| Objective To investigate the effect of Go-Ichi-Ni-San1（GINS1） on glycolysis, metastasis and invasion of colorectal cancer（CRC） cells and its possible molecular mechanism. Methods The expression of GINS1 in CRC cell lines SW480, HCT116, Lovo, SW620, Caco-2 and NCM460 was detected by Western blot and Quantitative reverse transcription polymerase chain reaction（qRT-PCR）. The HCT116 cell line（HCT116-shGINS1 group） and its negative control cell line（HCT116-shGINS1-NC group）, the SW480 cell line（SW480-GINS1-OE group） and its negative control cell line（SW480-GINS1-OENC group） were constructed by lentiviral infection. The migration ability of cells was detected by scratch test, and the invasion ability of cells was detected by Transwell test. The phosphorylation level of phosphatidylinositol 3-kinase（PI3K）/protein kinase B（AKT）/mammalian target of rapamycin（mTOR） pathway and the expression of glucose transporter type 1（GLUT1）, pyruvate kinase isozyme type M2（PKM2）, lactate dehydrogenase（LDHA） and hexokinase2（HK2） were detected by Western blot. PI3K activator 740 Y-P or inhibitor LY294002 were added to each group of cells, and the above indexes were detected again. Twenty-four nude mice were randomly divided into three groups: GINS1 overexpression group, GINS1 overexpression control group, GINS1 knockdown group and GINS1 knockdown control group, six in each group, the tumor formation and growth of nude mice in each group were observed. Results Compared with NCM460 cells, GINS1 was highly expressed in CRC cell lines. Compared with the HCT116-shGINS1-NC group, the cells in the HCT116-shGINS1 group showed a significant decrease in the ability of metastasis and invasion, the phosphorylation level of PI3K/AKT/mTORC1 pathway, and the expression level of glycolysis-related proteins（P＜0.05）. Compared with the SW480-GINS1-OENC group, in SW480-GINS1-OE group, the ability of metastasis and invasion was significantly enhanced, and the phosphorylation level of PI3K/AKT/mTORC1 pathway and the expression level of glycolysis-related proteins significantly increased（P＜0.05）. After addition of 740 Y-P, cells in the HCT116-shGINS1 group showed increased metastatic and invasive ability, increased phosphorylation of PI3K/AKT/mTORC1 pathway and increased expression of glycolysis-related proteins（P＜0.05）. After the addition of LY294002, cells in the SW480-GINS1-OE group showed decreased metastatic and invasive capacity, decreased phosphorylation levels of the PI3K/AKT/mTORC1 pathway, and decreased expression levels of glycolysis-related proteins（P＜0.05）. In vivo experiments showed that GINS1 overexpression promoted the growth of CRC, while GINS1 knockdown inhibited the growth of CRC（P＜0.05）. Conclusions GINS1 is highly expressed in CRC cells, and targeted activation of the PI3K/AKT/mTORC1 pathway enhances glycolytic processes and promotes tumor metastasis and invasion. |
