| Objective:To investigate the inhibitory effect,safety and possible mechanism of anlotinib hydrochloride combined with radiotherapy on human lung adenocarcinoma H1299 transplanted tumor in nude mice..Methods:Human lung adenocarcinoma H1299 cells were inoculated subcutaneously into the right axilla of nude mice.When the tumor volume was about 200 mm~3,the nude mice were randomly divided into 4 groups with 5 mice in each group.Divided into blank control group,anlotinib group(anlotinib 3mg/Kg gavage for two weeks),radiotherapy group(6Gy irradiation once),radiotherapy combined with anlotinib group(the same drug as anlotinib group,radiotherapy group)same as the radiotherapy group).The general state of the nude mice was observed.The transplanted tumor volume of the nude mice was measured every 3 days,and the growth curve of the tumor volume was drawn.After the treatment,the nude mice were sacrificed after anesthesia on the second day,the tumors were removed,and the tumor inhibition rate and combined effect of each group were calculated.Q value,apoptosis detection method to detect tumor cell apoptosis,hematoxylin-eosin staining method to observe the number of tumor cells in each group,the immunohistochemical method to detect Casapase3,c-Kit,CD34,Ki-67,Expression of HIF-1αand VEGF.CD34antibody detected tumor microvessel density(MVD)to determine tumor angiogenesis.The heart,liver,lung and kidney tissues of nude mice in each group were collected by HE staining to evaluate the damage of each organ.Results:After the treatment,the tumor growth of nude mice was inhibited in each treatment group,the activity status of nude mice in each group did not change significantly,and the tolerance among nude mice in each group was good during the experimental period.The tumor volume in the control group grew the most rapidly,and the difference was statistically significant(p<0.05).The anlotinib combined with the radiotherapy group had significantly more tumor-inhibiting effect than the anlotinib group and the radiotherapy group,and the difference was statistically significant(p<0.05).The tumor inhibition rates of the anlotinib group,the radiotherapy group,and the anlotinib combined radiotherapy group was53.25%,36.91%,and 71.19%,respectively.The Q value of the combined effect of anlotinib combined with the radiation therapy group was 1.01,suggesting that the combined regimen has a synergistic effect in the human lung adenocarcinoma H1299 nude mouse xenograft model.TUNEL test showed that the apoptosis of the anlotinib group and the anlotinib combined with the radiotherapy group increased,and the difference was statistically significant compared with the control group and the radiotherapy group(p<0.05);HE staining results showed that the tumor cells in the anlotinib combined with radiotherapy group’s tumor cell number decreased significantly;the immunohistochemical detection was determined by the IHC Profiler plug-in in Image J,and the expression of Casapase3 increased in the anlotinib group and the anlotinib combination group,and Ki-67,c-Kit,HIF-1α,CD34 were all decreased,and the difference was more significant in the anlotinib combined with radiotherapy group.The expression of VEGF in the anlotinib group and the anlotinib combined with the radiotherapy group was decreased,and the anlotinib combined with the radiotherapy group was more obvious,but the difference between the groups was not statistically significant.After treatment,there was no significant difference in HE staining of the heart,lung,kidney,and liver of nude mice in each group.Conclusion:1.The inhibitory effect of anlotinib is mainly through normalizing tumor blood vessels,improving hypoxia,inhibiting c-Kit to increase apoptosis,etc.It has a tumor-inhibiting effect on human lung adenocarcinoma H1299xenografts,and it has synergistic effect when combined with radiotherapy.2.In this study,the damage of each organ of anlotinib in the human lung adenocarcinoma H1299 xenograft model were tolerable whether it was a monotherapy or combined with radiotherapy. |
PDF Full Text Request