To Investigate The Effects Of Blueberry Anthocyanins On The Proliferation,Apoptosis And Histone Acetylation Modification Of Human Colon Cancer Cell Line HT-29

Objective:To investigate the effects of blueberry anthocyanin on the proliferation and apoptosis of human colon cancer cell line HT-29,and to investigate the changes of histone acetylation sites of H3K9,H3K14,H3K18 and H3K27 after co-incubation with the optimal concentration of blueberry anthocyanin.Methods:Under the same conditions,use cell culture technology to culture human colon epithelial cell line HCo Epi C and human colon cancer cell line HT-29,and observe the cell morphology,growth and proliferation of the two;select the logarithmic growth phase human colonic epithelial cell line HCo Epi C and human colon cancer cell line HT-29 were used in the experiment.Co-incubate human colon cancer cell line HT-29 with different concentrations(0,50,100,150,200ug/m L)of blueberry anthocyanin solutions,and use Real Time Cellular Analysis(RTCA x CELLigence)dynamics Observe the proliferation of human colon cancer cell line HT-29 to find the optimal concentration and action time of blueberry anthocyanins for co-incubation of human colon cancer cell line HT-29;different concentrations of blueberry anthocyanins(0,50,100,150,200ug/m L)after co-incubation of human colon cancer cell line HT-29,immunofluorescence(Annexin V-FITC/PI double staining method)was used to observe the co-incubation of human colon with different concentrations of blueberry anthocyanins under a fluorescence microscope The fluorescence color of the cancer cell line HT-29 was changed,and the apoptosis of the HT-29 cell line was analyzed;the nucleoplasmic separation technology was used to extract the human colon epithelial cell line HCo Epi C and the human colon cancer cell line cultured under equal time and conditions.HT-29 and the optimal concentration of blueberry anthocyanins were co-incubated with the nucleoprotein of human colon cancer cell line HT-29.Western blot(WB)was used to detect histones H3K9,H3K14,H3K18,H3K27 in different cell lines The level of acetylation modification and comparative analysis.Results:Under normal growth conditions,the growth rate of human colon cancer cell line HT-29 was significantly higher than that of human colon epithelial cell line HCo Epi C.RTCA results showed that after co-incubation of human colon cancer cell line HT-29 with different concentrations of blueberry anthocyanins(50,100,150,200ug/m L),its proliferation was inhibited compared with the control group(0ug/m L)Among them,when 50ug/m L blueberry anthocyanins were used for 25.5 hours,the inhibition rate was close to more than 50%,while the other blueberry anthocyanin concentration groups also showed significantly greater than 50% inhibition rates at25.5 hours,and the difference was statistically significant.Significance(P <0.01);immunofluorescence experiment showed that compared with the control group(0ug/m L),with the increase of blueberry anthocyanin concentration,HT-29 cell nuclear red staining increased,indicating an apoptotic human colon cancer cell line HT-29 gradually increased,and cell apoptosis was positively correlated with the concentration of blueberry anthocyanins.Western Blot showed that under the same conditions,compared with the human colonic epithelial cell line HCo Epi C,the H3K9,H3K14,H3K18,and H3K27 acetylation modification levels of the human colon cancer cell line HT-29 were down-regulated,and the difference was statistically significant(P<0.01);Compared with human colon cancer cell line HT-29,after adding 50ug/m L blueberry anthocyanin to co-incubate human colon cancer cell line HT-29,the histone H3K9,H3K14,H3K18,and H3K27 acetylation modification levels were up-regulated,The difference is statistically significant(P<0.01).Conclusions:The effect of blueberry anthocyanins on human colon cancer cell line HT-29 can inhibit proliferation and promote its apoptosis,the effect of blueberry anthocyanins on human colon cancer cell line HT-29 can up-regulate histones(H3K9,H3K14,H3K18,H3K27),the level of acetylation modification may be related to cell proliferation inhibition and cell apoptosis.