| Purpose: The effect of astemizole on the proliferation and cell cycle of colon cancer cells and the expression of Eag1 mRNA and protein in colon cancer cells were observed to clarify the mechanism of astemizole in inhibiting the development of colon cancer.Methods: The HT-29 cells were routinely cultured according to the ACTT method and randomly divided into 2 groups after passage 2-3 passages:1.No astemizole group 2.Astemizole intervention group,5uM astemizole 100 uL was added to the intervention group.The two groups were detected by EDU detection cell proliferation,using FCM to detect changes in cell cycle distribution;Using Q-PCR(quantitative Real-time)detection of Eag1 mRNA expression levels,WB(Western Blot)to detect the expression of Eag1 protein.Result: 1.Cell proliferation: EDU results showed that the cell proliferation rate of astemizole-treated group was lower than that of astemizole-free group(38.277±4.14,60.5733±6.632,P<0.05),and the difference was statistically significant.2.Cell cycle distribution: FCM assay Astemizole intervention group cells in the G0/G1 phase of the cell ratio was higher than that of the astemizole group(88.734±2.847,72.040±3.060,P<0.05),and resulted in S phase(5.293±0.378,7.780±1.008,P<0.05),G2/M phase reduction(4.010±0.026,5.637±0.351,P<0.05).Compared with astemizole-free group,astemizole can significantly block HT-29 cell cycle at G0/G1 phase,and the difference is statistically significant.3.Eag1 mRNA expression level: Q-PCR results showed that the relative expression of Eag1 mRNA in the astemizole-treated group was lower than that in the group without astemizole(1.000±0.056,3.240±0.207,P<0.05),and the difference was statistically significant.4.Eag1 protein expression: WB test results showed that the expression of Eag1 protein in the astemizole-treated group was lower than that in the group without astemizole(0.477±0.067,1.023±0.059,P<0.05),and the difference was statistically significant.Conclusion: 1.Astemizole may inhibit proliferation of colon cancer cells through Eag1,change the cell cycle distribution,and arrest cells in G0/G1 phase.2.Astemizole may inhibit the expression of Eag1 mRNA and protein in colon cancer cells. |
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