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目的:研究携带细胞穿膜肽11R和P53的溶瘤腺病毒SG7605-11R-P53对肝癌细胞的体外杀伤作用。方法:以本课题组前期实验构建的携细胞穿膜肽11R和P53的溶瘤腺病毒SG7605-11R-P53和不携11R的溶瘤腺病毒SG7605-P53感染肝癌细胞HepG2、SMMC-7721、Hep3B、Huh7和正常成纤维细胞株BJ,Western blotting检测感染后细胞P53和11R-P53的表达情况,TCID50法检测SG7605-11R-P53和SG7605-P53在肝癌细胞中的增殖能力,MTT法检测SG7605-11R-P53对肝癌细胞及正常细胞的杀伤作用。结果:SG7605-11R-P53和SG7605-P53能在肝癌细胞中高表达P53和11R-P53蛋白。SG7605-11R-P53可在HepG2、SMMC-7721、Hep3B和Huh7细胞中大量增殖,其增殖倍数是SG7605-P53的10~100倍,但在正常BJ细胞内几乎不增殖。SG7605-11R-P53在MOI=0.1时对Hep3B细胞的杀伤率达90%,对于正常BJ细胞只有当MOI=50时才有很弱的抑制作用;SG7605-11R-P53对4种肝癌细胞杀伤作用的大小依次为Hep3B、HepG2、Huh7和SMMC-7721细胞。结论:携带细胞穿膜肽11R和P53的SG7605-11R-P53溶瘤腺病毒体外对4种肝癌细胞株均有较好的靶向杀伤作用,尤其对Hep3B细胞的杀伤作用最强。
Objective: To study the in vitro cytotoxicity of oncolytic adenovirus SG7605-11R-P53 carrying cell penetrating peptide 11R and P53 on hepatoma cells. Methods: HepG2, SMMC-7721 and Hep3B cells were infected with oncolytic adenovirus SG7605-11R-P53 and oncolytic adenovirus SG7605-P53 without cell-11R and cell penetrating peptides 11R and P53 in our previous study , Huh7 and normal fibroblast cell lines BJ and Western blotting were used to detect the expression of P53 and 11R-P53. The proliferation of SG7605-11R-P53 and SG7605-P53 were detected by TCID50 method in vitro. MTT assay was used to detect the expression of SG7605- 11R-P53 on hepatoma cells and normal cells. Results: SG7605-11R-P53 and SG7605-P53 could highly express P53 and 11R-P53 in hepatoma cells. SG7605-11R-P53 can proliferate in HepG2, SMMC-7721, Hep3B and Huh7 cells in multiples of 10 to 100 times that of SG7605-P53, but hardly proliferate in normal BJ cells. SG7605-11R-P53 killing rate of Hep3B cells at MOI = 0.1 was 90%, and only weak inhibition was found at normal MOI = 0.1 for MOI = 50; SG7605-11R-P53 could kill 4 kinds of hepatoma cells Followed by Hep3B, HepG2, Huh7 and SMMC-7721 cells. CONCLUSION: SG7605-11R-P53 oncolytic adenovirus carrying cell-penetrating peptides 11R and P53 has good cytotoxicity against all four hepatocellular carcinoma cell lines in vitro, especially in Hep3B cells.