Modified Shenlingbaizhu Decoction represses the pluripotency of colorectal cancer stem cells by inhibiting TGF-β mediated EMT program
Modified Shenlingbaizhu Decoction represses the pluripotency of colorectal cancer stem cells by inhibiting TGF-β mediated EMT program
复制标题
加味参苓白术汤通过抑制TGF-β介导的EMT程序抑制大肠癌干细胞的多能性
DOI：
10.1016/j.phymed.2022.154234
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发表时间：
2022-06-08
期刊：
影响因子：
7.9
通讯作者：
Sun, Xuegang
中科院分区：
文献类型：
--
作者：
Dai, Yu;Wang, Hao;Sun, Xuegang
背景：参灵白术加减汤（MSD）利用多种植物药物治疗结直肠癌（CRC）。结直肠癌干细胞（CSCs）已被证明与结直肠癌的进展和转移密切相关。默沙东对CSCs的抑制作用机制尚未确定。目的：了解MSD如何抑制CSCs的多能性并阻碍EMT程序。方法：采用高效液相色谱法对丹参提取物的成分进行表征。将BALB/c-nu小鼠移植到EGFP标记的SW480 CRC细胞中，记录不同剂量MSD治疗前后肿瘤的重量和体积。用酶联免疫吸附法测定tgf - β 1的浓度。为了阐明EMT与MSD调控的CSCs之间的逻辑关系，我们在荷瘤小鼠和多种CRC细胞系中采用了tgf - β /Smad抑制剂和激活剂。用流式细胞术分析肿瘤干细胞标志物。采用CCK-8、伤口愈合和侵袭实验进行体外细胞运动和活力分析。免疫组化（IHC）和western blotting （WB）检测蛋白表达。采集结果用GraphPad Prism 8.0进行统计分析。结果：MSD治疗可显著减小小鼠结直肠癌肿瘤的大小，降低血清tgf - β 1的含量。重要的是，MSD显著降低了肿瘤组织中多能因子的表达，抑制了CD133+干细胞的表达。tgf - β /Smad抑制剂中和EMT信号并通过SMAD2/3的去磷酸化降低多能性。同样，MSD通过限制体内tgf - β /Smad信号诱导的EMT来减弱多能性。默沙东抑制结直肠癌细胞的增殖、迁移和侵袭。结论：默沙东具有抑制结直肠癌生长的作用。它通过抑制tgf -8诱导的EMT程序来抑制csc的多能性。
Background: The Modified Shenlingbaizhu Decoction (MSD) utilizes various phytomedicines has been applied to treat colorectal cancer (CRC). Colorectal cancer stem cells (CSCs) have proven to be tightly associated with CRC progression and metastasis. The mechanism of MSD's inhibitory effect on CSCs has not been determined.Purpose: To figure out how MSD inhibits the pluripotency of CSCs and impedes the EMT program.Methods: The ingredients of MSD extracts were characterized by high-performance liquid chromatography (HPLC). BALB/c-nu mice were transplanted into EGFP labeled SW480 CRC cells and the tumor weight and volume were recorded before and after various doses of MSD treatment. The concentration of TGF-beta 1 was quantified with an Enzyme-linked immunosorbent assay. To delineate the logical relationship between EMT and CSCs regulated by MSD, TGF-beta/Smad inhibitor and activator were adopted in tumor-bearing mice and diverse CRC cell lines. Cancer stem cell markers were analyzed by flow cytometry. In vitro analysis of cell motility and viability were done using CCK-8, wound healing, and invasion assay. Immunohistochemistry (IHC) and western blotting (WB) were used for detecting protein expression. The collected results were statistically analyzed with GraphPad Prism 8.0.Results: MSD treatment significantly reduced the size of colorectal cancer tumors and lowered the serum content of TGF-beta 1 in mice. Importantly, MSD markedly reduced the expression of pluripotent factors and depressed CD133+ stem cells in the tumor tissues. The TGF-beta/Smad inhibitor neutralized the EMT signaling and lowered the pluripotency by dephosphorylation of SMAD2/3. Similarly, MSD attenuated the pluripotency by limiting TGF-beta/Smad signaling-induced EMT in vivo. MSD inhibited colorectal cancer cell proliferation, migration, and invasion.Conclusions: MSD inhibits the growth of colorectal cancer. It dampens the pluripotency of CSCs by repressing the TGF-8-induced EMT program.