| Objective:(1)To explore the effect of UBE2 S on the proliferation and mechanism of lung adenocarcinoma in vivo.(2)To explore the effect of Udabara-8 powdered medicine on the proliferation of lung adenocarcinoma.Method:1.Preparation of tumorigenic cells: A549 cells were cultured and subcultured,selected cells were infected with lentivirus,and the infection efficiency was observed after 72 hours.2.q PCR was used to detect m RNA expression after UBE2 S gene knock-down.3.CCK-8was used to detect the drug sensitivity IC50;for 72h;4.Modeling and drug administration:Randomly grouping according to body weight;NC groups were injected with A549 cells without UBE2 S gene knockout.KD groups were injected with A549 cells with UBE2 S gene knocked out.Luteolin 0.05g/kg,Udabara-8 powdered medicine 5g /kg and 10g/kg were given by gavage once a day for 28 days.The changes of tumor were observed,and the relevant data of tumor were recorded and compared.5.HE staining: The histopathological changes were observed.6.Immunohistochemical method: The expression of UBE2 S and P53 protein in cancer tissues was detected.Results(1)Preparation of tumor cells: After 72 hours of infection,the fluorescence rate of each group was above 80%,and the cell state was normal,so the preparation of tumor cells was successful.(2)QCPR detection: After infection with sh RNA lentivirus,the expression of UBE2 S gene in A549 cells in experimental group was down-regulated at m RNA level(p<0.05).(3)CCK-8 method: The proliferation of A549 cells was obviously inhibited by luteolin after 72 hours in each group,and the difference was statistically significant(P < 0.05).(4)The change of tumor volume: the tumor was formed after 18 days of modeling.Comparing the tumor volume at10 time points,compared with the NC group,the tumor volume in each NC administration group was significantly reduced,with statistical significance(P﹤0.05).Compared with KD group,the tumor volume in KD+ monomer group was significantly reduced,with statistical significance(P﹤0.05).Compared with KD group,the tumor volume in NC group was significantly reduced,and the difference was statistically significant(P ﹤ 0.05).(5)Changes of tumor weight:Compared with NC group,the tumor weight in each NC administration group was significantly reduced,with statistical significance(P﹤0.05).Compared with KD group,the tumor weight in KD+ monomer group was significantly reduced,with statistical significance(P﹤0.05).(6)HE staining method: In NC group,compared with each administration group,there were more cancer cell necrosis among tissues.Compared with KD group,there were more cancer cell necrosis in KD+DT group.Compared with KD+ compound low concentration group and KD+high concentration group,the cancer cell necrosis in KD group is relatively less.(7)Immunohistochemical method: The protein expression levels of UBE2 S and P53 in tumor tissues of NC group,KD group and KD+DT group decreased.The comparison between NC group and KD group showed that the results were statistically significant(P<0.05).Conclusion:1.Knocking down UBE2 S gene can inhibit the growth and proliferation of A549 cells in nude mice.2.Udabara-8 flavored powder can inhibit the proliferation of A549 cells.3.Udabara-8 powder and luteolin inhibit the proliferation of lung adenocarcinoma through UBE2 S. |
