Ultrasound targeted microbubble destruction combined with oncolytic adenovirus therapy in treatment of pancreatic cancer in mice
Objective To explore the efficacy of ultrasound targeted microbubble destruction(UTMD)combined with oncolytic adenovirus therapy(oAd)for pancreatic cancer in mice.Methods Subcutaneous injection of Panc-02 cells(2 x 106 cells per mouse)was performed on the right forelimb of C57BL/6 mice.A total of 55 mouses with tumor volumes ranging from 100-150 mm3 were selected and randomly divided into 5 groups,with 11 mouses in each group.The grouping and intratumoral administration were as follows:NC group(phosphate solution 100 μL),UTMD group(microbubble solution 100 μL and ultrasonic microbubble destruction treatment for 5 min),oAd group(108 PFU/mL oncolytic adenovirus solution 100 μL),oAd+MBs group(108 PFU/mL oncolytic adenovirus microbubble mixture 100 μL),oAd+UTMD group(108 PFU/mL oncolytic adenovirus microbubble mixture 100 μL and ultrasound microbubble destruction treatment for 5 min).All groups were treated every alternative days for 5 times,and the tumor volumes were measured also every alternative days.After 24 h of the third administration,6 mouses in each group were randomly selected and sacrificed.The tumors were obtained and tumor tissue sections and cell suspensions were prepared.HE staining was used to observe the necrosis of tumor tissues,E1A antibody staining was used to observe the distribution of oAds in tumor tissues in each group,CD3 antibody staining was used to count the number of CD3+T cells in tumor tissues,Tunel staining and flow cytometry were used to analyze the apoptosis of tumor tissues.Results When the experiment was terminated at 14 days,the tumor volume growth in the oAd+UTMD group was significantly slower than that in the oAd group(P＜0.05).The E1A protein staining of oAds in the cytoplasm was the deepest in the oAd+UTMD group and the lightest in the oAd group.The ratios of necrotic area in the oAd+UTMD group and the oAd group were 9.50±0.60 and 3.51±0.24,respectively,which were significantly different from those in the NC group(P＜0.05).CD3+T cells were accumulated in the tumors of the three groups of mice treated with oAd.The number of CD3+T cells in the tumors of the oAd+UTMD group(196.33±12.58)was significantly higher than that of the oAd group(120.67± 12.90,P＜0.05).The total apoptosis rate and Tunel staining fluorescence range in the oAd+UTMD group were significantly higher than those in the oAd group.Conclusion UTMD can enhance the killing effect of oAd on pancreatic tumor,promote the transfection of oAd in tumor,assist the enrichment of CD3+T cells in tumor,and induce apoptosis and necrosis of tumor cells.
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维普
