Objective:To investigate the mechanism of MiR-34a-mediated mTOR/4E-BP1 signaling pathway on apoptosis of esophageal cancer cells.Methods:Esophageal cancer cell lines were purchased from the ATCC cell bank and randomly...Objective:To investigate the mechanism of MiR-34a-mediated mTOR/4E-BP1 signaling pathway on apoptosis of esophageal cancer cells.Methods:Esophageal cancer cell lines were purchased from the ATCC cell bank and randomly divided into three groups. The first group, miR-34a group (miR-34a group): transfected miR-34a mimics into esophageal cancer cells by transfection;The second group, esophageal cancer group (EC group): simple esophageal cancer cell line without other treatment, normal culture;third group, idling group (ID group): transfected miR-34a negative control into esophageal cancer cell line . To explore MiR by qRT-PCR, TUNEL, cck-8 and Westernblot in the detection of miR-34a mRNA expression, apoptosis, proliferation, and protein content of mTOR/4E-BP1 in esophageal cancer cells. -34a mediates the mechanism of mTOR/4E-BP1 signaling pathway on apoptosis of esophageal cancer cells.Results:The results of qRT-PCR showed that the expression of miR-34a mRNA was the lowest in esophageal cancer cells in EC group, and the highest in miR-34a group in miR-34a group, miR-34a group and ID group, EC Compared with the group, the miR-34a mRNA content increased significantly, indicating successful transfection (allP<0.05). The results of TUNEL showed that the number of apoptosis of esophageal cancer cells in EC group was the least, and the apoptosis of esophageal carcinoma cells in miR-34a group was significantly increased. The apoptosis rate of miR-34a group was significantly higher than that of EC group and ID group (P<0.05). There was no significant difference in apoptosis between EC group and ID group. The OD value of cell proliferation in ID group and EC group was higher than that in miR-34a group, and there was significant difference between groups (P<0.05). The proliferation of esophageal cancer cells in EC group was the highest, and the number of esophageal cancer cells in miR-34a group was the least. There was no significant difference in the proliferation of esophageal cancer cells in the ID group compared with the EC group (P>0.05). The mTOR/4E-BP1 protein in esophageal cancer cells of EC group, ID group and miR-34a group was immunoblotted by Western blot. The gray scale showed that the mTOR/4E-BP1 protein content of EC group was the highest, EC group and Compared with the ID group, the protein content was similar, no significant (P>0.05), and the miR-34a group had the lowest protein content. Compared with the EC group and the ID group, the miR-34a group had lower protein content in esophageal cancer cells (allP< 0.05).Conclusions:MiR-34a may promote the apoptosis of esophageal cancer cells by inhibiting the expression of mTOR/4E-BP1 signaling pathway.展开更多
基金National Youth Science Fund Project(No.8180140119).
文摘Objective:To investigate the mechanism of MiR-34a-mediated mTOR/4E-BP1 signaling pathway on apoptosis of esophageal cancer cells.Methods:Esophageal cancer cell lines were purchased from the ATCC cell bank and randomly divided into three groups. The first group, miR-34a group (miR-34a group): transfected miR-34a mimics into esophageal cancer cells by transfection;The second group, esophageal cancer group (EC group): simple esophageal cancer cell line without other treatment, normal culture;third group, idling group (ID group): transfected miR-34a negative control into esophageal cancer cell line . To explore MiR by qRT-PCR, TUNEL, cck-8 and Westernblot in the detection of miR-34a mRNA expression, apoptosis, proliferation, and protein content of mTOR/4E-BP1 in esophageal cancer cells. -34a mediates the mechanism of mTOR/4E-BP1 signaling pathway on apoptosis of esophageal cancer cells.Results:The results of qRT-PCR showed that the expression of miR-34a mRNA was the lowest in esophageal cancer cells in EC group, and the highest in miR-34a group in miR-34a group, miR-34a group and ID group, EC Compared with the group, the miR-34a mRNA content increased significantly, indicating successful transfection (allP<0.05). The results of TUNEL showed that the number of apoptosis of esophageal cancer cells in EC group was the least, and the apoptosis of esophageal carcinoma cells in miR-34a group was significantly increased. The apoptosis rate of miR-34a group was significantly higher than that of EC group and ID group (P<0.05). There was no significant difference in apoptosis between EC group and ID group. The OD value of cell proliferation in ID group and EC group was higher than that in miR-34a group, and there was significant difference between groups (P<0.05). The proliferation of esophageal cancer cells in EC group was the highest, and the number of esophageal cancer cells in miR-34a group was the least. There was no significant difference in the proliferation of esophageal cancer cells in the ID group compared with the EC group (P>0.05). The mTOR/4E-BP1 protein in esophageal cancer cells of EC group, ID group and miR-34a group was immunoblotted by Western blot. The gray scale showed that the mTOR/4E-BP1 protein content of EC group was the highest, EC group and Compared with the ID group, the protein content was similar, no significant (P>0.05), and the miR-34a group had the lowest protein content. Compared with the EC group and the ID group, the miR-34a group had lower protein content in esophageal cancer cells (allP< 0.05).Conclusions:MiR-34a may promote the apoptosis of esophageal cancer cells by inhibiting the expression of mTOR/4E-BP1 signaling pathway.
