The Key Laboratory of Molecular Biology, State Administration of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, Guangdong, China.
The Key Laboratory of Molecular Biology, State Administration of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, Guangdong, China; Shunde Hospital, Guangzhou University of Chinese Medicine, Foshan, 528333, Guangdong, China.
Phytomedicine. 2022 Aug;103:154234. doi: 10.1016/j.phymed.2022.154234. Epub 2022 Jun 1.
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.
To figure out how MSD inhibits the pluripotency of CSCs and impedes the EMT program.
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-β1 was quantified with an Enzyme-linked immunosorbent assay. To delineate the logical relationship between EMT and CSCs regulated by MSD, TGF-β/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.
MSD treatment significantly reduced the size of colorectal cancer tumors and lowered the serum content of TGF-β1 in mice. Importantly, MSD markedly reduced the expression of pluripotent factors and depressed CD133 stem cells in the tumor tissues. The TGF-β/Smad inhibitor neutralized the EMT signaling and lowered the pluripotency by dephosphorylation of SMAD2/3. Similarly, MSD attenuated the pluripotency by limiting TGF-β/Smad signaling-induced EMT in vivo. MSD inhibited colorectal cancer cell proliferation, migration, and invasion.
MSD inhibits the growth of colorectal cancer. It dampens the pluripotency of CSCs by repressing the TGF-β-induced EMT program.
参灵白术汤（MSD）采用多种植物药治疗结直肠癌（CRC）。结直肠肿瘤干细胞（CSC）已被证明与 CRC 的进展和转移密切相关。MSD 抑制 CSC 的作用机制尚未确定。
探讨 MSD 如何抑制 CSC 的多能性并阻碍 EMT 程序。
采用高效液相色谱法（HPLC）对 MSD 提取物的成分进行表征。BALB/c-nu 小鼠被移植到 EGFP 标记的 SW480 CRC 细胞中，在接受不同剂量 MSD 治疗前后记录肿瘤重量和体积。采用酶联免疫吸附试验（ELISA）定量 TGF-β1 浓度。为了描绘 MSD 调节 EMT 和 CSC 之间的逻辑关系，在荷瘤小鼠和不同 CRC 细胞系中采用了 TGF-β/Smad 抑制剂和激活剂。采用流式细胞术分析癌症干细胞标志物。采用 CCK-8、划痕愈合和侵袭试验检测细胞迁移和活力的体外分析。采用免疫组织化学（IHC）和蛋白质印迹（WB）检测蛋白表达。采用 GraphPad Prism 8.0 对收集到的结果进行统计学分析。
MSD 治疗显著减小了结直肠癌肿瘤的大小并降低了小鼠血清中 TGF-β1 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并抑制了 CD133 干细胞。TGF-β/Smad 抑制剂通过去磷酸化 SMAD2/3 使 EMT 信号中和并降低多能性。同样，MSD 通过限制 TGF-β/Smad 信号诱导的 EMT 在体内减弱了多能性。MSD 抑制结直肠癌细胞增殖、迁移和侵袭。
MSD 抑制结直肠癌的生长。它通过抑制 TGF-β 诱导的 EMT 程序来抑制 CSC 的多能性。
