| ObjectiveColon cancer is one of the most common malignancies worldwide,which causes a major population of cancer-related deaths in the world.In the past two decades,the prevalence and mortality of colon cancer has been increased at an alarming rate for dietary habits have been changed to high fat or low textile fiber.Colon cancer is the result of multiple deregulations of gene networks,but the molecular mechanism remains to be elucidated.Gelsolin,which play the role in cytoskeletal actin depolymerization and restructuring,and involved in cell proliferation,cell apoptosis,cell movement even a variety of physiological functions.Gelsolin can be divided into cytoplasm and secretion.The cytoplasm gelsolin is involved in a variety of tumor initiation and progression,and secretory gelsolin mainly exists in the blood.This study aimed to explore the role of secretory gelsolin in the development of colon cancer and the diagnostic value of gelsolinMethodsImmunohistochemical method was used to detect the expression and the cellular localization of gelsolin in 36 cases of colon cancer and adjacent tissues and the expression of gelsolin protein in tissues was determined by Western Blot.The levels of serum were detected by ELISA detection kit and the ROC curve was used to evaluate the diagnostic value.Then Western Blot method was used to detect the expression of gelsolin in the colon cancer cell lines,and select the highest expression cell line HT29 and hardly expression cell line Lovo as the experimental cell lines.After that,we collected the supernatant of HT29 cell,then filtration by ultrafiltration tube and Western Blot test was used to verify gelsolin expression.Then add the supernatant to the Lovo cells,and join the gelsolin neutralizing antibodies,observe the influence of secretory gelsolin about the growth of tumor cell proliferation,migration and invasion.We transfected specific knockdown gelsolin RNA sequence to HT29 cell,then verified the effectiveness by fluorescence quantitative PCR and Western Blot assays.We collected HT29 cell supernatant after the transfection,and filtration and verify then added into Lovo cells and researched the role of tumor cell migration and invasion.The human recombinant protein gelsolin was added to stimulate Lovo cells to observe its effect on tumor cell migration and invasion.Then the human recombinant protein gelsolin was used to stimulate Lovo cells for 48 hours,and the total protein was extracted,and the correlation between EMT and MMPs was detected by Western Blot assay.ResultsGelsolin expression was significantly reduced in colon cancer tissues compared with adjacent tissues,as well as in serum.Then we drew the receiver operating characteristic(ROC)curves,and the area under the curve(AUC)of gelsolin was 0.932(95%CI was 0.877-0.967).The AUC of CEA and CA199 was 0.751 and 0.638 respectively.The AUC of combined with gelsolin,CEA and CA199 was as high as 0.966.After real-time fluorescence quantitative PCR and Western Blot verification,the cell line HT29 with high expression gelsolin was selected,and Lovo,which was hardly expression gelsolin was selected as the experimental cell of this study.Collected HT29 cell supernatant and filtration by ultrafiltration filter tube then added into Lovo cell,with the blank control group and to join the supernatant and gelsolin neutralizing antibody group.We found that there was no any effect on cell proliferation,but it could restrain Lovo cell migration and invasion ability.After collecting and filtering the HT29 cells from gelsolin,it was found that gelsolin was able to promote the migration and invasion of cells compared with the untreated control group.The recombinant protein gelsolin was used to stimulate Lovo cells to inhibit the migration and invasion ability of Lovo cells,and the inhibition ability increased with the increase of gelsolin concentration.The mRNA levels and protein expression levels of mmp-2 and mmp-9 were inhibited after the addition of the recombinant protein gelsolin,and the EMT related molecules were not affected significantly.ConclusionGelsolin was significantly reduced in colon cancer tissues and decreased in serum of colon cancer patients.Gelsolin and CEA,CA199 combined with the area of 0.966,indicated that gelsolin may have a certain value in the diagnosis of colon cancer.Secretory gelsolin has no obvious effect on the proliferation and growth of tumor cells,but can inhibit cell migration and invasion ability,and down-regulate mmp-2,mmp-9 protein expression level. |
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