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肿瘤相关巨噬细胞对肾透明细胞癌增殖作用及其分子机制的影响 |
Sunitinib inhibits PDPK1-mediated PKG1 phosphorylation and inhibits proliferation of renal clear cell carcinoma by inhibiting macrophage secretion of IL-6. |
投稿时间:2019-03-20 修订日期:2019-03-20 |
DOI: |
中文关键词: 巨噬细胞 透明细胞癌 IL-6 PDPK1 PKG1 |
英文关键词: |
基金项目:陕西省自然科学基金(2017SZ3428JGF0) |
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中文摘要: |
目的 探讨肿瘤相关巨噬细胞(Tumor Associated Macrophages, TAMs)分泌IL-6从而促进PDPK1介导的PKG1磷酸化,在此基础上研究舒尼替尼是否通过间接作用肿瘤相关巨噬细胞从而抑制肾透明细胞癌的增殖及其分子机制。方法 将单核细胞系(THP-1)细胞诱导为 TAMs,同时采用细胞共培养方法将TAMs和肾透明细胞癌ACHN共培养,CCK-8和克隆形成实验检测共培养下舒尼替尼对肾透明细胞癌ACHN细胞增殖能力的影响;采用皮下接种方法建立裸鼠皮下移植瘤模型,观察舒尼替尼对裸鼠皮下成瘤能力及肿瘤体积大小的影响;ELISA检测舒尼替尼对巨噬细胞分泌TNF-α、IL-10和IL-6等肿瘤诱导因子的作用。Western Blot检测舒尼替尼对磷酸肌醇依赖性蛋白激酶1 (Phosphoinositol Dependent Protein Kinase 1,PDPK1)介导磷酸甘油酸激酶1(Phosphoglycerate Kinase 1, PKG1)的磷酸化作用的影响。结果 显微镜观察结果显示肿瘤相关巨噬细胞促进了ACHN细胞增殖作,而舒尼替尼可以抑制其促增殖作用;CCK-8和克隆形成实验表明TAMs促进了ACHN克隆形成能力,但是其克隆形成能力可以被舒尼替尼抑制;ELISA实验结果表明舒尼替尼可以抑制巨噬细胞分泌TGF-β、IL-10和IL-6等肿瘤诱导因子的表达;动物实验证明舒尼替尼可以抑制裸鼠皮下成瘤能力和移植瘤的成长;舒尼替尼通过抑制巨噬细胞分泌IL-6的作用阻断PDPK1介导PKG1的磷酸化;结论 舒尼替尼通过抑制巨噬细胞分泌IL-6的作用抑制了PDPK1介导PKG1的磷酸化作用,从而抑制了肾透明细胞癌的增殖作用。 |
英文摘要: |
Objective To investigate the secretion of IL-6 by Tumor-associated macrophages (TAMs) to promote PDPK1-mediated phosphorylation of PKG1. Based on this, we investigated whether sunitinib can indirectly affect tumor-associated macrophages.Thereby inhibiting the proliferation of renal clear cell carcinoma and its molecular mechanism. Methods Mononuclear cell line (THP-1) cells were induced into TAMs, and TAMs and renal clear cell carcinoma ACHN were co-cultured by cell co-culture. CCK-8 and clone formation assays were used to detect sunitinib on kidney. Effect of proliferation of clear cell carcinoma ACHN cells; subcutaneous vaccination was used to establish a subcutaneous xenograft model in nude mice, and the effect of sunitinib on subcutaneous tumorigenicity and tumor volume in nude mice was observed. ELISA detects the effect of sunitinib on the secretion of tumor-inducing factors such as TGF-β, IL-10 and IL-6 by macrophages. Western Blot was used to examine the effect of sunitinib on PDPK1-mediated phosphorylation of PKG1. Results Microscopic observation showed that tumor-associated macrophages promoted the proliferation of ACHN cells, while sunitinib could inhibit its proliferation. CCK-8 and colony formation experiments indicated that TAMs promoted ACHN clonality, but their ability to clone was inhibited by sunitinib. The results of ELISA showed that sunitinib can inhibit the expression of tumor-inducing factors such as TNF-α, TGF-β and IL-6 secreted by macrophages. Animal experiments demonstrate that sunitinib can inhibit the tumor formation ability and xenograft of nude mice. growing up.Sunitinib blocks PDPK1-mediated phosphorylation of PKG1 by inhibiting the secretion of IL-6 by macrophages. Conclusion Sunitinib inhibits PDPK1-mediated phosphorylation of PKG1 by inhibiting the secretion of IL-6 by macrophages, thereby inhibiting the proliferation of renal clear cell carcinoma. |
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