Modified Shenlingbaizhu Decoction represses the pluripotency of colorectal cancer stem cells by inhibiting TGF-β mediated EMT program

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.

Automated summary

The Modified Shenlingbaizhu Decoction (MSD) has been found to inhibit the pluripotency of colorectal cancer stem cells (CSCs) and impede the epithelial-mesenchymal transition (EMT) program. MSD treatment significantly reduces tumor size and TGF-beta 1 content in mice, while also decreasing the expression of pluripotent factors and stem cells in tumor tissues.