Effect of Baicalin on the Necroptosis of Mouse Colon Cancer in Vitro / 医药导报

ABSTRACT

Objective To investigate the effect of baicalin on CT26.WT cells of colon cancer in mice, and to discuss the cell death form. Methods CT26.WT cells were divided into four groups including of control group , routine cultured in fresh medium, the baicalin group, added with concentration of 100 μmol·L-1 baicalin, the z-VAD-fmk group, was added with final concentration of 20 μmol·L-1 z-VAD-fmk, and the combination group, added final concentration of 20 μmol·L-1 z-VADfmk,1 h before adding 100 μmol·L-1 baicalin. Then the inhibitory effect of baicalin on cell proliferation and cell viability were detected by CCK-8 method. The changes of nucleus were detected by DAPI staining, the ultrastructure of cells was observed by TEM, and the effect of baicalin on the expression of RIP3 gene and protein in cells was detected by QPCR method and Western blotting. Results Compared with control group, the differences of baicalin group and combination group had statistically significance (P<0.05) . cell death rate for control group was (10.54±0.19) ％ ,for baicalin group was (34.93±0.16) ％ ,for z- VAD group was (11.23±0.59) ％, and combination group was (23.27±1.20) ％ (P<0.01) . Compared with the normal control group, baicalin group showed nuclear concentration and fragmentation. there was obvious nuclear fragmentation in the combination group against baicalin group. The results of electron microscopy showed that the cells of baicalin were necrotic, cell swelling, mitochondria swelling and contents leaking. Baicalin group significantly up - regulated RIP3 mRNA expression (P < 0. 01) and enhanced RIP3 protein expression (P < 0. 05) . Conclusion Baicalin induces the necrosis of ct26. WT cells, and can significantly increase the gene and protein expression of RIP3.