Deoxycholic Acid Induced The Expression Of MicroRNA223-3p In The Process Of Colorectal Cancer Incidence

Objective:?To observe the effect of deoxycholic acid promoting the proliferation of colonic epithelial cells?Exploring miRNA223-3p expression in colorectal cancer cells upregulated or down and whether miRNA223-3p can be used as an indicator of colorectal cancer diagnosis.Methods:?CCD841 cells were treated with different concentrations of deoxycholic acid(O?mol/L,5?mol/L,20?mol/L,50?mol/L,100?mol/L,150?mol/L,200?mol/L and 300?mol/L,respectively)after 24h,48h,72h,96h,the absorbance value(A)was determined by CCK8 method and the cell proliferative activity was measured.?The gene chip was used to detect the miRNA(three in experimental group and three in control group)of CCD841 cells treated with deoxycholic acid and the highest concentration of cell proliferation in this experiment,and this experiment was predicted MiRNA.?qRT-PCR detection of HT29 cells and CCD841 cells predicted miRNA223-3p expression is consistent;in colorectal cancer tissue and normal tissue to verify whether miRNA223-3p expression meets the requirementS?Results:?After adding different concentrations of deoxycholic acid to treat cells,CCK8 assay was used to detect the cell proliferation activity.It was found that deoxycholic acid could promote the proliferation of colorectal epithelial cell line,and the effect of promoting proliferation was dose-dependent.The corresponding cell proliferation rates were 131.3%,150.3%,122.0%,111.0%,119.0%,117.5%and 105.3%respectively at the concentrations of 0,5,20,50,100,150,200 and 300?mol/L after 24h treatment with deoxycholic acid.After 48h treatment with deoxycholic acid,the cell proliferation rates were 128.7%,144.8%,126.0%,117.0%,105.3%,90.9%and 81.4%,respectively.After 72 hours of deoxycholate treatment,The cell proliferation rate was 131.6%,111.6%,92.0%,99.2%,49.1%,42.4%,and 41.8%,respectively,besides,after 96 hours of deoxycholate treatment,cell proliferation rates were 131.2%,149.3%,114.4%,101.5%,90.1%33.3%and 20.9%,respectively.When the concentration was more than 200?mol/L,the activity of deoxycholic acid was toxic to colorectal epithelial cells at the concentration of 20?mol/L for 48h,and the cell proliferation was promoted when the concentration was 20?mol/L.?The strongest Gene microarray results showed that 56 miRNAs were differentially expressed,of which 25 were up-regulated and 31 were down-regulated.A total of 569 miRNAs with no significant difference were identified.A randomly selected down-regulated miRNA(miRNA223-3p)qRT-PCR experiments.?qRT-PCR detection miRNA223-3p lysis curve and amplification curve between HT29 cells and CCD841 cells were good,and the expression was lower in the experimental group than in the normal group.The miRNA223-3p expression was verified in randomly selected 12 pairs of colorectal cancer tissues and normal colorectal cancer tissues,which was consistent with the result of gene chip test.Conclude:?Deoxycholic acid can promote colorectal epithelial cell proliferation.And this promotion and deoxycholic acid concentration and duration of action,deoxycholic acid on the role of colorectal epithelial cells showed a two-way.When the concentration is less than 200umol/L,the effect of time does not exceed 24 hours can significantly promote the proliferation of CCD841.The results showed that when the concentration of deoxycholic acid was 0,5 and 20?mol/L,the effect of deoxycholic acid on the proliferation of colorectal epithelial cells was enhanced.The effect of promoting the proliferation was obvious at 24 hours.The promotion of 50?mol/L The effect than 20?mol/low.When the concentration is greater than 200umol/L,the role of time more than 24 hours or even cytotoxicity.?miRNA223-3p was detected by cDNA microarray in doxy cholic acid treated CCD841 cells,the expression of miRNA223-3p was down-regulated by qRT-PCR,and the expression of miRNA223-3p in the cells was both lower than the normal control group.To verify the miRNA22-3P expression in tissue,which is consistent with the results of microarray analysis.The expression of miRNA223-3p is not related to the patient's sex,age and tumor site,and is related to the TNM stage of the tumor.It is predicted that miRNA223-3p can be used as a new colorectal cancer detection way.