| Objective: 1.To investigate the effect of inhibiting vimentin(VIM)expression using RNA interference technology on cell division and cell cloning in hepatoportal cholangiocarcinoma cell line QBC939;2.To construct a nude mouse model of hilar cholangiocarcinoma and explore the effect of inhibiting the expression of VIM on the growth of subcutaneously transplanted tumor in nude mice;3.To preliminarily reveal the mechanism of vimentin in the development of hilar cholangiocarcinoma.Methods:1.Three groups of Small interfering RNA(siRNA)with different sequences of nucleic acids(siRNA-VIM1,siRNA-VIM2,siRNA-VIM3)and their negative control sequences(siRNA-NC)were designed and synthesized.2.The hilar cholangiocarcinoma cell line QBC939 were cultured in vitro.The experimental group(transfected with siRNA-VIM1,siRNA-VIM2,siRNA-VIM3)and the control group(transfected with siRNA-NC),observe the effect on the number of QBC939 cell clones and cell division after inhibiting the expression of VIM.3.Eighteen SPF female nude mice aged 4 to 6 weeks old with a weight of(13±2)g were adaptiveiy reared for 7 days,QBC939 cell suspension of0.2m L and cell number of 2×106 was inoculated at 0.3cm of the back of the armpit of the right forelimb of nude mice,and the tumor-like tumors could be palpated at the inoculation site in about a week.Modeling is successful.Drug intervention was given when the tumor body grew to about 0.5cm in diameter,and randomly divided into Control group(normal saline 0.2m L/mouse),siRNA-VIM group(siRNA-VIM2 0.2m L/mouse),siRNA-NC group(siRNA-NC 0.2m L/mouse).Intravenously injected once a day for 15 days.After 15 days,the nude mice were killed by decapitation,the tumor was taken,and the tumor volume of each group was measured.The model was verified by HE staining,and the expression of VIM in the tumor was detected by q RT-PCR and immunohistochemical staining.Results: 1.siRNA-VIM and siRNA-NC were successfully transfected into hilar cholangiocarcinoma cell line QBC939.Compared with the control group,after inhibiting the expression of VIM by three groups of Small interfering RNA with different sequences of nucleic acids could significantly reduce the number of clones and cell division of QBC939 cells.2.Comparing the tumor volume of nude mice bearing hilar cholangiocarcinoma in each group,It was found that the tumor volume of siRNA-VIM group was smaller than that of Control group(P<0.05)and siRNA-NC group(P<0.05).3.q RT-PCR showed that the expression of VIM in siRNA VIM group decreased significantly compared with Control group and siRNA NC group(P<0.05).4.After HE staining was used to verify the modeling of nude mice,immunohistochemistry staining was used to detect that the expression of VIM in the tumor bearing nude mice with hilar cholangiocarcinoma was significantly lower than that in the control group and siRNA-NC group(P<0.05).Conciusion: 1.Inhibition of VIM expression can reduce the cell cloning and cell division ability of QBCQ939 cell line.2.Inhibiting the expression of VIM in QBC939 cells can effectively inhibit the growth of hilar cholangiocarcinoma transplanted tumor in nude mice.3.Inhibition of VIM expression is one of the important mechanisms of inhibiting the progression of hilar cholangiocarcinoma,and VIM plays an important role in the progression of hilar cholangiocarcinoma.4.VIM is an important marker of EMT,which is closely related to the development of many tumors.Deeply studying the mechanism of VIM in the development of tumors is of great significance for targeted treatment of tumors.VIM is expected to become a new target of anti-tumor treatment. |
PDF Full Text Request