| Objective Primary hepatocellular carcinoma(HCC)is a neoplastic disease with a very high degree of malignancy and a very poor prognosis,which has surpassed gastric cancer and ranks second in the cause of death of malignant tumors.Studies have shown that SAC3D1 is involved in the progression of gastric cancer,colon cancer,breast cancer and other tumors,which is closely related to its poor prognosis,but no studies have confirmed its role in HCC.In this study,the expression verification,clinical correlation analysis and cytology experiments of human HCC tissues were mainly used to verify the progression of SAC3D1 involved in tumors,and provide relevant theoretical basis for the treatment of HCC.Methods 1.The relationship between SAC3D1 gene and clinicopathological features of HCC patients was extracted based on online TCGA database.2.Collect tissue samples from HCC patients who underwent surgical resection in Gansu Provincial People’s Hospital from October 2020 to December 2022,and preliminary IHC experiments were used to detect whether SAC3D1 was expressed in liver cancer.RT-PCR experiments were used to verify the expression of SAC3D1 in cancer tissues and corresponding adjacent tissues,and secondly,the expression of SAC3D1 in the normal cell line LO2 and the hepatoma cell lines Hep G2,SMMC-7721,Huh7 and MHCC-97 cell lines were detected by RT-PCR and WB experiments,and the cell lines with the highest and lowest expression of SAC3D1 were selected from the hepatoma cell lines for subsequent experiments.3.The selected hepatoma cell lines were transfected by liposomes,the cell lines with the lowest expression of SAC3D1 were overexpressed,the cell lines with the highest expression were silenced,the transfection efficiency was observed under fluorescence microscopy,and RT-PCR and WB experiments were used to further verify the transfection.4.After the transfection was successful,the changes of cell proliferation ability,invasion ability and migration ability before and after transfection were detected by CCK-8 experiment,Transwell experiment and cell scratch experiment.5.Use statistical software to conduct statistical analysis.Results 1.Compared with adjacent tissues and normal hepatocyte lines,SAC3D1 was significantly more expressed in human tissues and cancer cell lines of liver cancer.Clinical data analysis showed that SAC3D1 gene expression was correlated with Vivacell,Age,BMI,Histologic grade,AFP values;(P<0.05).2.The clinical tissue specimen IHC showed that SAC3D1 was expressed in liver cancer tissue;The results of RT-PCR experiments showed that the expression of SAC3D1 in cancer tissues was significantly higher than that in the corresponding adjacent tissues.Through RT-PCR and WB experiments on cell lines,the results showed that the expression of SAC3D1 in hepatoma cell lines was higher than that of normal hepatic cell lines,among which the expression of Hep G2 was the highest and the expression of Huh7 was the lowest,so these two cell lines were selected for subsequent experiments.3.Huh7 cells were overexpressed by liposome transfection cell lines,and si RNA was used to interfere with the expression of SAC3D1 in Hep G2 cell lines,and the PCR and WB results showed that the difference was statistically significant(P<0.05),indicating that the transfection was successful.4.After overexpression treatment of Huh7 cells,the results of cell function experiments showed that the cell proliferation,invasion and migration ability were enhanced;Conversely,after interfering with the expression of Hep G2 cells,the proliferation,invasion and migration ability of cells was also weakened,and the difference was statistically significant(P<0.05).Conclusion 1.Compared with the normal liver tissues and cells,SAC3D1 were significantly lowly expressed in HCC tissues and cell lines.In addition,SAC3D1 expression was correlated with Vivacell,Age,BMI,Histologic grade,AFP,Child-Pugh grade values;2.SAC3D1 is involved in the progression of hepatocellular carcinoma and affects the prognosis of hepatocellular carcinoma. |
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