Inhibitory Effect of Centipede Scolopendra Extract on Gallbladder Carcinoma via Up-regulation of PUMA

Background: To observe the effect of centipede scolopendra extract on gallbladder carcinoma (GBC) and further investigate its underlying mechanism. Methods: : The GBC cell line GBC-SD was purchased and cultured. Small interfering RNA (siRNA) was constructed. The mRNA expression levels of PUMA were measured by reverse transcription-quantitative PCR (RT-qPCR) and protein expression levels of p53, PUMA, Bax and Bcl-2 were measured by western blotting respectively. Viability and proliferation were detected using MTT and colony formation assays respectively. The apoptosis rate was determined by Annexin-V-FITC/PI double staining. Results: : The GBC-SD cell line was successfully obtained and cultured. MTT and colony formation assays demonstrated that the viability and proliferation of GBC-SD cells could be markedly inhibited by centipede scolopendra extract in a concentration-dependent manner in the limited concentration range. Following pretreatment with centipede scolopendra extract, RT-qPCR demonstrated that the expression levels of PUMA were markedly increased in the GBC-SD cells, and western blotting showed that the high expression of PUMA was accompanied by upregulation of Bax and downregulation of p53 and Bcl-2 in the GBC-SD cells. However, this effect has proved hard to reproduce after PUMA-siRNA. Flow cytometry revealed that the apoptosis rate was 9.8±2.2%, 25.3±3.6%, 10.6±2.0%, and 13.5±2.4% in control group, centipede scolopendra extract group, PUMA-siRNA group, and centipede scolopendra extract combined with PUMA-siRNA group respectively. Conclusions: : Centipede scolopendra extract could induce the apoptosis of GBC-SD cells by promoting the PUMA-Bax signaling pathway. It could serve as a potential novel therapy for GBC in clinical practice.