The Effects of Coix Seed Extract on the Proliferation, Invasion, Chemotherapy Sensitivity, and Shh/Gli1 Pathway of Colon Cancer Cells
CHEN Xiaoyan, JI Xiuhai*, YU Xiaoyu, LU Minjie, WANG Xuezhong
This study aims to investigate the effects of CSE (Coix seed extract) on the proliferation, invasion, chemotherapy sensitivity, and prognosis of colon cancer cells HCT116 based on the Shh/Gli1 pathway. HCT116 cells were cultured in vitro, and the CCK-8 (cell counting kit-8) was used to detect the proliferative activity of different concentrations of CSE on HCT116 cells. HCT116 cells were stochastically grouped into a control group (C group), an oxaliplatin group (Oxa group), a CSE group, a CSE+Oxa group, and a CSE+Oxa+PUR (purmorphamine) group. The plate cloning experiment was performed to detect cell proliferation ability. The scratch healing experiment was performed to measure cell migration ability. Transwell method was used to measure cell invasion ability. The one-step TUNEL in situ apoptosis detection kit was performed to measure cell apoptosis rate. Immunoblotting was used to measure the levels of Caspase-3, Bcl-2 (B-cell lymphoma-2), Bax (BCL2-associated X), Shh, and Gli1 proteins. This article established nude mouse xenograft tumor models to observe tumor growth. In vitro experiments showed that compared with the C group, the Oxa group and CSE group exhibited significantly decreased colony formation number, wound healing rate, cell invasion number, and protein expression levels of Bcl-2, Shh, and Gli1 (P<0.05), with significantly increased apoptosis rate and levels of Caspase-3 and Bax (P<0.05). Compared with the Oxa group or CSE group, the CSE+Oxa group showed remarkably reduced colony formation number, wound healing rate, cell invasion number, and protein expression levels of Bcl-2, Shh, and Gli1 (P<0.05), while the apoptosis rate and levels of Caspase-3 and Bax were significantly elevated (P<0.05). Additionally, com- pared with the CSE+Oxa group, the CSE+Oxa+PUR group displayed significantly increased colony formation number, wound healing rate, cell invasion number, and protein expression levels of Bcl-2, Shh, and Gli1 (P<0.05), with notably decreased apoptosis rate and levels of Caspase-3 and Bax (P<0.05). In vivo experiments demonstrated that compared with the C group, the Oxa group and CSE group had significantly reduced tumor weight, tumor volume (P<0.05), along with increased growth inhibition rate in a dose-dependent manner. Compared with the Oxa group or CSE group, the CSE+Oxa group exhibited further decreased tumor weight, tumor volume (P<0.05), and the growth inhibition rate was significantly enhanced (P<0.05). Collectively, CSE can inhibit the proliferation, mi- gration, and invasion of HCT116 cells and enhance their chemosensitivity, whose mechanism may be associated with the inhibition of the Shh/Gli1 signaling pathway.
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