Nitidine chloride inhibits the appearance of cancer stem-like properties and regulates potential the mitochondrial membrane alterations of colon cancer cells

Nitidine chloride (NC) is a natural alkaloid that can inhibit tumor growth and induce apoptosis in varieties of cancers. However, the effec12/268t of NC on colon cancer (CC) cells has not been extensively studied.

NC treatment inhibited the proliferation and stemness of CC tissues, promoted the apoptosis of tumor tissues, downregulated the expression of p-ERK and p-AKT in tumor tissues, which suggests that NC may play an important role in regulating ERK and AKT pathways.

Conlon cancer SW480 cells was treated with different concentrations of NC (0.25, 0.5, 1, 2.5, 5, 10, 25, 50, 100, and 200 µM) in DMEM medium for 24 hours. Western blotting (WB) was used to detect the expression of related proteins, such as Ki67, PCNA, NANOG, SOX2, OCT4, Bcl-2, Bax, Caspase-3, Caspase-9, ERK1/2, p-ERK1/2, AKT, p-AKT, STAT3, p-STAT3, P65 and p-P65. The pellet formation experiment was used to detect the pellet formation of stem cells. The JC-1 experiment was used to detect the change of mitochondrial membrane potential. Kit was performed to detect the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA). In vivo experiments were used to verify the

NC significantly inhibited the expression levels of Ki-67 and a proliferating cell nuclear antigen (PCNA). NC can reduce the pellet colony and pellet size of tumor stem cells and block the stem cell characteristics of CC cells. The corresponding stem cell marker molecules NANOG, SOX2, and OCT4 were also downregulated. NC treatment induced the mitochondrial membrane potential depolarization of CC cells. The expression of pro-apoptotic proteins such as caspase-3, caspase-9, and Bax were upregulated, while the expression level of apoptotic Bcl-2 was significantly down-regulated. Moreover, NC reduced SOD activity and MDA content in CC cells. In addition, studies on pathway phosphorylation have shown that NC inhibits the expression of p-erk and p-akt proteins. Finally, the results were further confirmed by experiments in nude mice. NC inhibited tumor growth in mice. NC promoted apoptosis in tissues. NC inhibited the expression of Ki67 and OCT4 in tissues. NC inhibited the phosphorylation of pathway proteins ERK1/2 and AKT in tissues. Conclusions: NC treatment inhibited the proliferation and stemness of CC tissues, promoted the apoptosis of tumor tissues, downregulated the expression of p-ERK and p-AKT in tumor tissues, which suggests that NC may play an important role in regulating ERK and AKT pathways.