| Objective: Invasion and metastasis are important markers of cancer and the main causes of death in cancer patients.Therefore,the mechanism of lung cancer occurrence,metastasis and corresponding treatment research is an important direction of modern cancer research.Numerous studies have shown that epithelial-mesenchymal transition(EMT)plays an important role in the invasion and metastasis of tumors.Fucoxanthin,a carotenoid found in seaweeds,has been proved to have anti-tumor effects.The subjects of this study were human lung adenocarcinoma cells,lung adenocarcinoma and paracancerous tissues.The role of fucoxanthin in the proliferation,epithelial-mesenchymal transition and migration of human lung adenocarcinoma cells was studied in order to clarify the effect of fucoxanthin on the occurrence and development of lung cancer cells,and to further study the mechanism of fucoxanthin action,hoping to provide reference for the treatment of lung cancer.Methods:1.Selection of human lung adenocarcinoma cells: A549 cell line.2.CCK-8 method was used to observe the effect of fucoxanthin on proliferation inhibition of lung cancer A549 cells.3.TGF-?1 was used to induce EMT in lung cancer A549 cells.Western blotting assay was used to detect the expression of EMT marker proteins E-cadherin,Vimentin and Wnt/?-catenin pathway marker protein in lung cancer A549 cells under the intervention of TGF-?1 and/or fucoxanthin.4.Scratch test was used to detect the migration ability of lung cancer A549 cells under the intervention of TGF-?1 and/or fucoxanthin.5.Collection of human lung cancer tissues: The subjects were patients who underwent surgery in the Second Hospital Affiliated to Dalian Medical University from October 2018 to January 2019,and pathologically confirmed lung adenocarcinoma.A total of 38 cases of lung cancer and matched para-cancerous tissue specimens were collected.The average age of the patients was 61.36 ± 8.93 years old.There were 16 male patients and 22 female patients.The clinical stages were stage I-II.Among them,4 cases were poorly differentiated and 34 cases were moderately or highly differentiated.The number of cases with tissue size < 3cm was 30,and the number of cases with tissue size ? 3cm was 8.6.Immunohistochemical method was used to detect the expression of Wnt/?-catenin pathway protein ?-catenin in 38 pairs of lung adenocarcinoma tissues and adjacent tissues.Result: 1.CCK-8 results showed that fucoxanthin inhibited the proliferation of lung cancer A549 cells.And it is time and concentration dependent in a certain range.2.Western Blotting results showed that the expression of E-cadherin protein was decreased and Vimentin protein was increased when TGF-?1 acted on A549 cells.Meanwhile,the expression of ?-catenin,a marker protein of Wnt/?-catenin pathway,was increased(p<0.05).Compared with TGF-?1 alone,the expression of E-cadherin protein was increased,the expression of Vimentin and ?-catenin protein was decreased when TGF-?1 and fucoxanthin acted together on A549 cells.There were differences between the two groups(p<0.05).3.Scratch test showed that TGF-?1 significantly enhanced the migration ability of A549 cells,while fucoxanthin combined with TGF-?1 inhibited the migration ability,the difference was statistically significant(p<0.05).4.Immunohistochemical results showed that the high expression rate of ?-catenin in cancer tissues was higher than that in adjacent tissues(p<0.05).In cancer tissues,?-catenin was mainly abnormally expressed in cytoplasm and/or nucleus,while in para-cancerous tissues,the expression of ?-catenin was mainly in cell membrane(p<0.05).The results suggested that the expression of ?-catenin was not related to sex,age,tumor size and differentiation(p>0.05).Conclusion: Fucoxanthin can inhibit EMT,migration and cell proliferation of A549 cells.The mechanism may be through inhibiting Wnt/?-catenin pathway.At the same time,?-catenin abnormally expressed in early lung adenocarcinoma tissues. |
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