Objective To investigate the effect and mechanism of G protein subunit beta 2(GNB2)on the migration and metastasis of colorectal cancer cells.Methods The human embryonic kidney cells 293FT in the logarithmic growth phase were randomly divided into the control group and the shGNB2 group.The 293 FT cells in the control group were transfected with PSPAX2,PMD2G,and Control plasmids,while the 293FT cells in the shGNB2 group were transfected with PSPAX2,PMD2G,and shGNB2 plasmids;and the viral supernatants were collected,respectively.Human colorectal cancer cells HCT116 and RKO in the logarithmic growth phase were divided into the control group and the shGNB2 group according to the random number table method.HCT116 and RKO cells in the control group were transfected with the viral supernatants from the control group of 293 FT cells;HCT116 and RKO cells in the shGNB2 group were transfected with the viral supernatants from the shGNB2 group of 293FT cells.The expression of GNB2 mRNA in HCT116 and RKO cells in the control group and the shGNB2 group was detected by real-time fluorescence quantitative polymerase chain reaction.The expressions of GNB2,Vimentin,N-cadherin and E-cadherin proteins in HCT116 and RKO cells in the control group and the shGNB2 group were detected by Western blot.The wound-healing rates of HCT116 and RKO cells in the control group and the shGNB2 group were measured by wound-healing assay.The migration of HCT116 and RKO cells in the control group and the shGNB2 group was detected by Transwell assay.A total of 2 x 106 HCT116 cells were taken from the control group and the shGNB2 group,respectively;and which were injected subcutaneously in nude mice,and the subcutaneous tumors were removed after 3 weeks and cut into small tissue pieces of 1 mm3.Eight female nude mice aged 4-5 weeks were divided into the control group and the shGNB2 group according to the random number table method,with 4 mice in each group.The tissue pieces of subcutaneous tumors from HCT116 cells in the control group were inoculated in the intestinal plasma membrane at the ileocecal junction of nude mice in the control group;the tissue pieces of subcutaneous tumors from HCT116 cells in the shGNB2 group were inoculated in the intestinal plasma membrane at the ileocecal junction of nude mice in the shGNB2 group;the death time of nude mice was recorded,and the liver was removed to observe the number of liver tumor metastasis.Results The relative expressions of GNB2 mRNA and protein in HCT116 and RKO cells in the shGNB2 group were significantly lower than those in the control group(P＜0.01).After 48-hour culture,the wound-healing rates of HCT116 and RKO cells in the shGNB2 group were significantly lower than those in the control group(P＜0.01);the numbers of the migrated HCT116 and RKO cells in the shGNB2 group were significantly less than those in the control group(P＜0.01).The number of intrahepatic metastatic tumors in nude mice in the shGNB2 group was significantly less than that in the control group(P＜0.05).The relative expressions of Vimentin and N-cadherin proteins in HCT116 and RKO cells in the shGNB2 group were significantly lower than those in the control group,while the relative expression of E-cadherin protein was significantly higher than that in the control group(P＜0.01).Conclusion Inhibiting the expression of GNB2 can effectively repress the metastatic ability of colorectal cancer cells.The mechanism may be related to GNB2's involvement in the metastasis of colorectal cancer cells by regulating the epithelial-mesenchymal transition process.
colorectal cancerG protein subunit beta 2migrationmetastasis
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