从BCBL-1中提取KSHV病毒感染HUVECs，PCR扩增KS330₂₃₃证实感染成功。CCK8法测定半数抑制浓度，CCK8法检测细胞增殖能力，Western blot检测LYN表达及p-PI3K和p-AKT水平。
结果 RTV作用于细胞的IC₅₀为25 μmol/L。RTV与LYN抑制剂PP2能抑制KSHV感染的HUVECs的增殖和LYN、p-PI3K和p-AKT的表达（P < 0.05）。
结论 RTV可通过抑制LYN及其下游的PI3K/AKT信号通路抑制KSHV感染细胞HUVECs的增殖。

Abstract:
Objective To detect the effects of protein inhibitor ritonavir (RTV) on the expression of LYN in Kaposi' s sarcoma associated herpesvirus (KSHV) infected cells.
Methods Primary Human Umbilical Vein Endothelial Cells (HUVECs) were cultivated by enzyme infusion method. KSHV were extracted from BCBL-1 cells and infected HUVECs. PCR was used to amplify KS330₂₃₃ fragment to verify the KSHV infection. We used CCK8 method to detect the IC₅₀ of RTV on HUVECs and the proliferation ability of the cells. Western blot was used to detect LYN expression, p-PI3K and p-AKT levels.
Results RTV could inhibit the growth of KSHV-infected HUVECs in a concentration-dependent manner (P < 0.05). The IC₅₀ of RTV effect on cells was 25μmol/L. RTV could inhibit LYN, p-PI3K and p-AKT expression.
Conclusion RTV could inhibit the proliferation of KSHV-infected HUVECs through inhibiting LYN and PI3K/AKT signaling pathways.

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