Effect of mucin 16 on the malignant phenotype of gastric linitis plastica
Objective To investigate the expression of mucin16(MUC16)gene in gastric linitis plastica and the effects of MUC16 on the proliferation and invasion of gastric cancer cells.Methods In to-tal,13 cases of gastric linitis plastica tissue samples from January 2009 to December 2017 in Zhejiang Cancer Hospital were collected and the expression of MUC16 was detected by immunohistochemical stai-ning.Fluorescence quantitative polymerase chain reaction(PCR)and Western blotting were used to ana-lyze the expression of MUC16.Cells with high MUC16 expression were selected and lentivirus was used to construct stable cell lines with low MUC16 knockdown.The experiment was divided into MUC16 knock-down group(MUC16-KO group)and negative control group(NC group).Cell counting kit-8(CCK-8)and clonal formation assays were used to detect the proliferation ability of different cells,and scratch healing as-say and Transwell assay were used to detect the migration and invasion ability of different cells.Chi-square test was used for qualitative data,and independent sample t-test or One-way analysis of variance was used for quantitative data.Results The expression level of MUC16 protein in leather stomach tissue samples was significantly higher than that in common gastric cancer tissues(2.99±0.30 vs.1.07±0.48,t=5.942,P＜0.05).The mRNA expression of MUC16 in human gastric cancer cell lines KATO Ⅲ,NUGC4,HGC27 and AGS was higher than that of human normal gastric epithelial cell line GES1(9.93± 0.79,8.69±0.54,6.37±0.61,3.58±3.30 vs.1.00±0.10,t=19.368,24.373,15.045,13.848,P＜0.05).The mRNA expression levels of MUC16 in human diffuse gastric cancer cell lines KATO Ⅲ,NUGC4 and HGC27 were higher than those in human gastric adenocarcinoma cell line AGS(9.93±0.79,8.69±0.54,6.37±0.61 vs.3.58±0.30,t=12.959,14.331,7.082,P＜0.05).The expression lev-els of MUC16 in human gastric cancer cell lines KATO Ⅲ,NUGC4,HGC27 and AGS were higher than those in human normal gastric epithelial cell lines GES1(0.79±0.01,0.62±0.02,0.53±0.00 and 0.38±0.01 vs.0.08±0.00,t=122.969,46.765,202.339,36.109,P＜0.05).The expression lev-els of MUC16 protein in human diffuse gastric cancer cell lines KATO Ⅲ,NUGC4,HGC27 were higher than those in gastric adenocarcinoma cells AGS(0.79±0.01,0.62±0.02,0.53±0.00 vs.0.38± 0.01,t=50.215,18.590,23.986,P＜0.05).The absorbance of MUC16-KO group was lower than that of NC group at 96 h[AGS cell line(0.86±0.02 vs.1.21±0.42),t=12.973,P＜0.05;HGC27 cell line(0.45±0.30 vs.1.04±0.11),t=8.936,P＜0.05].The clonal formation ability of two cell lines in MUC16-KO group was significantly lower than that of control group[AGS(191.67±2.08 vs.243.67± 3.51),t=22.062,P＜0.05;HGC27(66.67±2.52 vs.140.33±20.50),t=6.177,P＜0.05].The scratch healing rate of MUC16-KO group was significantly lower than that of control group[AGS(6.49± 0.56)％vs.(31.19±3.90)％,t=10.856,P＜0.05;HGC27(0.32±2.00)％vs.(4.32±2.00)％,t=3.441,P＜0.05].The invasion ability of two cell lines in MUC16-KO group was significantly lower than that of control group[AGS(2 049.00±268.53 vs.3 670.33±290.24),t=7.102,P＜0.05;HGC27(2 498.33±1 175.35 vs.6 869.67±548.88),t=5.837,P＜0.05].Conclusion MUC16 is highly expressed in gastric linitis plastica,and the expression level is higher than that of common gastric cancer.The high expression of MUC16 in gastric cancer cell lines can promote the proliferation,migration and invasion of gastric cancer cells.
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