Neuritin Influenced TUBB3 Expression In Cells | Posted on:2020-11-23 | Degree:Master | Type:Thesis | Country:China | Candidate:M Huang | Full Text:PDF | GTID:2404330590981352 | Subject:Surgery | Abstract/Summary: | PDF Full Text Request | Objective:To investigate the effect of neuritin on the expression of β-tubulin TUBB3 protein at the cellular level,and to provide a theoretical basis for the study of paclitaxel chemoresistance.The aim is to provide a theoretical basis for the study of paclitaxel chemoresistance.Methods:The group was composed by pcDNA3.1-his-neuritin and pcDNA3.1-HA-TUBB3 of eukaryotic expression recombinant plasmids were co-transfected into 293 cell.The two kinds of protein in each group has been expressed in cell.Then,through the CO-IP experiment to verify the interaction between proteins.The pcDNA3.1-his-neuritin plasmid,pcDNA3.1-empty plasmid,pBS/U6-neuritin siRNA plasmid,pcDNA3.1-HA-TUBB3 plasmid were transfected into 293 T cells by liposome-mediated method.Neuritin has high expression,decreased expression and high expression of TUBB3.The expression of Neuritin protein and TUBB3 protein was detected by Western-blot technique.Results:1.The CO-IP assay was performed with his-tagged antibody and HA-tagged antibody,respectively.Western blotting confirmed that both the Neuritin and TUBB3 proteins could obtain the desired band at the desired position.2.After transfection of the corresponding plasmid,the expression of Neuritin protein and TUBB3 protein were detected by Western-blot technique.The expression of TUBB3 was highly expressed when Neuritin was highly expressed,and the expression of TUBB3 was decreased when Neuritin expression was decreased.The expression of TUBB3 protein was significantly different from that of the control group(P < 0.05).The high expression of TUBB3 had no effect on the expression level of Neuritin at the cellular level.The expression of Neuritin protein was not statistically different from the control group(P > 0.05).Conclusion:1.The results of CO-IP assay showed that Neuritin interacted with TUBB3 protein.2.Western-blot results indicated that Neuritin may act as a positive regulator or modifier of TUBB3 and regulate the expression of TUBB3. | Keywords/Search Tags: | Neuritin, TUBB3, Gastric Cancer, Western-blot, CO-IP | PDF Full Text Request | Related items |
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