| PART Ⅰ:The expression and clinical significance of GALNT9 in gastric cancerObjective To explore the clinical significance of GALNT9 in gastric cancer by studying the expression of GALNT9 in gastric cancer tissues/paracancer normal tissues and gastric cancer cell lines.Method Immunohistochemistry was utilized to detect the expression of GALNT9 in gastric cancer/paracancer normal tissues and gastric cancer cell lines.Result The positive rate of GALNT9 expression in gastric carcinoma paracancer normal tissues was 86.3%,which was significantly higher than that in cancer tissues(59.1%), showing low statistically significant difference(P=0.1425).The positive rate of GALNT9 expression was correlated with lymph node metastasis(N stage).GALNT9 was relatively highly expressed in the gastric cancer cell line MGC803 and highly low in 7901.Conclusion GALNT9 is highly expressed in some gastric cancer tissues and positively correlated with lymph node metastasis of gastric cancer,suggesting that gastric cancer may participate in malignant biological behaviors such as invasion and metastasis of gastric cancer.The gastric cancer cell line MGC803 and 7901 can be used for further research.PART Ⅱ:The effects of the regulation of GALNT9 expression on the biological behavior of gastric cancer cellsObjective By silencing GALNT9 expression in gastric cancer cell MGC803,the biological behavior changes of gastric cancer cells after silencing were observed.The expression of GALNT9 gene in gastric cancer cells 7901 was overexpressed,and the biological behavior changes of gastric cancer cells were observed after overexpression.Method SiGALNT9 and negative siRNA were designed and transfected into gastric cancer MGC803 cell line with liposome reagent.Changes in protein expression level of gastric cancer MGC803 cells after transfection were detected by western-blot.The silencing expression of lentivirus Lv-GALNT9-RNAi and negative control virus CON077(hu6-mcs-ubiquitin-egfp-ires-puromycin)was designed,and the stable cell lines were obtained by assisting virus transfection in the biosafety cabinet of polybrene.The changes of GALNT9 protein expression,EMT-related proteins and signaling pathway Akt/m TOR/STAT3 in MGC803 cells and plasmid overexpression in 7901 after transfection were detected by western-blot.Transwell was used to detect changes in cell migration capacity.The changes of cell proliferation were detected by plate cloning.The changes of cell apoptosis were detected by flow cytometry.The tumorigenesis of nude mice was detected by subcutaneous tumorigenesis and tumor immunohistochemistry.Result SiGALNT9 can significantly silence the expression level of GALNT9 protein in MGC803 cells,and the migration and proliferation of MGC803 cells are enhanced,and the apoptosis is reduced.After successfully transfected with overexpressed plasmids,the expression level of GALNT9 protein in 7901 cells increased,and the migration and proliferation ability of the cells decreased,and the apoptosis increased.The expression levels of GALNT9 gene and EMT-related proteins Zo-1,E-cad and Vimentin in MGC803 were decreased,while the expression of GALNT9 gene in 7901 was increased.Silencing the GALNT9 expression in MGC803 can inhibit the Akt/m TOR/STAT3 signaling pathway,while the GALNT9 expression in over-expression of 7901 can activate the Akt/m TOR/STAT3 signaling pathway.After silencing MGC803 group with GALNT9,tumor formation of Lv-siGALNT9 group was larger than that of Con group,and immunohistochemistry of tumor specimens showed that Ki67 percentage of Lv-siGALNT9 group was significantly lower than that of Con group.Conclusion GALNT9 promotes EMT in gastric cancer cells and plays a positive regulatory role.GALNT9 plays an important regulatory role in the Akt/m TOR/STAT3 signaling pathway in gastric cancer cell lines.GALNT9 act S as an anticancer gene in gastric cancer cells. |
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