| Objective: To investigate the effect of Scutellaria barbata D.Don on the self-renewal and differentiation of HT-29-derived colon cancer stem cells(HT-29CSC),to elucidate the molecular mechanism of HT-29 CSC from Hedgehog signaling pathway,and to provide scientific for the treatment of colorectal neoplasms Experimental basis.Methods: HT-29 CSC was screened from HT-29 cells by magnetic sorting.Using CCK8 method to detect the changes of absorbance after treatment of HT-29 CSC with different doses of Scutellaria barbata D.Don,studying their effect on the proliferation of HT-29 CSC.Drug treatment to detect HT-29 CSC into the sphere ability.3D culture and detection of ATP content in each group to study the growth status of HT-29 CS.The effect of Scutellaria barbata D.Don on its differentiation was observed under inverted microscope.The effects of Scutellaria barbata D.Don colorectal CSC markers CD133,Lgr5,Hedgehog genes Ptch1,Gli1,Nanong,CK20,Ki67 and target gene c-Myc transcription were studied by qRT-PCR.Western Blot Study on the Effects of Scutellaria barbata and Scutellarin on the Expression of Lgr5,Hedgehog Gene Gli1 and c-Myc in Colorectal.Results: Scutellaria barbata D.Don can significantly inhibit the proliferation and sphere ability of HT-29 CSC,The growth of HT-29 CSC showed that there was a significant inhibitory effect on the growth of HT-29 CSC.Scutellaria barbata D.Don have a significant effect on the differentiation of HT-29 CSC.Therefore,Scutellaria barbata D.Don can not only down regulated the levels of CSC markers CD133,Lgr5,Hedgehog genes Ptch1,Gli1,Nanong,CK20,Ki67 and target gene c-Myc mRNA but also down regulated the levels of c-Myc,Gli1,Lgr5 protein expression in HT-29 CSC.Conclusion: Scutellaria barbata D.Don can significantly inhibit the self-renewal and differentiation of HT-29 CSC cells,The mechanism is to regulate the expression of Hedgehog signaling pathway. |
