目的 从抑制信号转导和转录激活因子3(STAT3)磷酸化(p-STAT3)改变肿瘤微环境(tumor microenvironment, TME)角度探讨隐丹参酮抗弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)效应及其可能的分子机制。方法 将SUDHL-4细胞株传代培养后接种NOD-SCID小鼠建立DLBCL小鼠模型,分为模型组、隐丹参酮低剂量组和高剂量组,每周两次测量瘤体体积,干预17 d后处死小鼠取移植瘤,称量小鼠体质量和瘤体质量;ELISA检测外周血炎症因子及趋化因子配体2表达;Western blot检测移植瘤组织中STAT3蛋白及其磷酸化水平;免疫组化观察血管内皮生长因子(vascular endothelial growth factor, VEGF)表达情况;实时荧光定量PCR检测上皮间质转化(epithelial-mesenchymal transitions, EMT)相关指标(E-cadherin、MMP9和Vimentin)的mRNA水平。结果 低、高剂量组移植瘤体积及质量明显小于模型组,且高剂量组效果更为显著(P<0.001、P<0.01或P<0.05);IL-6、IL-10、TGF-β和CCL2在低、高剂量组中均较模型组小鼠表达显著降低,而IL-12呈现相反趋势,且高剂量组较低剂量组更明显(P<0.001、P<0.01或P<0.05);低、高剂量组小鼠移植瘤中p-STAT3和血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白及大部分EMT指标的mRNA表达显著低于模型组,且高剂量组抑制效果更好(P<0.001,P<0.01或P<0.05)。结论 隐丹参酮可下调STAT3磷酸化,改变肿瘤相关炎症因子、趋化因子CCL2,继而改变TME,抑制VEGF和EMT相关指标表达,最终抑制DLBCL小鼠移植瘤的生长。
Abstract
OBJECTIVE To investigate the effect of cryptotanshinone on tumor microenvironment(TME) and its possible molecular mechanism in mice with diffuse large B cell lymphoma(DLBCL) based on STAT3 phosphorylation. METHODS The model of xenograft tumor was established by SUDHL-4 cell line(DLBCL) in NOD-SCID mice. The model mice were divided into model group, cryptotanshinone low-dose group and high-dose group. The tumor volumes were measured twice a week, and the mice were killed after 17 days, the weights of the mice and the tumor were measured, the expression of inflammatory factors and CCL2 in peripheral blood were detected by Elisa, the levels of STAT3 protein and its phosphorylation in the xenograft tumor were detected by Western blot, the expression of VEGF was observed by immunohistochemistry, and the mRNA levels of E-cadherin, MMP9 and Vimentin were detected by real-time PCR. RESULTS The results showed that the volume and weight of the xenograft tumors in the low and high-dose groups were significantly lower than model group, while the effect was more pronounced in the high-dose group(P<0.001,P<0.01 or P<0.05), and the expressions of IL-6, IL-10, TGF-β and CCL2 in the low and high-dose groups were significantly lower than model group, while IL-12 showed the opposite trend, also the high-dose group was more pronounced(P<0.001, P<0.01 or P<0.05). The expressions of p-STAT3, VEGF and the most of EMT markers in the low and high-dose group were significantly lower than model group, and the high-dose group still has the best effect(P<0.001, P<0.01 or P<0.05). CONCLUSION Cryptotanshinone can inhibit the growth of xenograft tumor in DLBCL mice, which may be related to the changes of TME, including suppression of STAT3 phosphorylation and subsequent down-regulation of tumor-associated inflammatory cytokines, chemokine CCL2, VEGF expression, and EMT markers.
关键词
弥漫大B细胞淋巴瘤 /
STAT3磷酸化 /
隐丹参酮 /
肿瘤微环境 /
趋化因子配体2 /
上皮间质转化
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Key words
DLBCL /
STAT3 phosphorylation /
cryptotanshinone /
TME /
CCL2 /
EMT
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中图分类号：
R965
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参考文献
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脚注
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基金
浙江中医药大学自然科学青年探索项目资助(2021JKZKTS002A)
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