LukS-PV inhibits the proliferation of hepatocellular carcinoma cells by maintaining FOXO3 stability via the PI3K/AKT signaling pathway.
LukS-PV inhibits the proliferation of hepatocellular carcinoma cells by maintaining FOXO3 stability via the PI3K/AKT signaling pathway.
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DOI：
10.1016/j.cellsig.2022.110357
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发表时间：
2022-05
影响因子：
4.8
通讯作者：
Zheng-chao Nie;Lan Shi;K. Song;Xuexue Xu;Pengsheng Ding;B. Lu;Gang Wu;Xiaoling Ma
中科院分区：
文献类型：
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作者：
Zheng-chao Nie;Lan Shi;K. Song;Xuexue Xu;Pengsheng Ding;B. Lu;Gang Wu;Xiaoling Ma
肝细胞癌（Hepatocellular carcinoma，HCC）是常见的恶性肿瘤之一。LukS-PV是金黄色葡萄球菌（Staphylococcus aureus）分泌的Panton-Valvinylleukocidin（PVL）的S组分。叉头盒O3（FOXO 3）是转录因子FOXO亚家族的成员，其充当肿瘤抑制因子。本研究旨在探讨LukS-PV在肝癌细胞增殖中的作用及其可能的机制。我们用不同浓度的LukS-PV处理HCC细胞，并使用细胞计数试剂盒-8和集落形成测定来评估LukS-PV对细胞活力的影响。实时PCR和蛋白质印迹法分别用于分析mRNA和蛋白质表达水平。进行免疫荧光染色以检查FOXO 3的细胞内定位。免疫组化法检测FOXO 3及其下游靶基因的表达。蛋白质合成抑制剂放线菌酮和蛋白酶体抑制剂MG 132被用来探索LukS-PV调节FOXO 3的潜在机制。我们证明LukS-PV以浓度依赖性方式抑制肝癌细胞的增殖。LukS-PV在体外和体内均可上调FOXO 3的表达。此外，LukS-PV促进FOXO 3进入细胞核，随后调节下游靶基因的转录。此外，我们发现LukS-PV通过PI 3 K/AKT信号通路降低磷酸化FOXO 3的表达，并通过泛素-蛋白酶体途径维持FOXO 3蛋白的稳定性。综上所述，我们的数据表明，LukS-PV通过FOXO 3发挥抗癌活性。LukS-PV可能是一种有希望的肝癌治疗候选药物。
Hepatocellular carcinoma(HCC) is one of the common malignant tumors. LukS-PV is the S component of Panton-Valetine leukocidin(PVL) secreted byStaphylococcus aureus. Forkhead box O3 (FOXO3) is a member of the FOXO subfamily of transcription factors that acts as a tumor suppressor. In this study, we investigated the role of LukS-PV on the proliferation of HCC and explored possible mechanisms. We treated HCC cells with various concentrations of LukS-PV and evaluated the effect of LukS-PV on cell viability using the cell counting kit-8 and colony formation assays. Real-time PCR and western blot assays were used to analyze mRNA and protein expression levels, respectively. Immunofluorescence staining was performed to examine the intracellular localization of FOXO3. The expression of FOXO3 and its downstream target genes were analyzed by immunohistochemical staining. The protein synthesis inhibitor cycloheximide and the proteosome inhibitor MG132 were used to explore the potential mechanisms by which LukS-PV regulated FOXO3. We demonstrated that LukS-PV inhibited the proliferation of HCC cells in a concentration dependent manner. LukS-PV upregulated FOXO3 expression bothin vitroandin vivo. Moreover, LukS-PV facilitated the entry of FOXO3 into the nucleus and, subsequently, regulated the transcription of downstream target genes. In addition, we discovered that LukS-PV decreased the expression of phosphorylated FOXO3 through the PI3K/AKT signaling pathway and maintained FOXO3 protein stabilityviathe ubiquitin-proteasome pathway. Taken together, our data indicated that LukS-PV exert anticancer activities through FOXO3. LukS-PV may be a promising candidate for HCC treatment.