| Background: In recent years,with the continuous advancement of colon cancer diagnosis and treatment technology,the overall survival rate of colon cancer patients has been significantly increased,but postoperative recurrence and metastasis,the overall treatment effect is not very satisfactory,targeting colon cancer cell specific changes.Finding specific targets is becoming the main direction of colon cancer treatment development.Objective: To investigate the molecular mechanism of leukemia inhibitory factor(LIF)regulating CD44 expression in colon cancer cells,and to evaluate the effect of LIF regulation mechanism on proliferation,drug resistance and tumorigenic ability of colon cancer cells,and to explore the role of LIF intervention in colon cancer,The significance of disease treatment.Methods: 1.Analysis of the relationship between LIF and colon cancer by bioinformatics data.2.MTT,Plate Cloning Assay and cell glomeration assay were used to detect the proliferation of colon cancer cells,the ability to pellet and the anti-LIF antibody to block the proliferation of colon cancer cells after the addition of exogenous LIF.3.The effect of LIF on the migration ability of colon cancer cells was observed by scratch test.The expression of marker m RNA of various stem cells in colon cancer cells after addition of exogenous LIF was detected by RT-PCR.4.Western Blot method was used to detect the activation of downstream cancer cell signaling pathway after addition of exogenous LIF.The expression of stem cell marker CD44,Nanog,OCT-4A protein and anti-LIF antibody blocked LIF on colon cancer cell signaling pathway STAT3 activation,activated the stem cell marker OCT-4A.5.Statistical analysis was performed on the data using SPSS 22.0,T-test analysis of variance were used to determine whether there was a significant difference.Results: Bioinformatic data analysis showed that LIF was significantly associated with colon cancer;The expression of LIF in colon cancer tissues was significantly higher than that in adjacent normal tissues,and the increased expression of LIF m RNA could significantly reduce the disease-free survival of patients.In the study of the effect of LIF on the phenotypic characteristics of colon cancer cell lines,the experimental results show that LIF can promote the proliferation,migration,colony formation and tumor ball formation ability of colon cancer cells;further screening markers related to colon cancer dryness,the results prove CD44 increased significantly after the addition of LIF;the results of bioinformatic data analysis showed that the expression of CD44 in colon cancer tissues was significantly higher than that in adjacent normal tissues.Combined with the analysis data of LIF,the correlation analysis between the two revealed that the expression of CD44 was elevated.High correlation was significantly associated with increased expression of LIF;these results indicate that LIF promotes the formation of colon cancer stem cells and is significantly associated with stem cell marker CD44;We further demonstrated by real-time PCR,western blot and FACS that the expression of CD44 was increased after exogenous addition of LIF and showed a time-dose dependence.In the study of the effect of LIF on the molecular mechanism of colon cancer cell lines,the experimental results show that LIF can activate JAK/STAT3 signaling pathway and activate stem cell marker OCT-4A,Nanog and SOX2.In order to study the significance of blocking LIF activity in vitro for the treatment of colon cancer,we prepared a neutralizing antibody that blocks the activity of LIF,an anti-LIF antibody,the experimental results show that anti-LIF antibody can inhibit the proliferation of colon cancer cells by LIF;block the activation of STAT3 signaling by LIF;can also block the activation of stem cell marker OCT-4A by LIF.Conclusion: 1.LIF promotes the proliferation of colon cancer cells.2.LIF can promote the expression of CD44 and OCT-4A proteins in colon cancer stem cells.3.Anti-LIF antibody can bind with LIF to block the effect of LIF on colon cancer cells. |
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