Figure 4.

Systemic Administration of 18E7 gRNAs +WT Cas9 Packaged in Stealth Liposomes Effectively Eliminated HeLa Xenografts and Prolonged Survival

(A) HeLa cells were subcutaneously inoculated in Rag1 mice and allowed to grow to 50 mm³ before being treated with a total of 10 μg/dose of plasmid DNA coated in stealth liposomes (18E7 sgRNA⁺ WT Cas9 plasmids [treatment], nonspecific gRNA + WT Cas9 [control], or PBS [untreated]). Tumor volume was monitored for 46 days (experiment endpoint, tumor size = 1,000 mm³). The arrows represent the days of treatment injection. The legend illustrates whether 18E7 or control treatments were injected to treatment arms (#1 and #2). Data are presented as mean ± SD. (B) The survival analysis of established HeLa xenografts in Rag1 mice after 18E7 targeted treatment, control (nonspecific gRNA + WT Cas9), or untreated (PBS only) in days. (C) The apoptotic cell count in anti-caspase-3-stained tumor tissues in 18E7, control, and untreated tumor sections. (D) Immunohistochemical staining of tumor tissues with H&E or anti-cleaved caspase-3 antibody in 18E7 or control treated tumor specimens. (E) Immunohistochemical staining of tumor tissues after seven doses of treatment (treatment #2 group) with 18E7 sgRNA + WT Cas9, with H&E or p16 antibody. Statistical significance was assessed by ANOVA with post hoc analysis. *p < 0.05, **p < 0.01, ***p < 0.001.