The Relationship Between NF-κB And Epithelial-to-mesenchymal Transition Of Radiation-induced Human Esophageal Carcinoma Cells

Objective:-To=Explore=the=relationship=between=NF-κB=and=epithelial-to-mesenchymal transition of human esophageal carcinoma cells andprovide theoretical foundation for formulating reasonable and effectiveindividualized treatment plan of esophageal carcinoma.Methods: Using the method of gradient irradiation dose, to make thehuman esophageal carcinoma epithelial cell Eca109to Eca109R60by thecumulative dose of60Gy. Changes in cell morphology were observed byoptical microscope. Meanwhile, real-time PCR and immunocytochemistrywere used to detect the mRNA and protein level of NF-κB, E-cadherin andVimentin in esophageal epithelial cell lines Eca109and Eca109R60.Result:1Observed under a microscope, compared with esophageal carcinomaepithelium Eca109, Eca109R60cell showed more irregular, elongatedspindle-shaped, loss of polarity, and more loose in cell adhesion.2The relationship between mRNA of NF-κB, E-cadherin and Vimentin.In non-irradiated Eca109cells, E-cad mRNA expression wassignificantly higher than that in Eca109R60cells（P=0.003）. The differencewas statistically significant（P＜0.01）. In non-irradiated cells Eca109, VimmRNA expression is significantly lower than that in irradiated cellsEca109R60(P=0.004). The difference is statistically significan（tP＜0.01）. Innon-irradiated cells Eca109, NF-κB mRNA expression is lower than that inirradiated cells Eca109R60(P=0.004). The difference is statistically significant（P＜0.01）. Relationship with each other after radiotherapy, NF-κB andVimentin, E-cadherin mRNA expression as follows. Linear relationshipbetween NF-κB and Vimentin is not statistically significant(P=0.747).However, their have a positive correlation in some extent. Linear relationshipbetween NF-κB and E-cadherin is also not statistically significant(P=0.061).It means NF-κB and E-cadherin expression have a negative correlation.3The relationship between the protein expression of NF-κB, E-cadherinand Vimentin. Results of immunocytochemistry showed Expression of theepithelial cell marker E-cadherin was mainly located in the cytoplasm,brownish yellow in esophageal carcinoma cells. The expression of E-cadherinprotein in cells Eca109was stronger than that in Eca109R60by microscopicobservation. Expression of the mesenchymal cell marker Vimentin proteinwas mainly located in the cytoplasm, brownish yellow in esophagealcarcinoma cells. The expression of Vimentin protein in cells Eca109waslower than that in Eca109R60by microscopic observation. The mostcommon subunits of NF-κB was p65/p65. NF-κB p65protein was expressedin the cytoplasm and nucleus in esophageal cancer cells. But the NF-κB p65protein was main expression in the nucleus when it was activated,brownish-yellow granules. The expression of NF-κB p65protein in cellsEca109was weaker than that in Eca109R60by microscopic observation.Conclusion:1After irradiation, the esophageal carcinoma cell Eca109R60showedradiation resistant.2The esophageal carcinoma radiation-resistant cell lines Eca109R60occured epithelial-to-mesenchymal transition，the marker downregulation ofE-cad, Vim upregulated.3In some extent, the expression of E-cad and NF-κB are negativelycorrelated. In a other hand, the expression of Vim and NF-κB are positivelycorrelated. Suggesting that NF-κB might be involved in esophageal carcinomacells occur epithelial-to-mesenchymal transition process, affecting theradiosensitivity of esophageal carcinoma.